958 resultados para Associated production
Resumo:
We investigate the ways young children’s use of mobile touchscreen interfaces is both understood and shaped by parents through the production of YouTube videos and discussions in associated comment threads. This analysis expands on, and departs from, theories of parental mediation, which have traditionally been framed through a media effects approach in analyzing how parents regulate their children’s use of broadcast media, such as television, within family life. We move beyond the limitations of an effects framing through more culturally and materially oriented theoretical lenses of mediation, considering the role mobile interfaces now play in the lives of infants through analysis of the ways parents intermediate between domestic spaces and networked publics. We propose the concept of intermediation, which builds on insights from critical interface studies as well as cultural industries literature to help account for these expanded aspects of digital parenting. Here, parents are not simply moderating children’s media use within the home, but instead operating as an intermediary in contributing to online representations and discourses of children’s digital culture. This intermediary role of parents engages with ideological tensions in locating notions of “naturalness:” the iPad’s gestural interface or the child’s digital dexterity.
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Adeno-associated viral (AAV) vectors are among the most widely used gene transfer systems in basic and pre-clinical research and have been employed in more than 160 clinical trials. AAV vectors are commonly produced in producer cell lines like HEK293 by co-transfection with a so-called vector plasmid and one (in this work) or two so-called helper plasmids. The vector plasmid contains the transgene cassette of interest (TEC) flanked by AAV’s inverted terminal repeats (ITRs) which serve as packaging signals, whereas the helper plasmid provides the required AAV and helper virus functions in trans. A pivotal aspect of AAV vectorology is the manufacturing of AAV vectors free from impurities arising during the production process. These impurities include AAV vector preparations that contain capsids containing prokaryotic sequences, e.g. antibiotic resistance genes originating from the producer plasmids. In the first part of the thesis we aimed at improving the safety of AAV vectors. As we found that encapsidated prokaryotic sequences (using the ampicillin resistance gene as indicator) cannot be re-moved by standard purification methods we investigated whether the producer plasmids could be replaced by Minicircles (MCs). MCs are circular DNA constructs which contain no functional or coding prokaryotic sequences; they only consist of the TEC and a short sequence required for production and purification. MC counterparts of a vector plasmid encoding for enhanced green fluorescent (eGFP) protein and a helper plasmid encoding for AAV serotype 2 (AAV2) and helper Adenovirus (Ad) genes were designed and produced by PlasmidFactory (Bielefeld, Germany). Using all four possible combinations of plasmid and MCs, single-stranded AAV2 vectors (ssAAV) and self-complementary AAV vectors (scAAV) were produced and characterized for vector quantity, quality and functionality. The analyses showed that plasmids can be replaced by MCs without decreasing the efficiency of vector production and vector quality. MC-derived scAAV vector preparations even exceeded plasmid-derived preparations, as they displayed up to 30-fold improved transduction efficiencies. Using MCs as tools, we found that the vector plasmid is the main source of encapsidated prokaryotic sequences. Remarkably, we found that plasmid-derived scAAV vector preparations contained a much higher relative amount of prokaryotic sequences (up to 26.1 %, relative to TEC) compared to ssAAV vector preparations (up to 2.9 %). By replacing both plasmids by MCs the amount of functional prokaryotic sequences could be decreased to below the limit of quantification. Additional analyses for DNA impurities other than prokaryotic sequences showed that scAAV vectors generally contained a higher amount of non-vector DNA (e.g. adenoviral sequences) than ssAAV vectors. For both, ssAAV and scAAV vector preparations, MC-derived vectors tended to contain lower amounts of foreign DNA. None of the vectors tested could be shown to induce immunogenicity. In summary we could demonstrate that the quality of AAV vector preparations could be significantly improved by replacing producer plasmids by MCs. Upon transduction of a target tissue, AAV vector genomes predominantly remain in an episomal state, as duplex DNA circles or concatemers. These episomal forms mediate long-term transgene expression in terminally differentiated cells, but are lost in proliferating cells due to cell division. Therefore, in the second part of the thesis, in cooperation with Claudia Hagedorn and Hans J. Lipps (University Witten/Herdecke) an AAV vector genome was equipped with an autonomous replication element (Scaffold/matrix attachment region (S/MAR)). AAV-S/MAR encoding for eGFP and a blasticidin resistance gene and a control vector with the same TEC but lacking the S/MAR element (AAV-ΔS/MAR) were produced and transduced into highly proliferative HeLa cells. Antibiotic pressure was employed to select for cells stably maintaining the vector genome. AAV-S/MAR transduced cells yielded a higher number of colonies than AAV-ΔS/MAR-transduced cells. Colonies derived from each vector transduction were picked and cultured further. They remained eGFP-positive (up to 70 days, maximum cultivation period) even in the absence of antibiotic selection pressure. Interestingly, the mitotic stability of both AAV-S/MAR and control vector AAV-ΔS/MAR was found to be a result of episomal maintenance of the vector genome. This finding indicates that, under specific conditions such as the mild selection pressure we employed, “common” AAV vectors persist episomally. Thus, the S/MAR element increases the establishment frequency of stable episomes, but is not a prerequisite.
Resumo:
Beef production can be environmentally detrimental due in large part to associated enteric methane (CH4) production, which contributes to climate change. However, beef production in well-managed grazing systems can aid in soil carbon sequestration (SCS), which is often ignored when assessing beef production impacts on climate change. To estimate the carbon footprint and climate change mitigation potential of upper Midwest grass-finished beef production systems, we conducted a partial life cycle assessment (LCA) comparing two grazing management strategies: 1) a non-irrigated, lightly-stocked (1.0 AU/ha), high-density (100,000 kg LW/ha) system (MOB) and 2) an irrigated, heavily-stocked (2.5 AU/ha), low-density (30,000 kg LW/ha) system (IRG). In each system, April-born steers were weaned in November, winter-backgrounded for 6 months and grazed until their endpoint the following November, with average slaughter age of 19 months and a 295 kg hot carcass weight. As the basis for the LCA, we used two years of data from Lake City Research Center, Lake City, MI. We included greenhouse gas (GHG) emissions associated with enteric CH4, soil N2O and CH4 fluxes, alfalfa and mineral supplementation, and farm energy use. We also generated results from the LCA using the enteric emissions equations of the Intergovernmental Panel on Climate Change (IPCC). We evaluated a range of potential rates of soil carbon (C) loss or gain of up to 3 Mg C ha-1 yr-1. Enteric CH4 had the largest impact on total emissions, but this varied by grazing system. Enteric CH4 composed 62 and 66% of emissions for IRG and MOB, respectively, on a land basis. Both MOB and IRG were net GHG sources when SCS was not considered. Our partial LCA indicated that when SCS potential was included, each grazing strategy could be an overall sink. Sensitivity analyses indicated that soil in the MOB and IRG systems would need to sequester 1 and 2 Mg C ha-1 yr-1 for a net zero GHG footprint, respectively. IPCC model estimates for enteric CH4 were similar to field estimates for the MOB system, but were higher for the IRG system, suggesting that 0.62 Mg C ha-1 yr-1 greater SCS would be needed to offset the animal emissions in this case.
Resumo:
Greenhouse production is a very important activity in the West region of Portugal, with an area of approximately 800 ha where the regular production consists in two crops per year, one in winter-spring and the other in summer-autumn. Many growers are now prepared to better exploit market opportunities, since they know that the big export window opportunity is from June to September, when the production is difficult in other regions of south due to high temperatures. Grower’s use new and more productive varieties, either in soil or hydroponic systems, mostly in unheated greenhouses, naturally ventilated, and equipped with modern fertigation systems. Greenhouse production causes some environmental impacts due to the high use of inputs. Several improvements in technologies and crop practices may contribute to increase the use efficiency of resources, decreasing the negative environmental impacts. Greenhouse vegetable production in Northern EU countries is based on the supply of heating and differs significantly from the production system in the Southern EU countries. In the Northern countries, direct energy inputs, mostly for heating, are predominant while in the South the indirect energy input is also important, mainly associated with fertilizers, plastic cover materials and other auxiliary materials. The main objective of this work was to characterise the greenhouse production systems in the West region of Portugal, in order to evaluate the energetic consumptions (direct and indirect), the GHH emissions, the production costs and the farmer’s income. With this work the mostly important inputs were identified, allowing proposing alternative measures to improve efficiency and sustainability. All the data was obtained by surveys performed directly with growers, previously selected to be representative of the crop practices and greenhouse type of the region. However, more research should be performed in order to develop and to test technologies capable to improve resources use efficiency in greenhouse production.
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Fungal endophytes present in different asymptomatic grapevine plants (Vitis vinifera L.) located in different vineyards within Alentejo, a highly important viticulture region in Portugal, were identified in this study. Sampled grapevine plants included the three most representative cultivars in the region, Syrah, Cabernet Sauvignon, and Aragonez, growing under two different modes of management, conventional and biological. Sixteen fungal taxa were identified through sequencing of the internal transcribed spacer region. Total number of endophytic fungi isolated showed significant differences both in management mode and in cultivars, with higher numbers in grapevines under conventional mode and from Syrah cultivar. The composition of fungal endophytic communities did not show significant differences among cultivars, but differences were observed between fungal communities isolated from grapevines under biological or conventional modes. The most fungal taxa isolated from grapevines cultivated under biological mode were Alternaria alternata, Cladosporium sp., and Nigrospora oryzae, and under conventional mode Botrytis cinerea, Epicoccum nigrum, and Epicoccum sp. These differences suggest that the different products used in grapevine production have impacts in fungal endophytic composition. Further investigation of the identified fungi with respect to their antagonistic characteristics and potential use in plant protection to ensure food safety is now in course.
Resumo:
The production of activated carbons (ACs) involves two main steps: the carbonization of the carbonaceous of raw materials at temperatures below 1073 K in the absence of oxygen and the activation had realized at the temperature up to 1173 but the most useful temperature at 1073 K. In our study we used the most common industrial and consumer solid waste, namely PET, alone or blended with other synthetic polymer PAN. By mixing the two polymers in different ratios, an improvement of the yield of the AC production was found and some textural properties were enhanced by comparison with the AC prepared using each polymer separately. When all the samples were exposed through the carbonization process with a pyrolysis the mixture of PAN-PET (1:1w/w) yield around 31.9%, between that obtained with PET (16.9%) or PAN (42.6%) separately. The combine activation, with CO2 at 1073 K, allow ACs with a lower burn-off degree isothermally, when compared with those attained with PET or PAN alone, but with similarly chemicals or textural properties. The resultant ACs are microporous in their nature, as the activation time increase, the PET-PAN mixture AC are characterized by a better developed porous structure, when associated with the AC prepared from PAN. The AC prepared from PET-PAN mixture are characterized by basic surface characteristics, with a pHpzc around 10.5, which is an important characteristic for future applications on acidic pollutants removals from liquid or gaseous phase. In this study we had used the FTIR methods to determine the main functional groups in the surface of the activated carbons. The adsorbents prepared from PAN fibres presents an IR spectrum with similar characteristics to those obtained with PET wastes, but with fewer peaks and bands with less intensity, in particular for the PAN-8240 sample. This can be reflected by the stretching and deformation modes of NH bond in the range 3100 – 3300 cm-1 and 1520 – 1650 cm-1, respectively. Also, stretching mode associated to C–N, C=N, can contributed to the profile of IR spectrum around 1170 cm-1, 1585 – 1770 cm-1. And the TGA methods was used to study the loses of the precursors mass according to the excessive of the temperature. The results showed that, there were different decreasing of the mass of each precursors. PAN degradation started at almost 573 K and at 1073 K, PAN preserve more than 40% of the initial mass. PET degradation started at 650 K, but at 1073 K, it has lost 80% of the initial mass. However, the mixture of PET-PAN (1:1w/w) showed a thermogravimetric profile between the two polymers tested individually, with a final mass slightly less than 30%. From a chemical point of view, the carbonisation of PET mainly occurs in one step between 650 and 775 K.
Resumo:
Pine wilt disease (PWD) results from the interaction of three elements: the pathogenic nematode, Bursaphelenchus xylophilus; the insect-vector, Monochamus sp.; and the host tree, mostly Pinus species. Bacteria isolated from B. xylophilus may be a fourth element in this complex disease. However, the precise role of bacteria in this interaction is unclear as both plant-beneficial and as plant-pathogenic bacteria may be associated with PWD. Using whole genome sequencing and phenotypic characterization, we were able to investigate in more detail the genetic repertoire of Serratia marcescens PWN146, a bacterium associated with B. xylophilus. We show clear evidence that S. marcescens PWN146 is able to withstand and colonize the plant environment, without having any deleterious effects towards a susceptible host (Pinus thunbergii), B. xylophilus nor to the nematode model C. elegans. This bacterium is able to tolerate growth in presence of xenobiotic/organic compounds, and use phenylacetic acid as carbon source. Furthermore, we present a detailed list of S. marcescens PWN146 potentials to interfere with plant metabolism via hormonal pathways and/or nutritional acquisition, and to be competitive against other bacteria and/or fungi in terms of resource acquisition or production of antimicrobial compounds. Further investigation is required to understand the role of bacteria in PWD. We have now reinforced the theory that B. xylophilus-associated bacteria may have a plant origin.
Resumo:
Sunflower cropped area in Brazil has been showing potential possibilities to be increased in a short period of time for biofuel production. Planning the activities is one of the requirements for the success of future cropped area expansion. This requires a previous survey that identifies future trends in the transformation and rearrangement of the sunflower agro-industry sector and also identifies technological needs that may affect this process. With the objectives of identify future trends and technological needs, a value production chain was built and a questionary was applied to agents of all the sectors participating at the V National Brazilian Symposium of Sunflower and at the XVII Sunflower National Research Meeting Network. The results pointed out a strong tendency for area expansion in the next two to five years (75%); being as a secondary follow-up crop (83%) specially after soybean and top be used for biofuel (77%). The main research needs were linked with disease control, crop zoning and varietal improvement for disease resistance and high oleic oil content. Also considering the vision of and concerns regarding the future expansion and transformation of the sunflower productive complex, it is believed that the expansion is a consolidated trend, requiring a strategic sector planning associated with an economic and technological police for its success within the Brazilian agribusiness.
Resumo:
Temperate fruit production is an important economic activity in Southern Brazil. In this region, persimmon Diospyros kaki L. (Ebenaceae) is grown in small farms for domestic consumption and internal market. Mealybugs are o*en found in at least 50% of the orchards causing damages on fruits due to the honeydew secretion. However, there is a lack of informatin about species composition damaging orchards in Southern Brazil. In this work, we present a survey of mealybugs associated with persimmon trees in the Serra Gaúcha Region, Rio Grande do Sul, Brazil.
Resumo:
Against a backdrop of rapidly increasing worldwide population and growing energy demand, the development of renewable energy technologies has become of primary importance in the effort to reduce greenhouse gas emissions. However, it is often technically and economically infeasible to transport discontinuous renewable electricity for long distances to the shore. Another shortcoming of non-programmable renewable power is its integration into the onshore grid without affecting the dispatching process. On the other hand, the offshore oil & gas industry is striving to reduce overall carbon footprint from onsite power generators and limiting large expenses associated to carrying electricity from remote offshore facilities. Furthermore, the increased complexity and expansion towards challenging areas of offshore hydrocarbons operations call for higher attention to safety and environmental protection issues from major accident hazards. Innovative hybrid energy systems, as Power-to-Gas (P2G), Power-to-Liquid (P2L) and Gas-to-Power (G2P) options, implemented at offshore locations, would offer the opportunity to overcome challenges of both renewable and oil & gas sectors. This study aims at the development of systematic methodologies based on proper sustainability and safety performance indicators supporting the choice of P2G, P2L and G2P hybrid energy options for offshore green projects in early design phases. An in-depth analysis of the different offshore hybrid strategies was performed. The literature reviews on existing methods proposing metrics to assess sustainability of hybrid energy systems, inherent safety of process routes in conceptual design stage and environmental protection of installations from oil and chemical accidental spills were carried out. To fill the gaps, a suite of specific decision-making methodologies was developed, based on representative multi-criteria indicators addressing technical, economic, environmental and societal aspects of alternative options. A set of five case-studies was defined, covering different offshore scenarios of concern, to provide an assessment of the effectiveness and value of the developed tools.
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In this study, the production of bioactive secondary metabolites called "allelochemicals" by algae has been investigated, specifically focusing on polyunsaturated aldehydes (PUAs). PUAs are known to have adverse effects on planktonic grazers and on phytoplankton; however, their effect on benthic communities has been poorly studied. Macroalgae are ecosystem engineers that play an important role in the structure of the habitat and associated communities, presenting a great variability in their morphology and structural complexity, which is a primary factor in the structuring of associated communities. In recent decades, it has been seen how the introduction of invasive species can modify the benthic habitat structure, causing cascading effects on the trophic chain. The thesis includes several field and laboratory studies. Field studies examined aldehyde production by native and invasive macroalgal species (Sargassum muticum, in the Adriatic Sea, and Rugulopterix okamurae in the Strait of Gibraltar), their structural complexity, together with their associated phyto and meiobenthos. Two laboratory studies were conducted. The first one, based on microcosms experiments, evaluated the effect of PUA (produced by the diatom Skeletonema marinoi, or as decadienal analytical standard) on meiofauna. The second one evaluated the inhibitory effect of dilkamural, an allelopathic compound isolated from R. okamurae, on unicellular phototrophs. Our results showed that PUAs produced by macroalgae were species-specific and had a significant impact on the benthic community. The morphology of macroalgae was an important factor in shaping associated communities, particularly for microphytobenthos. Invasive species, such as S. muticum and R. okamurae, could reduce the biodiversity of native benthic communities and simplify the habitat. Dilkamural was hypothesized to be an allelochemical defense, and laboratory toxicity tests confirmed this hypothesis. Overall, this thesis sheds light on the importance of allelochemicals and macroalgal structural complexity in the benthic environment and highlights the potential impact of invasive species.
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The rate at which petroleum based plastics are being produced, used and thrown away is increasing every year because of an increase in the global population. Polyhydroxyalkanoates can represent a valid alternative to petroleum based plastics. They are biodegradable polymers that can be produced by some microorganisms as intracellular reserves. The actual problem is represented by the production cost of these bioplastics, which is still not competitive if compared to the one of petroleum based plastics. Mixed microbial cultures can be fed with substrates obtained from the acidogenic fermentation of carbon rich wastes, such as cheese whey, municipal effluents and various kinds of food wastes, that have a low or sometimes even inexisting cost and in this way wastes can be valorized instead of being discharged. The process consists of three phases: acidogenic fermentation in which the substrate is obtained, culture selection in which a PHA-storing culture is selected and enriched eliminating organisms that do not show this property and accumulation, in which the culture is fed until reaching the maximum storage capacity. In this work the possibility to make the process cheaper was explored trying to couple the selection and accumulation steps and a halotolerant culture collected from seawater was used and fed with an artificially salted synthetic substrated made of an aqueous solution containing a mixture of volatile fatty acids in order to explore also if its performance can allow to use it to treat substrates derived from saline effluents, as these streams cannot be treated properly by bacterias found in activated sludge plants due to inhibition caused by high salt concentrations. Generating and selling the produced PHAs obtained from these bacterias it could be possible to lower, nullify or even overcome the costs associated to the new section of a treating plant dedicated to saline effluents.
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Phospholipases A2 (PLA2) are key enzymes for production of lipid mediators. We previously demonstrated that a snake venom sPLA2 named MT-III leads to prostaglandin (PG)E2 biosynthesis in macrophages by inducing the expression of cyclooxygenase-2 (COX-2). Herein, we explored the molecular mechanisms and signaling pathways leading to these MT-III-induced effects. Results demonstrated that MT-III induced activation of the transcription factor NF-κB in isolated macrophages. By using NF-κB selective inhibitors, the involvement of this factor in MT-III-induced COX-2 expression and PGE2 production was demonstrated. Moreover, MT-III-induced COX-2 protein expression and PGE2 release were attenuated by pretreatment of macrophages with SB202190, and Ly294002, and H-7-dihydro compounds, indicating the involvement of p38MAPK, PI3K, and PKC pathways, respectively. Consistent with this, MT-III triggered early phosphorylation of p38MAPK, PI3K, and PKC. Furthermore, SB202190, H-7-dihydro, but not Ly294002 treatment, abrogated activation of NF-κB induced by MT-III. Altogether, these results show for the first time that the induction of COX-2 protein expression and PGE2 release, which occur via NF-κB activation induced by the sPLA2-MT-III in macrophages, are modulated by p38MAPK and PKC, but not by PI3K signaling proteins.
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Hippocampal sclerosis (HS) is considered the most frequent neuropathological finding in patients with mesial temporal lobe epilepsy (MTLE). Hippocampal specimens of pharmacoresistant MTLE patients that underwent epilepsy surgery for seizure control reveal the characteristic pattern of segmental neuronal cell loss and concomitant astrogliosis. However, classification issues of hippocampal lesion patterns have been a matter of intense debate. International consensus classification has only recently provided significant progress for comparisons of neurosurgical and clinic-pathological series between different centers. The respective four-tiered classification system of the International League Against Epilepsy subdivides HS into three types and includes a term of gliosis only, no-HS. Future studies will be necessary to investigate whether each of these subtypes of HS may be related to different etiological factors or with postoperative memory and seizure outcome. Molecular studies have provided potential deeper insights into the pathogenesis of HS and MTLE on the basis of epilepsy-surgical hippocampal specimens and corresponding animal models. These include channelopathies, activation of NMDA receptors, and other conditions related to Ca(2+) influx into neurons, the imbalance of Ca(2+)-binding proteins, acquired channelopathies that increase neuronal excitability, paraneoplastic and non-paraneoplastic inflammatory events, and epigenetic regulation promoting or facilitating hippocampal epileptogenesis. Genetic predisposition for HS is clearly suggested by the high incidence of family history in patients with HS, and by familial MTLE with HS. So far, it is clear that HS is multifactorial and there is no individual pathogenic factor either necessary or sufficient to generate this intriguing histopathological condition. The obvious variety of pathogenetic combinations underlying HS may explain the multitude of clinical presentations, different responses to clinical and surgical treatment. We believe that the stratification of neuropathological patterns can help to characterize specific clinic-pathological entities and predict the postsurgical seizure control in an improved fashion.
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To estimate the impact of aging and diabetes on insulin sensitivity, beta-cell function, adipocytokines, and incretin production. Hyperglycemic clamps, arginine tests and meal tolerance tests were performed in 50 non-obese subjects to measure insulin sensitivity (IS) and insulin secretion as well as plasma levels of glucagon, GLP-1 and GIP. Patients with diabetes and healthy control subjects were divided into the following groups: middle-aged type 2 diabetes (MA-DM), aged Type 2 diabetes (A-DM) and middle-aged or aged subjects with normal glucose tolerance (MA-NGT or A-NGT). IS, as determined by the homeostasis model assessment, glucose infusion rate, and oral glucose insulin sensitivity, was reduced in the aged and DM groups compared with MA-NGT, but it was similar in the MA-DM and A-DM groups. Insulinogenic index, first and second phase insulin secretion and the disposition indices, but not insulin response to arginine, were reduced in the aged and DM groups. Postprandial glucagon production was higher in MA-DM compared to MA-NGT. Whereas the GLP-1 production was reduced in A-DM, no differences between groups were observed in GIP production. In non-obese subjects, diabetes and aging impair insulin sensitivity. Insulin production is reduced by aging, and diabetes exacerbates this condition. Aging associated defects superimposed diabetic physiopathology, particularly regarding GLP-1 production. On the other hand, the glucose-independent secretion of insulin was preserved. Knowledge of the complex relationship between aging and diabetes could support the development of physiopathological and pharmacological based therapies.