Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 10

Search results for: Miroslava Simakova

10 Fatty Acid Translocase (Cd36), Energy Substrate Utilization, and Insulin Signaling in Brown Adipose Tissue in Spontaneously Hypertensive Rats

Authors: Michal Pravenec, Miroslava Simakova, Jan Silhavy


Brown adipose tissue (BAT) plays an important role in lipid and glucose metabolism in rodents and possibly also in humans. Recently, using systems genetics approach in the BAT from BXH/HXB recombinant inbred strains, derived from the SHR (spontaneously hypertensive rat) and BN (Brown Norway) progenitors, we identified Cd36 (fatty acid translocase) as the hub gene of co-expression module associated with BAT relative weight and function. An important aspect of BAT biology is to better understand the mechanisms regulating the uptake and utilization of fatty acids and glucose. Accordingly, BAT function in the SHR that harbors mutant nonfunctional Cd36 variant (hereafter referred to as SHR-Cd36⁻/⁻) was compared with SHR transgenic line expressing wild type Cd36 under control of a universal promoter (hereafter referred to as SHR-Cd36⁺/⁺). BAT was incubated in media containing insulin and 14C-U-glucose alone or 14C-U-glucose together with palmitate. Incorporation of glucose into BAT lipids was significantly higher in SHR-Cd36⁺/⁺ versus SHR-Cd36⁻/⁻ rats when incubation media contained glucose alone (SHR-Cd36⁻/⁻ 591 ± 75 vs. SHR-Cd36⁺/⁺ 1036 ± 135 nmol/gl./2h; P < 0.005). Adding palmitate into incubation media had no effect in SHR-Cd36⁻/⁻ rats but significantly reduced glucose incorporation into BAT lipids in SHR-Cd36⁺/⁺ (SHR-Cd36⁻/⁻ 543 ± 55 vs. SHR-Cd36⁺/⁺ 766 ± 75 nmol/gl./2h; P < 0.05 denotes significant Cd36 x palmitate interaction determined by two-way ANOVA). This Cd36-dependent reduced glucose uptake in SHR-Cd36⁺/⁺ BAT was likely secondary to increased palmitate incorporation and utilization due to the presence of wild type Cd36 fatty acid translocase in transgenic rats. This possibility is supported by increased incorporation of 14C-U-palmitate into BAT lipids in the presence of both palmitate and glucose in incubation media (palmitate alone: SHR-Cd36⁻/⁻ 870 ± 21 vs. SHR-Cd36⁺/⁺ 899 ± 42; glucose+palmitate: SHR-Cd36⁻/⁻ 899 ± 47 vs. SHR-Cd36⁺/⁺ 1460 ± 111 nmol/palm./2h; P < 0.05 denotes significant Cd36 x glucose interaction determined by two-way ANOVA). It is possible that addition of glucose into the incubation media increased palmitate incorporation into BAT lipids in SHR-Cd36⁺/⁺ rats because of glucose availability for glycerol phosphate production and increased triglyceride synthesis. These changes in glucose and palmitate incorporation into BAT lipids were associated with significant differential expression of Irs1, Irs2, Slc2a4 and Foxo1 genes involved in insulin signaling and glucose metabolism only in SHR-Cd36⁺/⁺ rats which suggests Cd36-dependent effects on insulin action. In conclusion, these results provide compelling evidence that Cd36 plays an important role in BAT insulin signaling and energy substrate utilization.

Keywords: brown adipose tissue, Cd36, energy substrate utilization, insulin signaling, spontaneously hypertensive rat

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9 Upcycling of Inorganic Waste: Lessons Learned and Outlook for the Future

Authors: Miroslava Hujová, Patricia Rabello Monich, Jozef Kraxner, Dusan Galusek, Enrico Bernardo


Inorganic waste upcycling offers a solution how to avoid landfilling and how to save raw materials at the same time. However, its practical implementations in Slovakia and elsewhere in Europe, are rather limited despite the potential smaller countries like Slovakia have their advantage in closely-knitted inorganic materials industry. One part of discussion should include an overview of wastes that can be possibly used for upcycling, i.e. fly ashes, red mud, glass cullets, vitrified bottom ashes etc. These wastes can be processed by a variety of strategies, the one of our choice, alkali activation, opens the possibility for the formation of novel materials at almost negligible energetic expense. In the research, these materials are characterized by comprehensive means (X-Ray Fluorescece, Diffraction methods, Thermal Analysis, Scanning Electron Microscopy, Mechanical tests and Chemical stability), which time and time again demonstrate their competitive properties against traditional materials available at the market. It is just a question for discussion why these materials do not receive more significant attention from industry and there is pressing interest for the solution of standing situation.

Keywords: upcycling, inorganic wastes, glass ceramics, alkali-activation

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8 Effect of Fly Ash Fineness on Sorption Properties of Geopolymers Based on Liquid Glass

Authors: Miroslava Zelinkova, Marcela Ondova


Fly ash (FA) thanks to the significant presence of SiO2 and Al2O3 as the main components is a potential raw material for geopolymers production. Mechanical activation is a method for improving FA reactivity and also the porosity of final mixture; those parameters can be analysed through sorption properties. They have direct impact on the durability of fly ash based geopolymer mortars. In the paper, effect of FA fineness on sorption properties of geopolymers based on sodium silicate, as well as relationship between fly ash fineness and apparent density, compressive and flexural strength of geopolymers are presented. The best results in the evaluated area reached the sample H1, which contents the highest portion of particle under 20μm (100% of GFA). The interdependence of individual tested properties was confirmed for geopolymer mixtures corresponding to those in the cement based mixtures: higher is portion of fine particles < 20μm, higher is strength, density and lower are sorption properties. The compressive strength as well as sorption parameters of the geopolymer can be reasonably controlled by grinding process and also ensured by the higher share of fine particle (to 20μm) in total mass of the material.

Keywords: alkali activation, geopolymers, fly ash, particle fineness

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7 Phthalate Exposure among Roma Population in Slovakia

Authors: Miroslava Šidlovská, Ida Petrovičová, Tomáš Pilka, Branislav Kolena


Phthalates are ubiquitous environmental pollutants well-known because of their endocrine disrupting activity in human organism. The aim of our study was, by biological monitoring, investigate exposure to phthalates of Roma ethnicity group i.e. children and adults from 5 families (n=29, average age 11.8 ± 7.6 years) living in western Slovakia. Additionally, we analysed some associations between anthropometric measures, questionnaire data i.e. socio-economic status, eating and drinking habits, practise of personal care products and household conditions in comparison with concentrations of phthalate metabolites. We used for analysis of urine samples high performance liquid chromatography and tandem mass spectrometry (HPLC-MS/MS) to determine concentrations of phthalate metabolites monoethyl phthalate (MEP), mono-n-butyl phthalate (MnBP), mono-iso-butyl phthalate (MiBP), mono(2-ethyl-5-hydroxyhexyl) phthalate (5OH-MEHP), mono(2-ethyl-5-oxohexyl) phthalate (5oxo-MEHP) and mono(2-etylhexyl) phthalate (MEHP). Our results indicate that ethnicity, lower socioeconomic status and different housing conditions in Roma population can affect urinary concentration of phthalate metabolites.

Keywords: biomonitoring, ethnicity, human exposure, phthalate metabolites

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6 Structural and Magnetic Properties of CoFe2O4:Nd3+/Dy3+/Pr3+/Gd3+ Nanoparticles Synthesized by Starch-Assisted Sol-Gel Auto-Combustion Method and Annealing Effect

Authors: Raghvendra Singh Yadav, Ivo Kuřitka, Jaromir Havlica, Zuzana Kozakova, Jiri Masilko, Lukas Kalina, Miroslava Hajdúchová, Vojtěch Enev, Jaromir Wasserbauer


In this work, we investigated the structural and magnetic properties of CoFe2O4:Nd3+/Dy3+/Pr3+/Gd3+ nanoparticles synthesized by starch-assisted sol-gel combustion method. X-ray diffraction pattern confirmed the formation of cubic spinel structure of rare-earth ions (Nd3+, Dy3+, Pr3+, Gd3+) doped CoFe2O4 spinel ferrite nanoparticles. Raman and Fourier Transform Infrared spectroscopy study also confirmed cubic spinel structure of rare-earth ions (Nd3+, Dy3+, Pr3+, Gd3+) substituted CoFe2O4 nanoparticles. The field emission scanning electron microscopy study revealed the effect of annealing temperature on size of rare-earth ions (Nd3+, Dy3+, Pr3+, Gd3+) substituted CoFe2O4 nanoparticles and particles were in the range of 10-100 nm. The magnetic properties of rare-earth ions (Nd3+, Dy3+, Pr3+, Gd3+) substituted CoFe2O4 nanoparticles were investigated by using vibrating sample magnetometer. The variation in saturation magnetization, coercivity and remanent magnetization with annealing temperature/ particle size of rare-earth ions (Nd3+, Dy3+, Pr3+, Gd3+) substituted CoFe2O4 nanoparticles was observed. Acknowledgment: This work was supported by the Ministry of Education, Youth and Sports of the Czech Republic – Program NPU I (LO1504).

Keywords: starch, sol-gel combustion method, rare-earth ions, spinel ferrite nanoparticles, magnetic properties

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5 Silica Nanofibres – Promising Material for Regenerative Medicine

Authors: Miroslava Rysová, Zdena Syrová, Tomáš Zajíc, Petr Exnar


Currently, attention of tissue engineers has been attracted to novel nanofibrous materials having advanced properties and ability to mimic extracellular matrix (ECM) by structure which makes them interesting candidates for application in regenerative medicine as scaffolding and/or drug delivering material. Throughout the last decade, more than 200 synthetic and natural polymers have been successfully electrospun leading to the formation of nanofibres with a wide range of chemical, mechanical and degradation properties. In this family, inorganic nanofibres represent very specific group offering an opportunity to manufacture inert to body, well degradable and in properties tunable material. Aim of this work, was to reveal unique properties of silica (SiO2, CAS 7631-86-9) nanofibres and their potential in field of regenerative medicine. Silica nanofibres were prepared by sol-gel method from tetraethyl orthosilicate (TEOS, CAS 78-10-4) as a precursor and subsequently manufactured by needleless electrospinning on NanospiderTM device. Silica nanofibres thermally stabilized under 200°C were confirmed to be fully biodegradable and soluble in several simulated body fluids. In vitro cytotoxicity tests of eluate (ES ISO 10993-5:1999) and in direct contact (ES ISO 10993-5:2009) showed no toxicity - e.g. cell viabilities reached values exceeding 80%. Those results were obtained equally from two different cell lines (Vero, 3T3). Non-toxicity of silaca nanofibres´ eluate was additionally confirmed in real time by testing on xCelligence (ACEA Biosciences, Inc.) device. Both cell types also showed good adhesion to material. To conclude, all mentioned results lead to resumption that silica nanofibres have a potential as material for regenerative medicine which opens door to further research.

Keywords: cytotoxicity, electrospinning, nanofibres, silica, tissue engineering

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4 Biological Control of Woolly Apple Aphid, Eriosoma Lanigerum (Hausmann) in the Nursery Production of Spruce

Authors: Snezana Rajkovic, Miroslava Markovic, Ljubinko Rakonjac, Aleksandar Lucic, Radoslav Rajkovic


Woolly apple aphid, Eriosoma lanigerum (Hausmann) is a widely distributed pest of apple trees, especially where its parasites have been killed by insecticides. It can also be found on pear, hawthorn, mountain ash, and elm trees. Relatively small to medium-sized aphids, characterized by a reddish-brown body, a blood-red stain when crushed and a fluffy, flocculent wax covering. Specialized dermal glands produce the characteristic fluffy or powdery wax, which gives E. lanigerum its characteristic 'woolly' appearance. Also, woolly apple aphid is a problemm in nursery production of spure.The experiments were carried out in the nursery “Nevade” in Gornji Milanovac, "Srbijasume" on the spruce seedlings, aged 2 years. In this study, organic insecticide King Bo, aqueous solution (a. i. oxymatrine 0.2% + psoralen 0.4%), manufacturer Beijing Kingbo Biotech Co. Ltd., Beijing, China. extracted from plants and used as pesticides in nursery production were investigated. King Bo, bioinsecticide is manufactured from refined natural herbal extract several wild medicinal plants, such as Sophora flavescens Ait, Veratrum nigrum L, A. Carmichael, etc. Oxymatrine 2.4 SL is a stomach poison that has antifeeding and repellent action. This substance stimulates development and growth in a host plant and also controls the appearance of downy mildew.The trials were set according to instructions of methods-monitoring of changes in the number of larvae and adults compared to before treatment. The treatment plan was made according to fully randomized block design. The experiment was conducted in four repetitions. The basic plot had the area of 25 m2. Phytotoxicity was estimated by PP methods 1/135 (2), the intensity of infection according to Towsend-Heuberger, the efficiency by Abbott, the analysis of variance with Ducan test and PP/181 (2).

Keywords: bioinsecticide, efficacy, nurssery production, woolly apple aphid

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3 Biofungicides in Nursery Production

Authors: Miroslava Markovic, Snezana Rajkovic, Ljubinko Rakonjac, Aleksandar Lucic


Oak powdery mildew is a serious problem on seedlings in nurseries as well as on naturally and artificially introduced progeny. The experiments were set on oak seedlings in two nurseries located in Central Serbia, where control of oak powdery mildew Microsphaera alphitoides Griff. et Maubl. had been conducted through alternative protection measures by means of various dosages of AQ-10 biofungicide, with and without added polymer (which has so far never been used in this country for control of oak powdery mildew). Simultaneous testing was conducted on the efficiency of a chemical sulphur-based preparation (used in this area for many years as a measure of suppression of powdery mildews, without the possibility of developing resistance of the pathogen to the active matter). To date, the Republic of Serbia has registered no fungicides for suppression of pathogens in the forest ecosystems. In order to introduce proper use of new disease-fighting agents into a country, certain relevant principles, requirements and criteria prescribed by the Forest Stewardship Council (FSC) must be observed, primarily with respect to measures of assessment and mitigation of risks, the list of dangerous and highly dangerous pesticides with the possibility of alternative protection. One of the main goals of the research was adjustment of the protective measures to the FSC policy through selection of eco-toxicologically favourable fungicides, given the fact that only preparations named on the list of permitted active matters are approved for use in certified forests. The results of the research have demonstrated that AQ-10 biofungicide can be used as a part of integrated disease management programmes as an alternative, through application of several treatments during vegetation and combination with other active matters registered for these purposes, so as to curtail the use of standard fungicides for control of powdery mildews on oak seedlings in nurseries. The best results in suppression of oak powdery mildew were attained through use of AQ-10 biofungicide (dose 50 or 70g/ha) with added polymer Nu Film-17 (dose 1.0 or 1.5 l/ha). If the treatment is applied at the appropriate time, even fewer number of treatments and smaller doses will be just as efficient.

Keywords: oak powdery mildew, biofungicides, polymers, Microsphaera alphitoides

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2 Microorganisms in Fresh and Stored Bee Pollen Originated from Slovakia

Authors: Vladimíra Kňazovická, Mária Dovičičová, Miroslava Kačániová, Margita Čanigová


The aim of the study was to test the storage of bee pollen at room temperature and in cold store, and to describe microorganisms originated from it. Fresh bee pollen originating in West Slovakia was collected in May 2010. It was tested for presence of particular microbial groups using dilution plating method, and divided into two parts with different storage (in cold store and at room temperature). Microbial analyses of pollen were repeated after one year of storage. Several bacterial strains were isolated and tested using Gram staining, for catalase and fructose-6-phosphate-phosphoketolase presence, and by rapid ID 32A (BioMérieux, France). Micromycetes were identified at genus level. Fresh pollen contained coliform bacteria, which were not detected after one year of storage in both ways. Total plate count (TPC) of aerobes and anaerobes and of yeasts in fresh bee pollen exceeded 5.00 log CFU/g. TPC of aerobes and anaerobes decreased below 2.00 log CFU/g after one year of storage in both ways. Count of yeasts decreased to 2.32 log CFU/g (at room temperature) and to 3.66 log CFU/g (in cold store). Microscopic filamentous fungi decreased from 3.41 log CFU/g (fresh bee pollen) to 1.13 log CFU/g (at room temperature) and to 1.89 log CFU/g (in cold store). In fresh bee pollen, 12 genera of micromycetes were identified in the following order according to their relative density: Penicillium > Mucor > Absidia > Cladosporium, Fusarium > Alternaria > Eurotium > Aspergillus, Rhizopus > Emericella > Arthrinium and Mycelium sterilium. After one year at room temperature, only three genera were detected in bee pollen (Penicillium > Aspergillus, Mucor) and after one year in cold store, seven genera were detected (Mucor > Penicillium, Emericella > Aspergillus, Absidia > Arthrinium, Eurotium). From the plates designated for anaerobes, eight colonies originating in fresh bee pollen were isolated. Among them, a single yeast isolate occurred. Other isolates were G+ bacteria, with a total of five rod shaped. In three out of these five, catalase was absent and fructose-6-phosphate-phosphoketolase was present. Bacterial isolates originating in fresh pollen belonged probably to genus Bifidobacterium or relative genera, but their identity was not confirmed unequivocally. In general, cold conditions are suitable for maintaining the natural properties of foodstuffs for a longer time. Slight decrease of microscopic fungal number and diversity was recorded in cold temperatures compared with storage at room temperature.

Keywords: bacteria, bee product, microscopic fungi, biosystems engineering

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1 Quantified Metabolomics for the Determination of Phenotypes and Biomarkers across Species in Health and Disease

Authors: Miroslava Cuperlovic-Culf, Lipu Wang, Ketty Boyle, Nadine Makley, Ian Burton, Anissa Belkaid, Mohamed Touaibia, Marc E. Surrette


Metabolic changes are one of the major factors in the development of a variety of diseases in various species. Metabolism of agricultural plants is altered the following infection with pathogens sometimes contributing to resistance. At the same time, pathogens use metabolites for infection and progression. In humans, metabolism is a hallmark of cancer development for example. Quantified metabolomics data combined with other omics or clinical data and analyzed using various unsupervised and supervised methods can lead to better diagnosis and prognosis. It can also provide information about resistance as well as contribute knowledge of compounds significant for disease progression or prevention. In this work, different methods for metabolomics quantification and analysis from Nuclear Magnetic Resonance (NMR) measurements that are used for investigation of disease development in wheat and human cells will be presented. One-dimensional 1H NMR spectra are used extensively for metabolic profiling due to their high reliability, wide range of applicability, speed, trivial sample preparation and low cost. This presentation will describe a new method for metabolite quantification from NMR data that combines alignment of spectra of standards to sample spectra followed by multivariate linear regression optimization of spectra of assigned metabolites to samples’ spectra. Several different alignment methods were tested and multivariate linear regression result has been compared with other quantification methods. Quantified metabolomics data can be analyzed in the variety of ways and we will present different clustering methods used for phenotype determination, network analysis providing knowledge about the relationships between metabolites through metabolic network as well as biomarker selection providing novel markers. These analysis methods have been utilized for the investigation of fusarium head blight resistance in wheat cultivars as well as analysis of the effect of estrogen receptor and carbonic anhydrase activation and inhibition on breast cancer cell metabolism. Metabolic changes in spikelet’s of wheat cultivars FL62R1, Stettler, MuchMore and Sumai3 following fusarium graminearum infection were explored. Extensive 1D 1H and 2D NMR measurements provided information for detailed metabolite assignment and quantification leading to possible metabolic markers discriminating resistance level in wheat subtypes. Quantification data is compared to results obtained using other published methods. Fusarium infection induced metabolic changes in different wheat varieties are discussed in the context of metabolic network and resistance. Quantitative metabolomics has been used for the investigation of the effect of targeted enzyme inhibition in cancer. In this work, the effect of 17 β -estradiol and ferulic acid on metabolism of ER+ breast cancer cells has been compared to their effect on ER- control cells. The effect of the inhibitors of carbonic anhydrase on the observed metabolic changes resulting from ER activation has also been determined. Metabolic profiles were studied using 1D and 2D metabolomic NMR experiments, combined with the identification and quantification of metabolites, and the annotation of the results is provided in the context of biochemical pathways.

Keywords: metabolic biomarkers, metabolic network, metabolomics, multivariate linear regression, NMR quantification, quantified metabolomics, spectral alignment

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