Search results for: molecular detection

Authors: Ellana Boada, Eric Santos-Clotas, Alba Cabrera-Codony, Maria Martin, Lluis Baneras, Frederic Gich

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Volatile methylsiloxanes (VMS) are a group of manmade silicone compounds widely used in household and industrial applications that end up on the biogas produced through the anaerobic digestion of organic matter in landfills and wastewater treatment plants. The presence of VMS during the biogas energy conversion can cause damage on the engines, reducing the efficiency of this renewable energy source. Non regenerative adsorption onto activated carbon is the most widely used technology to remove siloxanes from biogas, while new trends point out that biotechnology offers a low-cost and environmentally friendly alternative to conventional technologies. The first objective of this research was to enrich, isolate and identify bacterial species able to grow using siloxane molecules as a sole carbon source: anoxic wastewater sludge was used as initial inoculum in liquid anoxic enrichments, adding D4 (as representative siloxane compound) previously adsorbed on activated carbon. After several months of acclimatization, liquid enrichments were plated onto solid media containing D4 and thirty-four bacterial isolates were obtained. 16S rRNA gene sequencing allowed the identification of strains belonging to the following species: Ciceribacter lividus, Alicycliphilus denitrificans, Pseudomonas aeruginosa and Pseudomonas citronellolis which are described to be capable to degrade toxic volatile organic compounds. Kinetic assays with 8 representative strains revealed higher cell growth in the presence of D4 compared to the control. Our second objective was to characterize the community composition and diversity of the microbial community present in the enrichments and to elucidate whether the isolated strains were representative members of the community or not. DNA samples were extracted, the 16S rRNA gene was amplified (515F & 806R primer pair), and the microbiome analyzed from sequences obtained with a MiSeq PE250 platform. Results showed that the retrieved isolates only represented a minor fraction of the microorganisms present in the enrichment samples, which were represented by Alpha, Beta, and Gamma proteobacteria as dominant groups in the category class thus suggesting that other microbial species and/or consortia may be important for D4 biodegradation. These results highlight the need of additional protocols for the isolation of relevant D4 degraders. Currently, we are developing molecular tools targeting key genes involved in siloxane biodegradation to identify and quantify the capacity of the isolates to metabolize D4 in batch cultures supplied with a synthetic gas stream of air containing 60 mg m⁻³ of D4 together with other volatile organic compounds found in the biogas mixture (i.e. toluene, hexane and limonene). The isolates were used as inoculum in a biotrickling filter containing lava rocks and activated carbon to assess their capacity for siloxane removal. Preliminary results of biotrickling filter performance showed 35% of siloxane biodegradation in a contact time of 14 minutes, denoting that biological siloxane removal is a promising technology for biogas upgrading.

Keywords: bacterial cultivation, biogas upgrading, microbiome, siloxanes

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Authors: Orkide Donma, Mustafa M. Donma, Muhammet Demirkol, Murat Aydin, Tuba Gokkus, Burcin Nalbantoglu, Aysin Nalbantoglu, Birol Topcu

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Obesity in childhood establishes a ground for adulthood obesity. Especially morbid obesity is an important problem for the children because of the associated diseases such as diabetes mellitus, cancer and cardiovascular diseases. In this study, body mass index (BMI), body fat ratios, anthropometric measurements and ratios were evaluated together with different laboratory indices upon evaluation of obesity in morbidly obese (MO) children. Children with nutritional problems participated in the study. Written informed consent was obtained from the parents. Study protocol was approved by the Ethics Committee. Sixty-two MO girls aged 129.5±35.8 months and 75 MO boys aged 120.1±26.6 months were included into the scope of the study. WHO-BMI percentiles for age-and-sex were used to assess the children with those higher than 99^th as morbid obesity. Anthropometric measurements of the children were recorded after their physical examination. Bio-electrical impedance analysis was performed to measure fat distribution. Anthropometric ratios, body fat ratios, Index-I and Index-II as well as insulin sensitivity indices (ISIs) were calculated. Girls as well as boys were binary grouped according to homeostasis model assessment-insulin resistance (HOMA-IR) index of <2.5 and >2.5, fasting glucose to insulin ratio (FGIR) of <6 and >6 and quantitative insulin sensitivity check index (QUICKI) of <0.33 and >0.33 as the frequently used cut-off points. They were evaluated based upon their BMIs, arms, legs, trunk, whole body fat percentages, body fat ratios such as fat mass index (FMI), trunk-to-appendicular fat ratio (TAFR), whole body fat ratio (WBFR), anthropometric measures and ratios [waist-to-hip, head-to-neck, thigh-to-arm, thigh-to-ankle, height/2-to-waist, height/2-to-hip circumference (C)]. SPSS/PASW 18 program was used for statistical analyses. p≤0.05 was accepted as statistically significance level. All of the fat percentages showed differences between below and above the specified cut-off points in girls when evaluated with HOMA-IR and QUICKI. Differences were observed only in arms fat percent for HOMA-IR and legs fat percent for QUICKI in boys (p≤ 0.05). FGIR was unable to detect any differences for the fat percentages of boys. Head-to-neck C was the only anthropometric ratio recommended to be used for all ISIs (p≤0.001 for both girls and boys in HOMA-IR, p≤0.001 for girls and p≤0.05 for boys in FGIR and QUICKI). Indices which are recommended for use in both genders were Index-I, Index-II, HOMA/BMI and log HOMA (p≤0.001). FMI was also a valuable index when evaluated with HOMA-IR and QUICKI (p≤0.001). The important point was the detection of the severe significance for HOMA/BMI and log HOMA while they were evaluated also with the other indices, FGIR and QUICKI (p≤0.001). These parameters along with Index-I were unique at this level of significance for all children. In conclusion, well-accepted ratios or indices may not be valid for the evaluation of both genders. This study has emphasized the limiting properties for boys. This is particularly important for the selection process of some ratios and/or indices during the clinical studies. Gender difference should be taken into consideration for the evaluation of the ratios or indices, which will be recommended to be used particularly within the scope of obesity studies.

Keywords: anthropometry, childhood obesity, gender, insulin sensitivity index

Procedia PDF Downloads 335

Authors: Gerd Wilsch, Tobias Guenther, Tobias Voelker

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In view of the ageing of vital infrastructure facilities, a reliable condition assessment of concrete structures is becoming of increasing interest for asset owners to plan timely and appropriate maintenance and repair interventions. For concrete structures, reinforcement corrosion induced by penetrating chlorides is the dominant deterioration mechanism affecting the serviceability and, eventually, structural performance. The determination of the quantitative chloride ingress is required not only to provide valuable information on the present condition of a structure, but the data obtained can also be used for the prediction of its future development and associated risks. At present, wet chemical analysis of ground concrete samples by a laboratory is the most common test procedure for the determination of the chloride content. As the chloride content is expressed by the mass of the binder, the analysis should involve determination of both the amount of binder and the amount of chloride contained in a concrete sample. This procedure is laborious, time-consuming, and costly. The chloride profile obtained is based on depth intervals of 10 mm. LIBS is an economically viable alternative providing chloride contents at depth intervals of 1 mm or less. It provides two-dimensional maps of quantitative element distributions and can locate spots of higher concentrations like in a crack. The results are correlated directly to the mass of the binder, and it can be applied on-site to deliver instantaneous results for the evaluation of the structure. Examples for the application of the method in the laboratory for the investigation of diffusion and migration of chlorides, sulfates, and alkalis are presented. An example for the visualization of the Li transport in concrete is also shown. These examples show the potential of the method for a fast, reliable, and automated two-dimensional investigation of transport processes. Due to the better spatial resolution, more accurate input parameters for model calculations are determined. By the simultaneous detection of elements such as carbon, chlorine, sodium, and potassium, the mutual influence of the different processes can be determined in only one measurement. Furthermore, the application of a mobile LIBS system in a parking garage is demonstrated. It uses a diode-pumped low energy laser (3 mJ, 1.5 ns, 100 Hz) and a compact NIR spectrometer. A portable scanner allows a two-dimensional quantitative element mapping. Results show the quantitative chloride analysis on wall and floor surfaces. To determine the 2-D distribution of harmful elements (Cl, C), concrete cores were drilled, split, and analyzed directly on-site. Results obtained were compared and verified with laboratory measurements. The results presented show that the LIBS method is a valuable addition to the standard procedures - the wet chemical analysis of ground concrete samples. Currently, work is underway to develop a technical code of practice for the application of the method for the determination of chloride concentration in concrete.

Keywords: chemical analysis, concrete, LIBS, spectroscopy

Procedia PDF Downloads 88

Authors: Anjali Singh, Ashwani Raju, Vandana Devi, Mohmad Mohsin Atique, Satyendra Singh, Munendra Singh

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India is the biggest consumer of mercury and, consequently, a major emitter too. The increasing mercury contamination in India’s water resources has gained widespread attention and, therefore, atmospheric deposition is of critical concern. However, little emphasis was placed on the role of precipitation in the aquatic mercury cycle of the Ganga Alluvial Plain which provides drinking water to nearly 7% of the world’s human population. A majority of the precipitation here occurs primarily in 10% duration of the year in the monsoon season. To evaluate the sources and transportation of mercury, water sample analysis has been conducted from two selected sites near Lucknow, which have a strong hydraulic gradient towards the river. 31 groundwater samples from Jehta village (26°55’15’’N; 80°50’21’’E; 119 m above mean sea level) and 31 river water samples from the Behta Nadi (a tributary of the Gomati River draining into the Ganga River) were collected during the monsoon season on every alternate day between 01 July to 30 August 2019. The total mercury analysis was performed by using Flow Injection Atomic Absorption Spectroscopy (AAS)-Mercury Hybride System, and daily rainfall data was collected from the India Meteorological Department, Amausi, Lucknow. The ambient groundwater and river-water concentrations were both 2-4 ng/L as there is no known geogenic source of mercury found in the area. Before the onset of the monsoon season, the groundwater and the river-water recorded mercury concentrations two orders of magnitude higher than the ambient concentrations, indicating the regional transportation of the mercury from the non-point source into the aquatic environment. Maximum mercury concentrations in groundwater and river-water were three orders of magnitude higher than the ambient concentrations after the onset of the monsoon season characterizing the considerable mobilization and redistribution of mercury by monsoonal precipitation. About 50% of both of the water samples were reported mercury below the detection limit, which can be mostly linked to the low intensity of precipitation in August and also with the dilution factor by precipitation. The highest concentration ( > 1200 ng/L) of mercury in groundwater was reported after 6-days lag from the first precipitation peak. Two high concentration peaks (>1000 ng/L) in river-water were separately correlated with the surface flow and groundwater outflow of mercury. We attribute the elevated mercury concentration in both of the water samples before the precipitation event to mercury originating from the extensive use of agrochemicals in mango farming in the plain. However, the elevated mercury concentration during the onset of monsoon appears to increase in area wetted with atmospherically deposited mercury, which migrated down from surface water to groundwater as downslope migration is a fundamental mechanism seen in rivers of the alluvial plain. The present study underscores the significance of monsoonal precipitation in the transportation of mercury to drinking water resources of the Ganga Alluvial Plain. This study also suggests that future research must be pursued for a better understand of the human health impact of mercury contamination and for quantification of the role of Ganga Alluvial Plain in the Global Mercury Cycle.

Keywords: drinking water resources, Ganga alluvial plain, india, mercury

Procedia PDF Downloads 124

Authors: Andrés Quiroz, Emilio Hormazábal, Ana Mutis, Fernando Ortega, Loreto Méndez, Leonardo Parra

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Red clover root borer, Hylastinus obscurus Marsham (Coleoptera: Curculionidae), is the main insect pest associated to red clover, Trifolium pratense L. An average of 1.5 H. obscurus per plant can cause 5.5% reduction in forage yield in pastures of two to three years old. Moreover, insect attack can reach 70% to 100% of the plants. To our knowledge, there is no a chemical strategy for controlling this pest. Therefore alternative strategies for controlling H. obscurus are a high priority for red clover producers. One of this alternative is related to the study of secondary metabolites involved in intrinsic chemical defenses developed by plants, such as isoflavonoids. The isoflavonoids formononetin and daidzein have elicited an antifeedant and phagostimult effect on H. obscurus respectively. However, we do not know how is the dynamic variation of these isoflavonoids under field conditions. The main objective of this work was to evaluate the variation of the antifeedant isoflavonoids formononetin, the phagostimulant isoflavonoids daidzein, and their respective glycosides over time in different ecotypes of red clover. Fourteen red clover ecotypes (8 cultivars and 6 experimental lines), were collected at INIA-Carillanca (La Araucanía, Chile). These plants were established in October 2015 under irrigated conditions. The cultivars were distributed in a randomized complete block with three replicates. The whole plants were sampled in four times: 15th October 2016, 12th December 2016, 27th January 2017 and 16th March 2017 with sufficient amount of soil to avoid root damage. A polar fraction of isoflavonoid was obtained from 20 mg of lyophilized root tissue extracted with 2 mL of 80% MeOH for 16 h using an orbital shaker in the dark at room temperature. After, an aliquot of 1.4 mL of the supernatant was evaporated, and the residue was resuspended in 300 µL of 45% MeOH. The identification and quantification of isoflavonoid root extracts were performed by the injection of 20 µL into a Shimadzu HPLC equipped with a C-18 column. The sample was eluted with a mobile phase composed of AcOH: H₂O (1:9 v/v) as solvent A and CH₃CN as solvent B. The detection was performed at 260 nm. The results showed that the amount of aglycones was higher than the respective glycosides. This result is according to the biosynthetic pathway of flavonoids, where the formation of glycoside is further to the glycosides biosynthesis. The amount of formononetin was higher than daidzein. In roots, where H. obscurus spent the most part of its live cycle, the highest content of formononetin was found in G 27, Pawera, Sabtoron High, Redqueli-INIA and Superqueli-INIA cvs. (2.1, 1.8, 1.8, 1.6 and 1.0 mg g⁻¹ respectively); and the lowest amount of daidzein were found Superqueli-INIA (0.32 mg g⁻¹) and in the experimental line Sel Syn Int4 (0.24 mg g⁻¹). This ecotype showed a high content of formononetin (0.9 mg g⁻¹). This information, associated with cultural practices, could help farmers and breeders to reduce H. obscurus in grassland, selecting ecotypes with high content of formononetin and low amount of daidzein in the roots of red clover plants. Acknowledgements: FONDECYT 1141245 and 11130715.

Keywords: daidzein, formononetin, isoflavonoid glycosides, trifolium pratense

Procedia PDF Downloads 191

Authors: R. R. Galiev, I. I. Lykov, A. E. Shitikov, I. A. Bilenko

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Dual-comb interferometry is based on the mixing of two optical frequency combs with slightly different lines spacing which results in the mapping of the optical spectrum into the radio-frequency domain for future digitizing and numerical processing. The dual-comb approach enables diverse applications, including metrology, fast high-precision spectroscopy, and distance range. Ordinary frequency-modulated continuous-wave (FMCW) laser-based Light Identification Detection and Ranging systems (LIDARs) suffer from two main disadvantages: slow and unreliable mechanical, spatial scan and a rather wide linewidth of conventional lasers, which limits speed measurement resolution. Dual-comb distance measurements with Allan deviations down to 12 nanometers at averaging times of 13 microseconds, along with ultrafast ranging at acquisition rates of 100 megahertz, allowing for an in-flight sampling of gun projectiles moving at 150 meters per second, was previously demonstrated. Nevertheless, pump lasers with EDFA amplifiers made the device bulky and expensive. An alternative approach is a direct coupling of the laser to a reference microring cavity. Backscattering can tune the laser to the eigenfrequency of the cavity via the so-called self-injection locked (SIL) effect. Moreover, the nonlinearity of the cavity allows a solitonic frequency comb generation in the very same cavity. In this work, we developed a fully integrated, power-efficient, electrically driven dual-micro comb source based on the semiconductor lasers SIL to high-quality integrated Si3N4 microresonators. We managed to obtain robust 1400-1700 nm combs generation with a 150 GHz or 1 THz lines spacing and measure less than a 1 kHz Lorentzian withs of stable, MHz spaced beat notes in a GHz band using two separated chips, each pumped by its own, self-injection locked laser. A deep investigation of the SIL dynamic allows us to find out the turn-key operation regime even for affordable Fabry-Perot multifrequency lasers used as a pump. It is important that such lasers are usually more powerful than DFB ones, which were also tested in our experiments. In order to test the advantages of the proposed techniques, we experimentally measured a minimum detectable speed of a reflective object. It has been shown that the narrow line of the laser locked to the microresonator provides markedly better velocity accuracy, showing velocity resolution down to 16 nm/s, while the no-SIL diode laser only allowed 160 nm/s with good accuracy. The results obtained are in agreement with the estimations and open up ways to develop LIDARs based on compact and cheap lasers. Our implementation uses affordable components, including semiconductor laser diodes and commercially available silicon nitride photonic circuits with microresonators.

Keywords: dual-comb spectroscopy, LIDAR, optical microresonator, self-injection locking

Procedia PDF Downloads 49

Authors: Hannah R. Devens, Phillip L. Davidson, Dione Deaker, Kathryn E. Smith, Gregory A. Wray, Maria Byrne

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Marine invertebrate species with calcifying larvae are especially vulnerable to ocean acidification (OA) caused by rising atmospheric CO₂ levels. Acidic conditions can delay development, suppress metabolism, and decrease the availability of carbonate ions in the ocean environment for skeletogenesis. These stresses often result in increased larval mortality, which may lead to significant ecological consequences including alterations to the larval settlement, population distribution, and genetic connectivity. Importantly, many of these physiological and developmental effects are caused by genetic and molecular level changes. Although many studies have examined the effect of near-future oceanic pH levels on gene expression in marine invertebrates, little is known about the impact of OA on gene expression in a developmental context. Here, we performed mRNA-sequencing to investigate the impact of environmental acidity on gene expression across three developmental stages in the sea urchin Heliocidaris erythrogramma. We collected RNA from gastrula, early larva, and 1-day post-metamorphic juvenile sea urchins cultured at present-day and predicted future oceanic pH levels (pH 8.1 and 7.7, respectively). We assembled an annotated reference transcriptome encompassing development from egg to ten days post-metamorphosis by combining these data with datasets from two previous developmental transcriptomic studies of H. erythrogramma. Differential gene expression and time course analyses between pH conditions revealed significant alterations to developmental transcription that are potentially associated with pH stress. Consistent with previous investigations, genes involved in biomineralization and ion transport were significantly upregulated under acidic conditions. Differences in gene expression between the two pH conditions became more pronounced post-metamorphosis, suggesting a development-dependent effect of OA on gene expression. Furthermore, many differences in gene expression later in development appeared to be a result of broad downregulation at pH 7.7: of 539 genes differentially expressed at the juvenile stage, 519 of these were lower in the acidic condition. Time course comparisons between pH 8.1 and 7.7 samples also demonstrated over 500 genes were more lowly expressed in pH 7.7 samples throughout development. Of the genes exhibiting stage-dependent expression level changes, over 15% of these diverged from the expected temporal pattern of expression in the acidic condition. Through these analyses, we identify novel candidate genes involved in development, metabolism, and transcriptional regulation that are possibly affected by pH stress. Our results demonstrate that pH stress significantly alters gene expression dynamics throughout development. A large number of genes differentially expressed between pH conditions in juveniles relative to earlier stages may be attributed to the effects of acidity on transcriptional regulation, as a greater proportion of mRNA at this later stage has been nascent transcribed rather than maternally loaded. Also, the overall downregulation of many genes in the acidic condition suggests that OA-induced developmental delay manifests as suppressed mRNA expression, possibly from lower transcription rates or increased mRNA degradation in the acidic environment. Further studies will be necessary to determine in greater detail the extent of OA effects on early developing marine invertebrates.

Keywords: development, gene expression, ocean acidification, RNA-sequencing, sea urchins

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Authors: Vandana Mohan, Ashwani Koul

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Aim: To evaluate the potential of Aqueous Tinospora cordifolia stem extract (Aq.Tc) and Arabinogalactan (AG) on pulmonary carcinogenesis and associated tumor markers. Background: Lung cancer is one of the most frequent malignancy with high mortality rate due to limitation of early detection resulting in low cure rates. Current research effort focuses on identifying some blood-based biomarkers like CEA, ctDNA and LDH which may have potential to detect cancer at an early stage, evaluation of therapeutic response and its recurrence. Medicinal plants and their active components have been widely investigated for their anticancer potentials. Aqueous preparation of T. Cordifolia extract is enriched in the polysaccharide fraction i.e., AG when compared with other types of extract. Moreover, reports are available of polysaccharide fraction of T. Cordifolia in in vitro lung cancer models which showed profound anti-metastatic activity against these cell lines. However, not much has been explored about its effect in in vivo lung cancer models and the underlying mechanism involved. Experimental Design: Mice were randomly segregated into six groups. Group I animals served as control. Group II animals were administered with Aq. Tc extract (200 mg/kg b.w.) p.o.on the alternate days. Group III animals were fed with AG (7.5 mg/kg b.w.) p.o. on the alternate days (thrice a week). Group IV animals were installed with Benzo(a)pyrene (50 mg/kg b.w.), i.p. twice within an interval of two weeks. Group V animals received Aq. Tc extract as in group II along with it B(a)P was installed after two weeks of Aq. Tc administration following the same protocol as for group IV. Group VI animals received AG as in group III along with it B(a)P was installed after two weeks of AG administration. Results: Administration of B(a)P to mice resulted in increased tumor incidence, multiplicity and pulmonary somatic index with concomitant increase in serum/plasma markers like CEA, ctDNA, LDH and TNF-α.Aq.Tc and AG supplementation significantly attenuated these alterations at different stages of tumorigenesis thereby showing potent anti-cancer effect in lung cancer. A pronounced decrease in serum/plasma markers were observed in animals treated with Aq.Tc as compared to those fed with AG. Also, extensive hyperproliferation of alveolar epithelium was prominent in B(a)P induced lung tumors. However, treatment of Aq.Tc and AG to lung tumor bearing mice exhibited reduced alveolar damage evident from decreased number of hyperchromatic irregular nuclei. A direct correlation between the concentration of tumor markers and the intensity of lung cancer was observed in animals bearing cancer co-treated with Aq.Tc and AG. Conclusion: These findings substantiate the chemopreventive potential of Aq.Tc and AG against lung tumorigenesis. Interestingly, Aq.Tc was found to be more effective in modulating the cancer as reflected by various observations which may be attributed to the synergism offered by various components of Aq.Tc. Further studies are in progress to understand the underlined mechanism in inhibiting lung tumorigenesis by Aq.Tc and AG.

Keywords: Arabinogalactan, Benzo(a)pyrene B(a)P, carcinoembryonic antigen (CEA), circulating tumor DNA (ctDNA), lactate dehydrogenase (LDH), Tinospora cordifolia

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Authors: Olga Smilevska Spasova, Katerina Boshkovska, Gorica Popova, Mirjana Popovska

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Introduction: Pneumonia is an infection of the pulmonary parenchyma. HPIV 3 is one of four viruses that is a member of the Paramyxoviridae family designated types 1-4 that have a nonsegmented, single-stranded RNA genome with a lipid-containing envelope. They are spread from the respiratory tract by aerosolized secretions or by direct contact with secretions. Type 3 is endemic and can cause serious illness in immunocompromised patients. Illness caused by parainfluenza occurs shortly after inoculation with the virus. The level of immunoglobulin A antibody in serum is the best predictor of susceptibility to infection. Streptococcus pneumonia or pneumococcus is a Gram-positive, spherical bacteria, usually found in pairs and it is a member of the genus Streptococcus. Streptococcus pneumonia resides asymptomatically in healthy carriers typically colonizing the respiratory tract, sinuses, and nasal cavity. In individuals with weaker immune systems like young infants, pneumococcal bacterium is the most common cause of community-acquired pneumonia in the world. Case Report: The aim is to present a case of lower respiratory tract infection in an infant caused by parainfluenza virus 3, S. pneumonia and undifferentiated gram-negative bacteria that was successfully treated. The infant is with a history of recurrent episodes of wheezing in the past 3mounts.Infant of 10months presents 2weeks before admittance with high fever, runny nose, and cough. The primary pediatrician prescribed oral cefpodoxime for 10days and inhaled salbutamol. Two days before admittance in hospital the infant with high fever, cough, and difficulty breathing. At admittance, infant is pale, anxious with rapid respirations, cough, wheezing and tachycardia. On auscultation: vesicular breathing sounds with high pitched wheezing and on the right coarse crackles. Investigations: Blood analysis: RBC: 4, 7 x1012L, WBC: 8,3x109L: Neut: 42.73% Lym: 41.57%, Hgb: 9.38 g/dl MCV: 62.7fl, MCH: 20.0pg MCHC: 31.8 g/dl RDW: 18.7% Plt-307.9 x109LCRP: 2,5mg/l, serum iron-7.92umol/l, O2sat-97% on blood gas analysis, puls-125/min.X-ray of chest with hyperinflationand right pericardial consolidation. Microbiological analysis of sputum sample is positive for undifferentiated gram-negative bacteria (colonizer)–resistant to cefotaxime, ampicillin, cefoxitin, sulfamet.+trimetoprim and sensitive to amikacin, gentamicin, and ciprofloxacin. Molecular multiplex RT-PCR for 19 viruses and multiplex PCR for 7 bacteria test for respiratory pathogens positive for Parainfluenza virus 3(Ct=22.73), Streptococcus pneumonia (Ct=26.75).IED: IgG-9.31g/l, IgA-0.351g/l, IgM-0.86g/l. Therapy: Treatment was started with inhaled salbutamol, intravenous antibiotic cefotaxime as well as systemic corticosteroids. On day 7 because of slow clinical resolution of chest auscultation findings and an etiologic clue with a positive sputum sample for resistant undifferentiated gram negative bacteria, a second intravenous antibiotic was administered amikacin. The infant is discharged on day 14 with resolution of clinical findings. Conclusion: Mixed co-infections with respiratory viruses and bacteria in immunocompromised infants are likely to lead to a more severe form of community acquired pneumonia that will need hospitalization.

Keywords: HPIV- 3, infant, pneumonia, S. pneumonia, x-ray chest

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Authors: A. Cittadino, P. Gantes, C. Coviella, M. Casset, A. Sanchez Caro

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Three currently active landfills receive the waste from Buenos Aires city and the Great Buenos Aires suburbs. One of the first landfills to receive solid waste from this area was located in Villa Dominico, some 7 km south from Buenos Aires City. With an area of some 750 ha, including riparian habitats, divided into 14 cells, it received solid wastes from June 1979 through February 2004. In December 2010, a biological monitoring program was set up by CEAMSE and Universidad Nacional de Lujan, still operational to date. The aim of the monitoring program is to assess the state of several biological groups within the landfill and to follow their dynamics overtime in order to identify if any, early signs of damage the landfill activities might have over the biota present. Bird and rodent populations, aquatic and terrestrial invertebrates’ populations, cells vegetation coverage, and surrounding areas vegetation coverage and main composition are followed by quarterly samplings. Bird species richness and abundance were estimated by observation over walk transects on each environment. A total of 74 different species of birds were identified. Species richness and diversity were high for both riparian surrounding areas and within the landfill. Several grassland -typical of the 'Pampa'- bird species were found within the landfill, as well as some migratory and endangered bird species. Sherman and Tomahawk traps are set overnight for small mammal sampling. Rodent populations are just above detection limits, and the few specimens captured belong mainly to species common to rural areas, instead of city-dwelling species. The two marsupial species present in the region were captured on occasions. Aquatic macroinvertebrates were sampled on a watercourse upstream and downstream the outlet of the landfill’s wastewater treatment plant and are used to follow water quality using biological indices. Water quality ranged between weak and severe pollution; benthic invertebrates sampled before and after the landfill, show no significant differences in water quality using the IBMWP index. Insect biota from yellow sticky cards and pitfall traps showed over 90 different morphospecies, with Shannon diversity index running from 1.9 to 3.9, strongly affected by the season. An easy-to-perform non-expert demandant method was used to assess vegetation coverage. Two scales of determination are utilized: field observation (1 m resolution), and Google Earth images (that allow for a better than 5 m resolution). Over the eight year period of the study, vegetation coverage over the landfill cells run from a low 83% to 100% on different cells, with an average between 95 to 99% for the entire landfill depending on seasonality. Surrounding area vegetation showed almost 100% coverage during the entire period, with an average density from 2 to 6 species per sq meter and no signs of leachate damaged vegetation.

Keywords: biological indicators, biota monitoring, landfill species diversity, waste management

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Authors: Artem Kim, Clara Savary, Christele Dubourg, Wilfrid Carre, Houda Hamdi-Roze, Valerie Dupé, Sylvie Odent, Marie De Tayrac, Veronique David

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Holoprosencephaly (HPE) is a rare congenital brain malformation resulting from the incomplete separation of the two cerebral hemispheres. It is characterized by a wide phenotypic spectrum and a high degree of locus heterogeneity. Genetic defects in 16 genes have already been implicated in HPE, but account for only 30% of cases, suggesting that a large part of genetic factors remains to be discovered. HPE has been recently redefined as a complex multigenic disorder, requiring the joint effect of multiple mutational events in genes belonging to one or several developmental pathways. The onset of HPE may result from accumulation of the effects of multiple rare variants in functionally-related genes, each conferring a moderate increase in the risk of HPE onset. In order to decipher the genetic basis of HPE, unconventional patterns of inheritance involving multiple genetic factors need to be considered. The primary objective of this study was to uncover possible disease causing combinations of multiple rare variants underlying HPE by performing trio-based Whole Exome Sequencing (WES) of familial cases where no molecular diagnosis could be established. 39 families were selected with no fully-penetrant causal mutation in known HPE gene, no chromosomic aberrations/copy number variants and without any implication of environmental factors. As the main challenge was to identify disease-related variants among a large number of nonpathogenic polymorphisms detected by WES classical scheme, a novel variant prioritization approach was established. It combined WES filtering with complementary gene-level approaches: transcriptome-driven (RNA-Seq data) and clinically-driven (public clinical data) strategies. Briefly, a filtering approach was performed to select variants compatible with disease segregation, population frequency and pathogenicity prediction to identify an exhaustive list of rare deleterious variants. The exome search space was then reduced by restricting the analysis to candidate genes identified by either transcriptome-driven strategy (genes sharing highly similar expression patterns with known HPE genes during cerebral development) or clinically-driven strategy (genes associated to phenotypes of interest overlapping with HPE). Deeper analyses of candidate variants were then performed on a family-by-family basis. These included the exploration of clinical information, expression studies, variant characteristics, recurrence of mutated genes and available biological knowledge. A novel bioinformatics pipeline was designed. Applied to the 39 families, this final integrated workflow identified an average of 11 candidate variants per family. Most of candidate variants were inherited from asymptomatic parents suggesting a multigenic inheritance pattern requiring the association of multiple mutational events. The manual analysis highlighted 5 new strong HPE candidate genes showing recurrences in distinct families. Functional validations of these genes are foreseen.

Keywords: complex genetic disorder, holoprosencephaly, multiple rare variants, whole exome sequencing

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Authors: Elvin P. Chizenga, Heidi Abrahamse

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Cancer cell resistance to therapy is the main cause of treatment failures and the poor prognosis of cancer convalescence. The progression of cervical cancer to other parts of the genitourinary system and the reported recurrence rates are overwhelming. Current treatments, including surgery, chemo and radiation have been inefficient in eradicating the tumor cells. These treatments are also associated with poor prognosis and reduced quality of life, including fertility loss. This has inspired the need for the development of new treatment modalities to eradicate cervical cancer successfully. Photodynamic Therapy (PDT) is a modern treatment modality that induces cell death by photochemical interactions of light and a photosensitizer, which in the presence of molecular oxygen, yields a set of chemical reactions that generate Reactive Oxygen Species (ROS) and other free radical species causing cell damage. Enhancing PDT using modified drug delivery can increase the concentration of the photosensitizer in the tumor cells, and this has the potential to maximize its therapeutic efficacy. In cervical cancer, all infected cells constitutively express genes of the E6 and E7 HPV viral oncoproteins, resulting in high concentrations of E6 and E7 in the cytoplasm. This provides an opportunity for active targeting of cervical cancer cells using immune-mediated drug delivery to maximize therapeutic efficacy. The use of nanoparticles in PDT has also proven effective in enhancing therapeutic efficacy. Gold nanoparticles (AuNps) in particular, are explored for their use in biomedicine due to their biocompatibility, low toxicity, and enhancement of drug uptake by tumor cells. In this present study, a biomolecule comprising of AuNPs, anti-E6 monoclonal antibodies, and Aluminium Phthalocyanine photosensitizer was synthesized for use in targeted PDT of cervical cancer. The AuNp-Anti-E6-Sulfonated Aluminium Phthalocyanine mix (AlPcSmix) photosensitizing biomolecule was synthesized by coupling AuNps and anti-E6 monoclonal antibodies to the AlPcSmix via Polyethylene Glycol (PEG) chemical links. The final product was characterized using Transmission Electron Microscope (TEM), Zeta Potential, Uv-Vis Spectrophotometry, Fourier Transform Infrared Spectroscopy (FTIR), and X-ray diffraction (XRD), to confirm its chemical structure and functionality. To observe its therapeutic role in treating cervical cancer, cervical cancer cells, HeLa cells were seeded in 3.4 cm² diameter culture dishes at a concentration of 5x10⁵ cells/ml, in vitro. The cells were treated with varying concentrations of the photosensitizing biomolecule and irradiated using a 673.2 nm wavelength of laser light. Post irradiation cellular responses were performed to observe changes in morphology, viability, proliferation, cytotoxicity, and cell death pathways induced. Dose-Dependent response of the cells to treatment was demonstrated as significant morphologic changes, increased cytotoxicity, and decreased cell viability and proliferation This study presented a synthetic biomolecule for targeted PDT of cervical cancer. The study suggested that PDT using this AuNp- Anti-E6- AlPcSmix photosensitizing biomolecule is a very effective treatment method for the eradication of cervical cancer cells, in vitro. Further studies in vivo need to be conducted to support the use of this biomolecule in treating cervical cancer in clinical settings.

Keywords: anti-E6 monoclonal antibody, cervical cancer, gold nanoparticles, photodynamic therapy

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Authors: Sam Bartlett, Thomas Bacquart, Arul Murugan, Abigail Morris

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Fuel cell electric vehicles provide the potential to decarbonise road transport, create new economic opportunities, diversify national energy supply, and significantly reduce the environmental impacts of road transport. A potential issue, however, is that the catalyst used at the fuel cell cathode is susceptible to degradation by impurities, especially sulphur-containing compounds. A recent European Directive (2014/94/EU) stipulates that, from November 2017, all hydrogen provided to fuel cell vehicles in Europe must comply with the hydrogen purity specifications listed in ISO 14687-2; this includes reactive and toxic chemicals such as ammonia and total sulphur-containing compounds. This requirement poses great analytical challenges due to the instability of some of these compounds in calibration gas standards at relatively low amount fractions and the difficulty associated with undertaking measurements of groups of compounds rather than individual compounds. Without the available reference materials and analytical infrastructure, hydrogen refuelling stations will not be able to demonstrate compliance to the ISO 14687 specifications. The hydrogen purity laboratory at NPL provides world leading, accredited purity measurements to allow hydrogen refuelling stations to evidence compliance to ISO 14687. Utilising state-of-the-art methods that have been developed by NPL’s hydrogen purity laboratory, including a novel method for measuring total sulphur compounds at 4 nmol/mol and a hydrogen impurity enrichment device, we provide the capabilities necessary to achieve these goals. An overview of these capabilities will be given in this paper. As part of the EMPIR Hydrogen co-normative project ‘Metrology for sustainable hydrogen energy applications’, NPL are developing a validated analytical methodology for the measurement of speciated sulphur-containing compounds in hydrogen at low amount fractions pmol/mol to nmol/mol) to allow identification and measurement of individual sulphur-containing impurities in real samples of hydrogen (opposed to a ‘total sulphur’ measurement). This is achieved by producing a suite of stable gravimetrically-prepared primary reference gas standards containing low amount fractions of sulphur-containing compounds (hydrogen sulphide, carbonyl sulphide, carbon disulphide, 2-methyl-2-propanethiol and tetrahydrothiophene have been selected for use in this study) to be used in conjunction with novel dynamic dilution facilities to enable generation of pmol/mol to nmol/mol level gas mixtures (a dynamic method is required as compounds at these levels would be unstable in gas cylinder mixtures). Method development and optimisation are performed using gas chromatographic techniques assisted by cryo-trapping technologies and coupled with sulphur chemiluminescence detection to allow improved qualitative and quantitative analyses of sulphur-containing impurities in hydrogen. The paper will review the state-of-the art gas standard preparation techniques, including the use and testing of dynamic dilution technologies for reactive chemical components in hydrogen. Method development will also be presented highlighting the advances in the measurement of speciated sulphur compounds in hydrogen at low amount fractions.

Keywords: gas chromatography, hydrogen purity, ISO 14687, sulphur chemiluminescence detector

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Authors: Sebastian Hiergeist

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It is a common view that Unmanned Aerial Vehicles (UAV) tend to migrate into the civil airspace. This trend is challenging UAV manufacturer in plenty ways, as there come up a lot of new requirements and functional aspects. On the higher application levels, this might be collision detection and avoidance and similar features, whereas all these functions only act as input for the flight control components of the aircraft. The flight control computer (FCC) is the central component when it comes up to ensure a continuous safe flight and landing. As these systems are flight critical, they have to be built up redundantly to be able to provide a Fail-Operational behavior. Recent architectural approaches of FCCs used in UAV systems are often based on very simple microprocessors in combination with proprietary Application-Specific Integrated Circuit (ASIC) or Field Programmable Gate Array (FPGA) extensions implementing the whole redundancy functionality. In the future, such simple microprocessors may not be available anymore as they are more and more replaced by higher sophisticated System on Chip (SoC). As the avionic industry cannot provide enough market power to significantly influence the development of new semiconductor products, the use of solutions from foreign markets is almost inevitable. Products stemming from the industrial market developed according to IEC 61508, or automotive SoCs, according to ISO 26262, can be seen as candidates as they have been developed for similar environments. Current available SoC from the industrial or automotive sector provides quite a broad selection of interfaces like, i.e., Ethernet, SPI or FlexRay, that might come into account for the implementation of a redundancy network. In this context, possible network architectures shall be investigated which could be established by using the interfaces stated above. Of importance here is the avoidance of any single point of failures, as well as a proper segregation in distinct fault containment regions. The performed analysis is supported by the use of guidelines, published by the aviation authorities (FAA and EASA), on the reliability of data networks. The main focus clearly lies on the reachable level of safety, but also other aspects like performance and determinism play an important role and are considered in the research. Due to the further increase in design complexity of recent and future SoCs, also the risk of design errors, which might lead to common mode faults, increases. Thus in the context of this work also the aspect of dissimilarity will be considered to limit the effect of design errors. To achieve this, the work is limited to broadly available interfaces available in products from the most common silicon manufacturer. The resulting work shall support the design of future UAV FCCs by giving a guideline on building up a redundancy network between SoCs, solely using on board interfaces. Therefore the author will provide a detailed usability analysis on available interfaces provided by recent SoC solutions, suggestions on possible redundancy architectures based on these interfaces and an assessment of the most relevant characteristics of the suggested network architectures, like e.g. safety or performance.

Keywords: redundancy, System-on-Chip, UAV, flight control computer (FCC)

Procedia PDF Downloads 190

Authors: Anita Conti, Riccardo Ossanna, Lindsey A. Quintero, Giamaica Conti, Andrea Sbarbati

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Ischemia/reperfusion (IR) injury induces muscle fiber atrophy and skeletal muscle fiber death with subsequently functionality loss. The heterogeneous pool of cells, especially mesenchymal stem cells, contained in the stromal vascular fraction (SVF) of adipose tissue could promote muscle fiber regeneration. To prevent SVF dispersion, it has been proposed the use of injectable biopolymers that work as cells carrier. A significant element of the extracellular matrix is hyaluronic acid (HA), which has been widely used in regenerative medicine as a cell scaffold given its biocompatibility, degradability, and the possibility of chemical functionalization. Connective tissue micro-fragments enriched with SVF obtained from mechanical disaggregation of adipose tissue were evaluated for IR muscle injury regeneration using low molecular weight HA as a scaffold. IR induction. Hindlimb ischemia was induced in 9 athymic nude mice through the clamping of the right quadriceps using a plastic band. Reperfusion was induced by cutting the plastic band after 3 hours of ischemic period. Contralateral (left) muscular tissue was used as healthy control. Treatment. Twenty-four hours after the IR induction, animals (n=3) were intramuscularly injected with 100 µl of SVF mixed with HA (SVF-HA). Animals treated with 100 µl of HA (n=3) and 100 µl saline solution (n=3) were used as control. Treatment monitoring. All animals were in vivo monitored by magnetic resonance imaging (MRI) at 5, 7, 14 and 18 days post-injury (dpi). High-resolution morphological T2 weighed, quantitative T2 map and Dynamic Contrast-Enhanced (DCE) images were acquired in order to assess the regenerative potential of SVF-HA treatment. Ex vivo evaluation. After 18 days from IR induction, animals were sacrificed, and the muscles were harvested for histological examination. At 5 dpi T2 high-resolution MR images clearly reveal the presence of an extensive edematous area due to IR damage for all groups identifiable as an increase of signal intensity (SI) of muscular and surrounding tissue. At 7 dpi, animals of the SVF-HA group showed a reduction of SI, and the T2relaxation time of muscle tissue of the HA-SVF group was 29±0.5ms, comparable with the T2relaxation time of contralateral muscular tissue (30±0.7ms). These suggest a reduction of edematous overflow and swelling. The T2relaxation time at 7dpi of HA and saline groups were 84±2ms and 90±5ms, respectively, which remained elevated during the rest of the study. The evaluation of vascular regeneration showed similar results. Indeed, DCE-MRI analysis revealed a complete recovery of muscular tissue perfusion after 14 dpi for the SVF-HA group, while for the saline and HA group, controls remained in a damaged state. Finally, the histological examination of SVF-HA treated animals exhibited well-defined and organized fibers morphology with a lateralized nucleus, similar to contralateral healthy muscular tissue. On the contrary, HA and saline-treated animals presented inflammatory infiltrates, with HA slightly improving the diameter of the fibers and less degenerated tissue. Our findings show that connective tissue micro-fragments enriched with SVF induce higher muscle homeostasis and perfusion restoration in contrast to control groups.

Keywords: ischemia/reperfusion injury, regenerative medicine, resonance imaging, stromal vascular fraction

Procedia PDF Downloads 92

Authors: Michela Del Vicario, Walter Quattrociocchi, Antonio Scala, Ana Lucía Schmidt, Fabiana Zollo

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Information, rumors, and debates may shape and impact public opinion heavily. In the latest years, several concerns have been expressed about social influence on the Internet and the outcome that online debates might have on real-world processes. Indeed, on online social networks users tend to select information that is coherent to their system of beliefs and to form groups of like-minded people –i.e., echo chambers– where they reinforce and polarize their opinions. In this way, the potential benefits coming from the exposure to different points of view may be reduced dramatically, and individuals' views may become more and more extreme. Such a context fosters misinformation spreading, which has always represented a socio-political and economic risk. The persistence of unsubstantiated rumors –e.g., the hypothetical and hazardous link between vaccines and autism– suggests that social media do have the power to misinform, manipulate, or control public opinion. As an example, current approaches such as debunking efforts or algorithmic-driven solutions based on the reputation of the source seem to prove ineffective against collective superstition. Indeed, experimental evidence shows that confirmatory information gets accepted even when containing deliberately false claims while dissenting information is mainly ignored, influences users’ emotions negatively and may even increase group polarization. Moreover, confirmation bias has been shown to play a pivotal role in information cascades, posing serious warnings about the efficacy of current debunking efforts. Nevertheless, mitigation strategies have to be adopted. To generalize the problem and to better understand social dynamics behind information spreading, in this work we rely on a tight quantitative analysis to investigate the behavior of more than 300M users w.r.t. news consumption on Facebook over a time span of six years (2010-2015). Through a massive analysis on 920 news outlets pages, we are able to characterize the anatomy of news consumption on a global and international scale. We show that users tend to focus on a limited set of pages (selective exposure) eliciting a sharp and polarized community structure among news outlets. Moreover, we find similar patterns around the Brexit –the British referendum to leave the European Union– debate, where we observe the spontaneous emergence of two well segregated and polarized groups of users around news outlets. Our findings provide interesting insights into the determinants of polarization and the evolution of core narratives on online debating. Our main aim is to understand and map the information space on online social media by identifying non-trivial proxies for the early detection of massive informational cascades. Furthermore, by combining users traces, we are finally able to draft the main concepts and beliefs of the core narrative of an echo chamber and its related perceptions.

Keywords: information spreading, misinformation, narratives, online social networks, polarization

Procedia PDF Downloads 267

Authors: Zeyu Zhou, Ziyu Zhao, Xiaochen Yang, Ling AI, Heng Zhai, Stuart Holmes

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As a promising sustainable alternative to traditional fossil fuels, fuel cell technology is highly favoured due to its enhanced working efficiency and reduced emissions. In the context of high-temperature fuel cells (operating above 100 °C), the most commonly used proton exchange membrane (PEM) is the Polybenzimidazole (PBI) doped phosphoric acid (PA) membrane. Grafting is a promising strategy to advance PA-doped PBI PEM technology. The existing grafting modification on PBI PEMs mainly focuses on grafting phosphate-containing or alkaline groups onto the PBI molecular chains. However, quaternary ammonium-based grafting approaches face a common challenge. To initiate the N-alkylation reaction, deacidifying agents such as NaH, NaOH, KOH, K2CO3, etc., can lead to ionic crosslinking between the quaternary ammonium group and PBI. Polyvinylpyrrolidone (PVP) is another widely used polymer, the N-heterocycle groups within PVP endow it with a significant ability to absorb PA. Recently, PVP has attracted substantial attention in the field of fuel cells due to its reduced environmental impact and impressive fuel cell performance. However, due to the the poor compatibility of PVP in PBI, few research apply PVP in PA-doped PBI PEMs. This work introduces an innovative strategy to graft PVP onto PBI to form a network-like polymer. Due to the absence of quaternary ammonium groups, PVP does not pose issues related to crosslinking with PBI. Moreover, the nitrogen-containing functional groups on PVP provide PBI with a robust phosphoric acid retention ability. The nuclear magnetic resonance (NMR) hydrogen spectrum analysis results indicate the successful completion of the grafting reaction where N-alkylation reactions happen on both sides of the grafting agent 1,4-bis(chloromethyl)benzene. On one side, the reaction takes place with the hydrogen atoms on the imidazole groups of PBI, while on the other side, it reacts with the terminal amino group of PVP. The XPS results provide additional evidence from the perspective of the element. On synthesized PBI-g-PVP surfaces, there is an absence of chlorine (chlorine in grafting agent 1,4-bis(chloromethyl)benzene is substituted) element but a presence of sulfur element (sulfur element in terminal amino PVP appears in PBI), which demonstrates the occurrence of the grafting reaction and PVP is successfully grafted onto PBI. Prepare these modified membranes into MEA. It was found that during the fuel cell operation, all the grafted membranes showed substantial improvement in maximum current density and peak power density compared to unmodified one. For PBI-g-PVP 30, with a grafting degree of 22.4%, the peak power density reaches 1312 mW cm⁻², marking a 59.6% enhancement compared to the pristine PBI membrane. The improvement is caused by the improved PA binding ability of the membrane after grafting. The AST test result shows that the grafting membranes have better long-term durability and performance than unmodified membranes attributed to the presence of added PA binding sites, which can effectively prevent the PA leaching caused by proton migration. In conclusion, the test results indicate that grafting PVP onto PBI is a promising strategy which can effectively improve the fuel cell performance.

Keywords: fuel cell, grafting modification, PA doping ability, PVP

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Authors: Deon Bezuidenhout

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Pre-seeding with autologous endothelial cells improves the long-term patency of synthetic vascular grafts levels obtained with autografts, but is limited to a single centre due to resource, time and other constraints. Spontaneous in vivo endothelialization would obviate the need for pre-seeding, but has been shown to be absent in man due to limited transanastomotic and fallout healing, and the lack of transmural ingrowth due to insufficient porosity. Two types of graft scaffolds with increased interconnected porosity for improved tissue ingrowth and healing are thus proposed and described. Foam-type polyurethane (PU) scaffolds with small, medium and large, interconnected pores were made by phase inversion and spherical porogen extraction, with and without additional surface modification with covalently attached heparin and subsequent loading with and delivery of growth factors. Fibrillar scaffolds were made either by standard electrospinning using degradable PU (Degrapol®), or by dual electrospinning using non-degradable PU. The latter process involves sacrificial fibres that are co-spun with structural fibres and subsequently removed to increased porosity and pore size. Degrapol samples were subjected to in vitro degradation, and all scaffold types were evaluated in vivo for tissue ingrowth and vascularization using rat subcutaneous model. The foam scaffolds were additionally evaluated in a circulatory (rat infrarenal aortic interposition) model that allows for the grafts to be anastomotically and/or ablumenally isolated to discern and determine endothelialization mode. Foam-type grafts with large (150 µm) pores showed improved subcutaneous healing in terms of vascularization and inflammatory response over smaller pore sizes (60 and 90µm), and vascularization of the large porosity scaffolds was significantly increased by more than 70% by heparin modification alone, and by 150% to 400% when combined with growth factors. In the circulatory model, extensive transmural endothelialization (95±10% at 12 w) was achieved. Fallout healing was shown to be sporadic and limited in groups that were ablumenally isolated to prevent transmural ingrowth (16±30% wrapped vs. 80±20% control; p<0.002). Heparinization and GF delivery improved both mural vascularization and lumenal endothelialization. Degrapol electrospun scaffolds showed decrease in molecular mass and corresponding tensile strength over the first 2 weeks, but very little decrease in mass over the 4w test period. Studies on the effect of tissue ingrowth with and without concomitant degradation of the scaffolds, are being used to develop material models for the finite element modelling. In the case of the dual-spun scaffolds, the PU fibre fraction could be controlled shown to vary linearly with porosity (P = −0.18FF +93.5, r2=0.91), which in turn showed inverse linear correlation with tensile strength and elastic modulus (r2 > 0.96). Calculated compliance and burst pressures of the scaffolds increased with fibre fraction, and compliances matching the human popliteal artery (5-10 %/100 mmHg), and high burst pressures (> 2000 mmHg) could be achieved. Increasing porosity (76 to 82 and 90%) resulted in increased tissue ingrowth from 33±7 to 77±20 and 98±1% after 28d. Transmural endothelialization of highly porous foamed grafts is achievable in a circulatory model, and the enhancement of porosity and tissue ingrowth may hold the key the development of spontaneously endothelializing electrospun grafts.

Keywords: electrospinning, endothelialization, porosity, scaffold, vascular graft

Procedia PDF Downloads 268

Authors: Priyanka Mokashi, Aparna Khanna, Nancy Pandita

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Diabetes mellitus is a chronic metabolic disorder which will be the 7th leading cause of death by 2030. The current line of treatment for the diabetes mellitus is oral antidiabetic drugs (biguanides, sulfonylureas, meglitinides, thiazolidinediones and alpha-glycosidase inhibitors) and insulin therapy depending upon the type 1 or type 2 diabetes mellitus. But, these treatments have their disadvantages, ranging from the developing of resistance to the drugs and adverse effects caused by them. Alternative to these synthetic agents, natural products provides a new insight for the development of more efficient and safe drugs due to their therapeutic values. Enicostema littorale blume (A. Raynal) is a traditional Indian plant belongs to the Gentianaceae family. It is widely distributed in Asia, Africa, and South America. There are few reports on Swrtiamarin, major component of this plant for its antidiabetic activity. However, the antidiabetic activity of flavonoids from E. littorale and their mechanism of action have not yet been elucidated. Flavonoids have a positive relationship with disease prevention and can act on various molecular targets and regulate different signaling pathways in pancreatic β-cells, adipocytes, hepatocytes and skeletal myofibers. They may exert beneficial effects in diabetes by (i) improving hyperglycemia through regulation of glucose metabolism in hepatocytes; (ii) enhancing insulin secretion and reducing apoptosis and promoting proliferation of pancreatic β-cells; (iii) increasing glucose uptake in hepatocytes, skeletal muscle and white adipose tissue (iv) reducing insulin resistance, inflammation and oxidative stress. Therefore, we have isolated four flavonoid rich fractions, Fraction A (FA), Fraction B (FB), Fraction C (FC), Fraction D (FD) from crude alcoholic hot (AH) extract from E. littorale, identified by LC/MS. Total eight flavonoids were identified on the basis of fragmentation pattern. Flavonoid FA showed the presence of swertisin, isovitexin, and saponarin; FB showed genkwanin, quercetin, isovitexin, FC showed apigenin, swertisin, quercetin, 5-O-glucosylswertisin and 5-O-glucosylisoswertisin whereas FD showed the presence of swertisin. Further, these fractions were assessed for their antidiabetic activity on stimulating glucose uptake in insulin-resistant HepG2 cell line model (IR/HepG2). The results showed that FD containing C-glycoside Swertisin has significantly increased the glucose uptake rate of IR/HepG2 cells at the concentration of 10 µg/ml as compared to positive control Metformin (0.5mM) which was determined by glucose oxidase- peroxidase method. It has been reported that enhancement of glucose uptake of cells occurs due the translocation of Glut4 vesicles to cell membrane through IR/IRS1/AKT pathway. Therefore, we have studied expressions of three genes IRS1, AKT and Glut4 by real-time PCR to evaluate whether they follow the same pathway or not. It was seen that the glucose uptake rate has increased in FD treated IR/HepG2 cells due to the activation of insulin receptor substrate-1 (IRS1) followed by protein kinase B (AKT) through phosphoinositide 3-kinase (PI3K) leading to translocation of Glut 4 vesicles to cell membrane, thereby enhancing glucose uptake and insulin sensitivity of insulin resistant HepG2 cells. Hence, the up-regulation indicated the mechanism of action through which FD (Swertisin) acts as antidiabetic candidate in the treatment of type 2 diabetes mellitus.

Keywords: E. littorale, glucose transporter, glucose uptake rate, insulin resistance

Procedia PDF Downloads 289

Authors: Mary Ghobrial, Sahar Wefky

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Marine macroalgae have proven to be rich source of bioactive compounds with biomedical potential, not only for human but also for veterinary medicine. Emergence of microbial disease in aquaculture industries implies serious loses. Usage of commercial antibiotics for fish disease treatment produces undesirable side effects. Marine organisms are a rich source of structurally novel biologically active metabolites. Competition for space and nutrients led to the evolution of antimicrobial defense strategies in the aquatic environment. The interest in marine organisms as a potential and promising source of pharmaceutical agents has increased in the last years. Many bioactive and pharmacologically active substances have been isolated from microalgae. Compounds with antibacterial, antifungal and antiviral activities have been also detected in green, brown and red algae. Selected species of marine benthic algae belonging to the Phaeophyta and Rhodophyta, collected from different coastal areas of Alexandria (Egypt), were investigated for their antibacterial and antifungal, activities. Macroalgae samples were collected during low tide from the Alexandria Mediterranean coast. Samples were air dried under shade at room temperature. The dry algae were ground, using electric mixer grinder. They were soaked in 10 ml of each of the solvents acetone, ethanol, methanol and hexane. Antimicrobial activity was evaluated using well-cut diffusion technique In vitro screening of organic solvent extracts from the marine macroalgae Laurencia pinnatifida, Pterocladia capillaceae, Stepopodium zonale, Halopteris scoparia and Sargassum hystrix, showed specific activity in inhibiting the growth of five virulent strains of bacteria pathogenic to fish Pseudomonas fluorescens, Aeromonas hydrophila, Vibrio anguillarum, V. tandara, Escherichia coli and two fungi Aspergillus flavus and A. niger. Results showed that, acetone and ethanol extracts of all test macroalgae exhibited antibacterial activity, while acetone extract of the brown Sargassum hystrix displayed the highest antifungal activity. The extracts of seaweeds inhibited bacteria more strongly than fungi and species of the Rhodophyta showed the greatest activity against the bacteria rather than fungi tested. The gas liquid chromatography coupled with mass spectrometry detection technique allows good qualitative and quantitative analysis of the fractionated extracts with high sensitivity to the smaller amounts of components. Results indicated that, the main common component in the acetone extracts of L. pinnatifida and P. capillacea is 4-hydroxy-4-methyl2-pentanone representing 64.38 and 58.60%. Thus, the extracts derived from the red macroalgae were more efficient than those obtained from the brown macroalgae in combating bacterial pathogens rather than pathogenic fungi. The most preferred species over all was the red Laurencia pinnatifida. In conclusion, the present study provides the potential of red and brown macroalgae extracts for development of anti-pathogenic agents for use in fish aquaculture.

Keywords: bacteria, fungi, extracts, solvents

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Authors: R. Di Lorenzo, S. Laneri, A. Sacchi

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Hair surface properties affect hair texture and shine, whereas the healthy state of the hair cortex sways hair ends. Even if cosmetic treatments are intrinsically safe, there is potentially damaging action on the hair fibers. Loss of luster, frizz, split ends, and other hair problems are particularly prevalent among people who repeatedly alter the natural style of their hair or among people with intrinsically weak hair. Technological and scientific innovations in hair care thus become invaluable allies to preserve their natural well-being and shine. The study evaluated restoring keratin-like ingredients that improve hair fibers' structural integrity, increase tensile strength, improve hair manageability and moisturizing. The hair shaft is composed of 65 - 95% of keratin. It gives the hair resistance, elasticity, and plastic properties and also contributes to their waterproofing. Providing exogenous keratin is, therefore, a practical approach to protect and nourish the hair. By analyzing the amino acid composition of keratin, we find a high frequency of hydrophobic amino acids. It confirms the critical role interactions, mainly hydrophobic, between cosmetic products and hair. The active ingredient analyzed comes from vegetable proteins through an enzymatic cut process that selected only oligo- and polypeptides (> 3500 KDa) rich in amino acids with hydrocarbon side chains apolar or sulfur. These chemical components are the most expressed amino acids at the level of the capillary keratin structure, and it determines the most significant possible compatibility with the target substrate. Given the biological variability of the sources, it isn't easy to define a constant and reproducible molecular formula of the product. Still, it consists of hydroxypropiltrimonium vegetable peptides with keratin-like performances. 20 natural hair tresses (30 cm in length and 0.50 g weight) were treated with the investigated products (5 % v/v aqueous solution) following a specific protocol and compared with non-treated (Control) and benchmark-keratin-treated strands (Benchmark). Their brightness, moisture content, cortical and surface integrity, and tensile strength were evaluated and statistically compared. Keratin-like treated hair tresses showed better results than the other two groups (Control and Benchmark). The product improves the surface with significant regularization of the cuticle closure, improves the cortex and the peri-medullar area filling, gives a highly organized and tidy structure, delivers a significant amount of sulfur on the hair, and is more efficient moisturization and imbibition power, increases hair brightness. The hydroxypropyltrimonium quaternized group added to the C-terminal end interacts with the negative charges that form on the hair after washing when disheveled and tangled. The interactions anchor the product to the hair surface, keeping the cuticles adhered to the shaft. The small size allows the peptides to penetrate and give body to the hair, together with a conditioning effect that gives an image of healthy hair. Results suggest that the product is a valid ally in numerous restructuring/conditioning, shaft protection, straightener/dryer-damage prevention hair care product.

Keywords: conditioning, hair damage, hair, keratin, polarized light microscopy, scanning electron microscope, thermogravimetric analysis

Procedia PDF Downloads 105

Authors: S. J. Addinell, A. F. Grabsch, P. D. Fawell, B. Evans

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The Deep Exploration Technologies Cooperative Research Centre (DET CRC) is researching and developing a new coiled tubing based greenfields mineral exploration drilling system utilising down-hole water-powered percussive drill tooling. This new drilling system is aimed at significantly reducing the costs associated with identifying mineral resource deposits beneath deep, barren cover. This system has shown superior rates of penetration in water-rich, hard rock formations at depths exceeding 500 metres. With fluid flow rates of up to 120 litres per minute at 200 bar operating pressure to energise the bottom hole tooling, excessive quantities of high quality drilling fluid (water) would be required for a prolonged drilling campaign. As a result, drilling fluid recovery and recycling has been identified as a necessary option to minimise costs and logistical effort. While the majority of the cuttings report as coarse particles, a significant fines fraction will typically also be present. To maximise tool life longevity, the percussive bottom hole assembly requires high quality fluid with minimal solids loading and any recycled fluid needs to have a solids cut point below 40 microns and a concentration less than 400 ppm before it can be used to reenergise the system. This paper presents experimental results obtained from the research program during laboratory and field testing of the prototype drilling system. A study of the morphological aspects of the cuttings generated during the percussive drilling process shows a strong power law relationship for particle size distributions. This data is critical in optimising solids control strategies and cuttings dewatering techniques. Optimisation of deployable solids control equipment is discussed and how the required centrate clarity was achieved in the presence of pyrite-rich metasediment cuttings. Key results were the successful pre-aggregation of fines through the selection and use of high molecular weight anionic polyacrylamide flocculants and the techniques developed for optimal dosing prior to scroll decanter centrifugation, thus keeping sub 40 micron solids loading within prescribed limits. Experiments on maximising fines capture in the presence of thixotropic drilling fluid additives (e.g. Xanthan gum and other biopolymers) are also discussed. As no core is produced during the drilling process, it is intended that the particle laden returned drilling fluid is used for top-of-hole geochemical and mineralogical assessment. A discussion is therefore presented on the biasing and latency of cuttings representivity by dewatering techniques, as well as the resulting detrimental effects on depth fidelity and accuracy. Data pertaining to the sample biasing with respect to geochemical signatures due to particle size distributions is presented and shows that, depending on the solids control and dewatering techniques used, it can have unwanted influence on top-of-hole analysis. Strategies are proposed to overcome these effects, improving sample quality. Successful solids control and cuttings dewatering for water-powered percussive drilling is presented, contributing towards the successful advancement of coiled tubing based greenfields mineral exploration.

Keywords: cuttings, dewatering, flocculation, percussive drilling, solids control

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Authors: Omaima A. Sharaf, Wafaa M. Abd El-Rahim, Hassan Moawad, Michael J. Sadowsky

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Textile industry is one of the important industries in the worldwide. It is known that the eco-friendly industrial and agricultural activities are significant for socio-economic stability of all countries. The absence of appropriate industrial waste water treatments is essential barrier for sustainable development in food and agricultural sectors especially in developing country like Egypt. Thus, the development of enzymatic bioremediation technology for textile dye removal will enhance the collaboration between scientists who develop the technology and industry where this technology will be implemented towards the safe disposal of the textile dye wastes. Highly efficient microorganisms are of most importance in developing and using highly effective biological treatment processes. Bacterial degradation of azo dyes is generally initiated by an enzymatic step that involves cleavage of azo linkages, usually with the aid of an azoreductase as electron donor. Thus, expanding the spectrum of microorganisms with high enzymatic activities as azoreductases and discovering novel azo-dye degrading enzymes, with enhanced stability and superior catalytic properties, are necessary for many environmental and industrial applications. Consequently, the use of molecular tools has become increasingly integrated into the understanding of enzyme properties and characterization. Researchers have utilized a gene cloning and expression methods as a tool to produce recombinant protein for decolorizing dyes more efficiently. Thus, presumptive evidence for the presence of genes encoding azoreductases in the genomes of selected local, and most potent azo-degrading strains were obtained by using specific oligonucleotides primers. These potent strains have been isolated from textile industrial wastewater in Egypt and identified using 16S rRNA sequence analysis as 'Lysinibacillus macrolidesB8, Brevibacillus parabrevisB11, Bacillus coagulansB7, and B. cereusB5'. PCR products of two full length genes designated as (AZO1;621bp and AZO2;534bp) were detected. BLASTx results indicated that AZO1 gene was corresponding to predicted azoreductase from of Bacillus sp. ABP14, complete genome, multispecies azoreductase [Bacillus], It was submitted to the gene bank by an accession no., BankIt2085371 AZO1 MG923210 (621bp; 207 amino acids). AZO1 was generated from the DNA of our identified strains Lysinibacillus macrolidesB8. On the other hand, AZO2 gene was corresponding to a predicted azoreductase from Bacillus cereus strain S2-8. Gene bank accession no. was BankIt2085839 AZO2 MG932081 (534bp;178 amino acids) and it was amplified from our Bacillus coagulansB7. Both genes were successfully cloned into pCR2.1TOPO (Invitrogen) and in pET28b+ vectors, then they transformed into E. coli DH5α and BL21(DE3) cells for heterologous expression studies. Our recombinant azoreductases (AZO1&AZO2) exhibited potential enzyme activity and efficiently decolorized an azo dye (Direct violet). They exhibited pH stability between 6 and 8 with optimum temperature up to 60°C and 37 °C after induction by 1mM and 1.5mM IPTG, for both AZO1 &AZO2, respectively. These results suggested that further optimization and purification of these recombinant proteins by using different heterologous expression systems will give great potential for the sustainable utilization of these recombinant enzymes in several industrial applications especially in wastewater treatments.

Keywords: azoreductases, decolorization, enzyme activity, gene cloning and expression

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Authors: Olga V. Chervyakova, Elmira T. Tailakova, Vitaliy M. Strochkov, Kulyaisan T. Sultankulova, Nurlan T. Sandybayev, Lev G. Nemchinov, Rosemarie W. Hammond

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Sheep pox is a highly contagious infection that OIE regards to be one of the most dangerous animal diseases. It causes enormous economic losses because of death and slaughter of infected animals, lower productivity, cost of veterinary and sanitary as well as quarantine measures. To control spread of sheep pox infection the attenuated vaccines are widely used in the Republic of Kazakhstan and other Former Soviet Union countries. In spite of high efficiency of live vaccines, the possible presence of the residual virulence, potential genetic instability restricts their use in disease-free areas that leads to necessity to exploit new approaches in vaccine development involving recombinant DNA technology. Vaccines on the basis of recombinant proteins are the newest generation of prophylactic preparations. The main advantage of these vaccines is their low reactogenicity and this fact makes them widely used in medical and veterinary practice for vaccination of humans and farm animals. The objective of the study is to produce recombinant immunogenic proteins for development of the high-performance means for sheep pox prophylaxis. The SPV proteins were chosen for their homology with the known immunogenic vaccinia virus proteins. Assay of nucleotide and amino acid sequences of the target SPV protein genes. It has been shown that four proteins SPPV060 (ortholog L1), SPPV074 (ortholog H3), SPPV122 (ortholog A33) and SPPV141 (ortholog B5) possess transmembrane domains at N- or C-terminus while in amino acid sequences of SPPV095 (ortholog А 4) and SPPV117 (ortholog А 27) proteins these domains were absent. On the basis of these findings the primers were constructed. Target genes were amplified and subsequently cloned into the expression vector рЕТ26b(+) or рЕТ28b(+). Six constructions (pSPPV060ΔТМ, pSPPV074ΔТМ, pSPPV095, pSPPV117, pSPPV122ΔТМ and pSPPV141ΔТМ) were obtained for expression of the SPV genes under control of T7 promoter in Escherichia coli. To purify and detect recombinant proteins the amino acid sequences were modified by adding six histidine molecules at C-terminus. Induction of gene expression by IPTG was resulted in production of the proteins with molecular weights corresponding to the estimated values for SPPV060, SPPV074, SPPV095, SPPV117, SPPV122 and SPPV141, i.e. 22, 30, 20, 19, 17 and 22 kDa respectively. Optimal protocol of expression for each gene that ensures high yield of the recombinant protein was identified. Assay of cellular lysates by western blotting confirmed expression of the target proteins. Recombinant proteins bind specifically with antibodies to polyhistidine. Moreover all produced proteins are specifically recognized by the serum from experimentally SPV-infected sheep. The recombinant proteins SPPV060, SPPV074, SPPV117, SPPV122 and SPPV141 were also shown to induce formation of antibodies with virus-neutralizing activity. The results of the research will help to develop a new-generation high-performance means for specific sheep pox prophylaxis that is one of key moments in animal health protection. The research was conducted under the International project ISTC # K-1704 “Development of methods to construct recombinant prophylactic means for sheep pox with use of transgenic plants” and under the Grant Project RK MES G.2015/0115RK01983 "Recombinant vaccine for sheep pox prophylaxis".

Keywords: prophylactic preparation, recombinant protein, sheep pox virus, subunit vaccine

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Authors: Liang Zhao, Chongquan Zhong

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Introduction: According to statistics, people spend 80% to 90% of time indoor, so indoor air quality, either at home or in the office, greatly impacts the quality of life, health and work efficiency. Therefore, indoor air quality is very important to human activities. With the acceleration of urbanization, people are spending more time in indoor activity. The time in indoor environment, the living space, and the frequency interior decoration are all increasingly increased. However, housing decoration materials contain formaldehyde and other harmful substances, causing environmental and air quality problems, which have brought serious damage to countless families and attracted growing attention. According to World Health Organization statistics, the indoor environments in more than 30% of buildings in China are polluted by poisonous and harmful gases. Indoor pollution has caused various health problems, and these widespread public health problems can lead to respiratory diseases. Long-term inhalation of low-concentration formaldehyde would cause persistent headache, insomnia, weakness, palpitation, weight loss and vomiting, which are serious impacts on human health and safety. On the other hand, as for offices, some surveys show that good indoor air quality helps to enthuse the staff and improve the work efficiency by 2%-16%. Therefore, people need to further understand the living and working environments. There is a need for easy-to-use indoor environment monitoring instruments, with which users only have to power up and monitor the environmental parameters. The corresponding real-time data can be displayed on the screen for analysis. Environment monitoring should have the sensitive signal alarm function and send alarm when harmful gases such as formaldehyde, CO, SO2, are excessive to human body. System design: According to the monitoring requirements of various gases, temperature and humidity, we designed a portable, light, real-time and accurate monitor for various environmental parameters, including temperature, humidity, formaldehyde, methane, and CO. This monitor will generate an alarm signal when a target is beyond the standard. It can conveniently measure a variety of harmful gases and provide the alarm function. It also has the advantages of small volume, convenience to carry and use. It has a real-time display function, outputting the parameters on the LCD screen, and a real-time alarm function. Conclusions: This study is focused on the research and development of a portable parameter monitoring instrument for indoor environment. On the platform of an STM32 development board, the monitored data are collected through an external sensor. The STM32 platform is for data acquisition and processing procedures, and successfully monitors the real-time temperature, humidity, formaldehyde, CO, methane and other environmental parameters. Real-time data are displayed on the LCD screen. The system is stable and can be used in different indoor places such as family, hospital, and office. Meanwhile, the system adopts the idea of modular design and is superior in transplanting. The scheme is slightly modified and can be used similarly as the function of a monitoring system. This monitor has very high research and application values.

Keywords: indoor air quality, gas concentration detection, embedded system, sensor

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Authors: Alexis Grandcoin, Stephanie Piel, Estelle Baures

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Brittany (France) is an agricultural region, where emerging pollutants are highly at risk to reach water bodies. Among them, pesticides metabolites are frequently detected in surface waters. The Vilaine watershed (11 000 km²) is of great interest, as a large drinking water treatment plant (100 000 m³/day) is located at the extreme downstream of it. This study aims to provide an evaluation of the pesticides metabolites pollution in the Vilaine watershed, and an understanding of their availability, in order to protect the water resource. Hydroclimatic conditions, land use, and water quality parameters controlling metabolites availability are emphasized. Later this knowledge will be used to understand the favoring conditions resulting in metabolites export towards surface water. 19 sampling points have been strategically chosen along the 220 km of the Vilaine river and its 3 main influents. Furthermore, the intakes of two drinking water plants have been sampled, one is located at the extreme downstream of the Vilaine river and the other is the riparian groundwater under the Vilaine river. 5 sampling campaigns with various hydroclimatic conditions have been carried out. Water quality parameters and hydroclimatic conditions have been measured. 15 environmentally relevant pesticides and metabolites have been analyzed. Also, these compounds are recalcitrant to classic water treatment that is why they have been selected. An evaluation of the watershed contamination has been done in 2016-2017. First observations showed that aminomethylphosphonic acid (AMPA) and metolachlor ethanesulfonic acid (MESA) are the most detected compounds in surface waters samples with 100 % and 98 % frequency of detection respectively. They are the main pollutants of the watershed regardless of the hydroclimatic conditions. AMPA concentration in the river strongly increases downstream of Rennes agglomeration (220k inhabitants) and reaches a maximum of 2.3 µg/l in low waters conditions. Groundwater contains mainly MESA, Diuron and metazachlor ESA at concentrations close to limits of quantification (LOQ) (0.02 µg/L). Metolachlor, metazachlor and alachlor due to their fast degradation in soils were found in small amounts (LOQ – 0.2 µg/L). Conversely glyphosate was regularly found during warm and sunny periods up to 0.6 µg/L. Soil uses (agricultural cultures types, urban areas, forests, wastewater treatment plants implementation), water quality parameters, and hydroclimatic conditions have been correlated to pesticides and metabolites concentration in waters. Statistical treatments showed that chloroacetamides metabolites and AMPA behave differently regardless of the hydroclimatic conditions. Chloroacetamides are correlated to each other, to agricultural areas and to typical agricultural tracers as nitrates. They are present in waters the whole year, especially during rainy periods, suggesting important stocks in soils. Also Chloroacetamides are negatively correlated with AMPA, the different forms of phosphorus, and organic matter. AMPA is ubiquitous but strongly correlated with urban areas despite the recent French regulation, restricting glyphosate to agricultural and private uses. This work helps to predict and understand metabolites present in the water resource used to craft drinking water. As the studied metabolites are difficult to remove, this project will be completed by a water treatment part.

Keywords: agricultural watershed, AMPA, metolachlor-ESA, water resource

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Authors: Marta Paszkiewicz, Justyna Łuczak, Martyna Marchelek, Adriana Zaleska-Medynska

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In recent years, photocatalytical processes have been intensively investigated for destruction of pollutants, hydrogen evolution, disinfection of water, air and surfaces, for the construction of self-cleaning materials (tiles, glass, fibres, etc.). Titanium dioxide (TiO2) is the most popular material used in heterogeneous photocatalysis due to its excellent properties, such as high stability, chemical inertness, non-toxicity and low cost. It is well known that morphology and microstructure of TiO2 significantly influence the photocatalytic activity. This characteristics as well as other physical and structural properties of photocatalysts, i.e., specific surface area or density of crystalline defects, could be controlled by preparation route. In this regard, TiO2 particles can be obtained by sol-gel, hydrothermal, sonochemical methods, chemical vapour deposition and alternatively, by ionothermal synthesis using ionic liquids (ILs). In the TiO2 particles synthesis ILs may play a role of a solvent, soft template, reagent, agent promoting reduction of the precursor or particles stabilizer during synthesis of inorganic materials. In this work, the effect of the ILs anion type on morphology and photoactivity of TiO2 is presented. The preparation of TiO2 microparticles with spherical structure was successfully achieved by solvothermal method, using tetra-tert-butyl orthotitatane (TBOT) as the precursor. The reaction process was assisted by an ionic liquids 1-butyl-3-methylimidazolium bromide [BMIM][Br], 1-butyl-3-methylimidazolium tetrafluoroborate [BMIM][BF4] and 1-butyl-3-methylimidazolium haxafluorophosphate [BMIM][PF6]. Various molar ratios of all ILs to TBOT (IL:TBOT) were chosen. For comparison, reference TiO2 was prepared using the same method without IL addition. Scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), Brenauer-Emmett-Teller surface area (BET), NCHS analysis, and FTIR spectroscopy were used to characterize the surface properties of the samples. The photocatalytic activity was investigated by means of phenol photodegradation in the aqueous phase as a model pollutant, as well as formation of hydroxyl radicals based on detection of fluorescent product of coumarine hydroxylation. The analysis results showed that the TiO2 microspheres had spherical structure with the diameters ranging from 1 to 6 µm. The TEM micrographs gave a bright observation of the samples in which the particles were comprised of inter-aggregated crystals. It could be also observed that the IL-assisted TiO2 microspheres are not hollow, which provides additional information about possible formation mechanism. Application of the ILs results in rise of the photocatalytic activity as well as BET surface area of TiO2 as compared to pure TiO2. The results of the formation of 7-hydroxycoumarin indicated that the increased amount of ·OH produced at the surface of excited TiO2 for samples TiO2_ILs well correlated with more efficient degradation of phenol. NCHS analysis showed that ionic liquids remained on the TiO2 surface confirming structure directing role of that compounds.

Keywords: heterogeneous photocatalysis, IL-assisted synthesis, ionic liquids, TiO2

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Authors: Gabrielle Robertson, Matthew Downs, Joseph Dagher

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Iron deposition in the brain has been linked with a host of neurological disorders such as Alzheimer’s, Parkinson’s, and Multiple Sclerosis. While some treatment options exist, there are no objective measurement tools that allow for the monitoring of iron levels in the brain in vivo. An emerging Magnetic Resonance Imaging (MRI) method has been recently proposed to deduce iron concentration through quantitative measurement of magnetic susceptibility. This is a multi-step process that involves repeated modeling of physical processes via approximate numerical solutions. For example, the last two steps of this Quantitative Susceptibility Mapping (QSM) method involve I) mapping magnetic field into magnetic susceptibility and II) mapping magnetic susceptibility into iron concentration. Process I involves solving an ill-posed inverse problem by using regularization via injection of prior belief. The end result from Process II highly depends on the model used to describe the molecular content of each voxel (type of iron, water fraction, etc.) Due to these factors, the accuracy and repeatability of QSM have been an active area of research in the MRI and medical imaging community. This work aims to estimate iron concentration in the brain via a single step. A synthetic numerical model of the human head was created by automatically and manually segmenting the human head on a high-resolution grid (640x640x640, 0.4mm³) yielding detailed structures such as microvasculature and subcortical regions as well as bone, soft tissue, Cerebral Spinal Fluid, sinuses, arteries, and eyes. Each segmented region was then assigned tissue properties such as relaxation rates, proton density, electromagnetic tissue properties and iron concentration. These tissue property values were randomly selected from a Probability Distribution Function derived from a thorough literature review. In addition to having unique tissue property values, different synthetic head realizations also possess unique structural geometry created by morphing the boundary regions of different areas within normal physical constraints. This model of the human brain is then used to create synthetic MRI measurements. This is repeated thousands of times, for different head shapes, volume, tissue properties and noise realizations. Collectively, this constitutes a training-set that is similar to in vivo data, but larger than datasets available from clinical measurements. This 3D convolutional U-Net neural network architecture was used to train data-driven Deep Learning models to solve for iron concentrations from raw MRI measurements. The performance was then tested on both synthetic data not used in training as well as real in vivo data. Results showed that the model trained on synthetic MRI measurements is able to directly learn iron concentrations in areas of interest more effectively than other existing QSM reconstruction methods. For comparison, models trained on random geometric shapes (as proposed in the Deep QSM method) are less effective than models trained on realistic synthetic head models. Such an accurate method for the quantitative measurement of iron deposits in the brain would be of important value in clinical studies aiming to understand the role of iron in neurological disease.

Keywords: magnetic resonance imaging, MRI, iron deposition, machine learning, quantitative susceptibility mapping

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Authors: Thanthrige Thiunuwan Priyathilaka, Don Anushka Sandaruwan Elvitigala, Bong-Soo Lim, Hyung-Bok Jeong, Jehee Lee

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Toll like receptors (TLRs) are a phylogeneticaly conserved family of pattern recognition receptors, which participates in the host immune responses against various pathogens and pathogen derived mitogen. TLR21, a non-mammalian type, is almost restricted to the fish species even though those can be identified rarely in avians and amphibians. Herein, this study was carried out to identify and characterize TLR21 from rock bream (Oplegnathus fasciatus) designated as RbTLR21, at transcriptional and genomic level. In this study, the full length cDNA and genomic sequence of RbTLR21 was identified using previously constructed cDNA sequence database and BAC library, respectively. Identified RbTLR21 sequence was characterized using several bioinformatics tools. The quantitative real time PCR (qPCR) experiment was conducted to determine tissue specific expressional distribution of RbTLR21. Further, transcriptional modulation of RbTLR21 upon the stimulation with Streptococcus iniae (S. iniae), rock bream iridovirus (RBIV) and Edwardsiella tarda (E. tarda) was analyzed in spleen tissues. The complete coding sequence of RbTLR21 was 2919 bp in length which can encode a protein consisting of 973 amino acid residues with molecular mass of 112 kDa and theoretical isoelectric point of 8.6. The anticipated protein sequence resembled a typical TLR domain architecture including C-terminal ectodomain with 16 leucine rich repeats, a transmembrane domain, cytoplasmic TIR domain and signal peptide with 23 amino acid residues. Moreover, protein folding pattern prediction of RbTLR21 exhibited well-structured and folded ectodomain, transmembrane domain and cytoplasmc TIR domain. According to the pair wise sequence analysis data, RbTLR21 showed closest homology with orange-spotted grouper (Epinephelus coioides) TLR21with 76.9% amino acid identity. Furthermore, our phylogenetic analysis revealed that RbTLR21 shows a close evolutionary relationship with its ortholog from Danio rerio. Genomic structure of RbTLR21 consisted of single exon similar to its ortholog of zebra fish. Sevaral putative transcription factor binding sites were also identified in 5ʹ flanking region of RbTLR21. The RBTLR 21 was ubiquitously expressed in all the tissues we tested. Relatively, high expression levels were found in spleen, liver and blood tissues. Upon induction with rock bream iridovirus, RbTLR21 expression was upregulated at the early phase of post induction period even though RbTLR21 expression level was fluctuated at the latter phase of post induction period. Post Edwardsiella tarda injection, RbTLR transcripts were upregulated throughout the experiment. Similarly, Streptococcus iniae induction exhibited significant upregulations of RbTLR21 mRNA expression in the spleen tissues. Collectively, our findings suggest that RbTLR21 is indeed a homolog of TLR21 family members and RbTLR21 may be involved in host immune responses against bacterial and DNA viral infections.

Keywords: rock bream, toll like receptor 21 (TLR21), pattern recognition receptor, genomic characterization

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Authors: Ammar Alali, Mahmoud Abughaban

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Cost reduction and drilling optimization is the goal of many drilling operators. Historically, stuck pipe incidents were a major segment of non-productive time (NPT) associated costs. Traditionally, stuck pipe problems are part of the operations and solved post-sticking. However, the real key to savings and success is in predicting the stuck pipe incidents and avoiding the conditions leading to its occurrences. Previous attempts in stuck-pipe predictions have neglected the local geology of the problem. The proposed predictive tool utilizes geophysical data processing techniques and Machine Learning (ML) algorithms to predict drilling activities events in real-time using surface drilling data with minimum computational power. The method combines two types of analysis: (1) real-time prediction, and (2) cause analysis. Real-time prediction aggregates the input data, including historical drilling surface data, geological formation tops, and petrophysical data, from wells within the same field. The input data are then flattened per the geological formation and stacked per stuck-pipe incidents. The algorithm uses two physical methods (stacking and flattening) to filter any noise in the signature and create a robust pre-determined pilot that adheres to the local geology. Once the drilling operation starts, the Wellsite Information Transfer Standard Markup Language (WITSML) live surface data are fed into a matrix and aggregated in a similar frequency as the pre-determined signature. Then, the matrix is correlated with the pre-determined stuck-pipe signature for this field, in real-time. The correlation used is a machine learning Correlation-based Feature Selection (CFS) algorithm, which selects relevant features from the class and identifying redundant features. The correlation output is interpreted as a probability curve of stuck pipe incidents prediction in real-time. Once this probability passes a fixed-threshold defined by the user, the other component, cause analysis, alerts the user of the expected incident based on set pre-determined signatures. A set of recommendations will be provided to reduce the associated risk. The validation process involved feeding of historical drilling data as live-stream, mimicking actual drilling conditions, of an onshore oil field. Pre-determined signatures were created for three problematic geological formations in this field prior. Three wells were processed as case studies, and the stuck-pipe incidents were predicted successfully, with an accuracy of 76%. This accuracy of detection could have resulted in around 50% reduction in NPT, equivalent to 9% cost saving in comparison with offset wells. The prediction of stuck pipe problem requires a method to capture geological, geophysical and drilling data, and recognize the indicators of this issue at a field and geological formation level. This paper illustrates the efficiency and the robustness of the proposed cross-disciplinary approach in its ability to produce such signatures and predicting this NPT event.

Keywords: drilling optimization, hazard prediction, machine learning, stuck pipe

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