Search results for: Emilio Esposito

Authors: Chang-Hui Shen, Michelle Esposito, Andrew J. Shen, Michael Adejokun, Diana Laterman

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The packaging of DNA into nucleosomes is critical to genomic compaction, yet it can leave gene promoters inaccessible to activator proteins or transcription machinery and thus prevents transcriptional initiation. Both chromatin remodelers and histone acetylases (HATs) are the two main transcription co-activators that can reconfigure chromatin structure for transcriptional activation. Ino80p is the core component of the INO80 remodeling complex. Recently, it was shown that Ino80p dissociates from the yeast INO1 promoter after induction. However, when certain HATs were deleted or mutated, Ino80p accumulated at the promoters during gene activation. This suggests a link between HATs’ presence and Ino80p’s dissociation. However, it has yet to be demonstrated that Ino80p can be acetylated. To determine if Ino80p can be acetylated, wild-type Saccharomyces cerevisiae cells carrying Ino80p engineered with a double FLAG tag (MATa INO80-FLAG his3∆200 leu2∆0 met15∆0 trp1∆63 ura3∆0) were grown to mid log phase, as were non-tagged wild type (WT) (MATa his3∆200 leu2∆0 met15∆0 trp1∆63 ura3∆0) and ino80∆ (MATa ino80∆::TRP1 his3∆200 leu2∆0 met15∆0 trp1∆63 ura3∆0) cells as controls. Cells were harvested, and the cell lysates were subjected to immunoprecipitation (IP) with α-FLAG resin to isolate Ino80p. These eluted IP samples were subjected to SDS-PAGE and Western blot analysis. Subsequently, the blots were probed with the α-FLAG and α-acetyl lysine antibodies, respectively. For the blot probed with α-FLAG, one prominent band was shown in the INO80-FLAG cells, but no band was detected in the IP samples from the WT and ino80∆ cells. For the blot probed with the α-acetyl lysine antibody, we detected acetylated Ino80p in the INO80-FLAG strain while no bands were observed in the control strains. As such, our results showed that Ino80p can be acetylated. This acetylation can explain the co-activator’s recruitment patterns observed in current gene activation models. In yeast INO1, it has been shown that Ino80p is recruited to the promoter during repression, and then dissociates from the promoter once de-repression begins. Histone acetylases, on the other hand, have the opposite pattern of recruitment, as they have an increased presence at the promoter as INO1 de-repression commences. This Ino80p recruitment pattern significantly changes when HAT mutant strains are studied. It was observed that instead of dissociating, Ino80p accumulates at the promoter in the absence of functional HATs, such as Gcn5p or Esa1p, under de-repressing processes. As such, Ino80p acetylation may be required for its proper dissociation from the promoters. The remodelers’ dissociation mechanism may also have a wide range of implications with respect to transcriptional initiation, elongation, or even repression as it allows for increased spatial access to the promoter for the various transcription factors and regulators that need to bind in that region. Our findings here suggest a previously uncharacterized interaction between Ino80p and other co-activators recruited to promoters. As such, further analysis of Ino80p acetylation not only will provide insight into the role of epigenetic modifications in transcriptional activation, but also gives insight into the interactions occurring between co-activators at gene promoters during gene regulation.

Keywords: acetylation, chromatin remodeler, epigenetic modification, Ino80p

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Authors: Marcio Emilio Dos Santos, Kelly C. F. Dos Santos

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Caregivers of patients diagnosed with ASD are subjected to high stress situations due to the complexity and multiple levels of daily activities that require the organization of events, behaviors and socioemotional situations, such as immediate decision making and in public spaces. The cognitive and emotional requirement needed to fulfill this caregiving role exceeds the regular cultural process that adults receive in their process of preparation for conjugal and parental life. Therefore, in many cases, caregivers present a high level of overload, poor capacity to organize and mediate the development process of the child or patient about their care. Aims: Improvement in the cognitive and emotional capacities related to the caregiver function, allowing the reduction of the overload, the feeling of incompetence and the characteristic level of stress, developing a more organized conduct and decision making more oriented towards the objectives and procedural gains necessary for the integral development of the patient with diagnosis of ASD. Method: The study was performed with 20 relatives, randomly selected from a total of 140 patients attended. The family members were submitted to the Wechsler Adult Intelligence Scale III intelligence test and the Family assessment Management Measure (FaMM) questionnaire as a previous evaluation. Therapeutic activity in a small group of family members or caregivers, with weekly frequency, with a minimum workload of two hours, using the Feuerstein Instrumental Enrichment Cognitive Development Program - Feuerstein Instrumental Enrichment for ten months. Reapplication of the previous tests to verify the gains obtained. Results and Discussion: There is a change in the level of caregiver overload, improvement in the results of the Family assessment Management Measure and highlight to the increase of performance in the cognitive aspects related to problem solving, planned behavior and management of behavioral crises. These results lead to the discussion of the need to invest in the integrated care of patients and their caregivers, mainly by enabling cognitively to deal with the complexity of Autism. This goes beyond the simple therapeutic orientation about adjustments in family and school routines. The study showed that when the caregiver improves his/her capacity of management, the results of the treatment are potentiated and there is a reduction of the level of the caregiver's overload. Importantly, the study was performed for only ten months and the number of family members attended in the study (n = 20) needs to be expanded to have statistical strength.

Keywords: caregiver overload, cognitive development program ASD caregivers, feuerstein instrumental enrichment, family assessment management measure

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Authors: Einat Waizbard, Emilio Ferrer, Meghan Miller, Brianna Heath, Derek S. Andrews, Sally J. Rogers, Christine Wu Nordahl, Marjorie Solomon, David G. Amaral

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Background: Autism symptoms are comprised of social-communication deficits and restricted/repetitive behaviors (RRB). The severity of these symptoms can change during childhood, with differences between boys and girls. From the literature, it was found that young autistic girls show a stronger tendency to decrease and a weaker tendency to increase their overall autism symptom severity levels compared to young autistic boys. It is not clear, however, which symptoms are driving these sex differences across childhood. In the current study, we evaluated the trajectories of independent autism symptoms across childhood and compared the patterns of change in such symptoms between boys and girls. Method: The study included 183 children diagnosed with autism (55 girls) evaluated three times across childhood, at ages 3, 6 and 11. We analyzed 22 independent items from the Autism Diagnostic Observation Scheudule-2 (ADOS-2), the gold-standard assessment tool for autism symptoms, each item representing a specific autism symptom. First, we used latent growth curve models to estimate the trajectories for the 22 ADOS-2 items for each child in the study. Second, we extracted the factor scores representing the individual slopes for each ADOS-2 item (i.e., slope representing that child’s change in that specific item). Third, we used factor analysis to identify common patterns of change among the ADOS-2 items, separately for boys and girls, i.e., which autism symptoms tend to change together and which change independently across childhood. Results: The best-emerging patterns for both boys and girls identified four common factors: three factors representative of changes in social-communication symptoms and one factor describing changes in RRB. Boys and girls showed the same pattern of change in RRB, with four items (e.g., speech abnormalities) changing together across childhood and three items (e.g., mannerisms) changing independently of other items. For social-communication deficits in boys, three factors were identified: the first factor included six items representing initiating and engaging in social-communication (e.g., quality of social overtures, conversation), the second factor included five items describing responsive social-communication (e.g., response to name) and the third factor included three items related to different aspects of social-communication (e.g., level of language). Girls’ social-communications deficits also loaded onto three factors: the first factor included five items (e.g., unusual eye contact), the second factor included six items (e.g., quality of social response), and the third factor included four items (e.g., showing). Some items showed similar patterns of change for both sexes (e.g., responsive joint attention), while other items showed differences (e.g., shared enjoyment). Conclusions: Girls and boys had different patterns of change in autism symptom severity across childhood. For RRB, both sexes showed similar patterns. For social-communication symptoms, however, there were both similarities and differences between boys and girls in the way symptoms changed over time. The strongest patterns of change were identified for initiating and engaging in social communication for boys and responsive social communication for girls.

Keywords: autism spectrum disorder, autism symptom severity, symptom trajectories, sex differences

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Authors: Léa Caban, Lucile Soudani, Julien Berger, Armelle Nouviaire, Emilio Bastidas-Arteaga

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Improvement of the thermal performance of buildings is a high concern for the construction industry. With the increase in environmental issues, new types of construction materials are being developed. These include bio-based insulation materials. They capture carbon dioxide, can be produced locally, and have good thermal performance. However, their behavior with respect to moisture transfer is still facing some issues. With a high porosity, the mass transfer is more important in those materials than in mineral insulation ones. Therefore, they can be more sensitive to moisture disorders such as mold growth, condensation risks or decrease of the wall energy efficiency. For this reason, the initial moisture content on the construction site is a piece of crucial knowledge. Measuring moisture content in a laboratory is a mastered task. Diverse methods exist but the easiest and the reference one is gravimetric. A material is weighed dry and wet, and its moisture content is mathematically deduced. Non-destructive methods (NDT) are promising tools to determine in an easy and fast way the moisture content in a laboratory or on construction sites. However, the quality and reliability of the measures are influenced by several factors. Classical NDT portable devices usable on-site measure the capacity or the resistivity of materials. Water’s electrical properties are very different from those of construction materials, which is why the water content can be deduced from these measurements. However, most moisture meters are made to measure wooden materials, and some of them can be adapted for construction materials with calibration curves. Anyway, these devices are almost never calibrated for insulation materials. The main objective of this study is to determine the reliability of moisture meters in the measurement of biobased insulation materials. The determination of which one of the capacitive or resistive methods is the most accurate and which device gives the best result is made. Several biobased insulation materials are tested. Recycled cotton, two types of wood fibers of different densities (53 and 158 kg/m3) and a mix of linen, cotton, and hemp. It seems important to assess the behavior of a mineral material, so glass wool is also measured. An experimental campaign is performed in a laboratory. A gravimetric measurement of the materials is carried out for every level of moisture content. These levels are set using a climatic chamber and by setting the relative humidity level for a constant temperature. The mass-based moisture contents measured are considered as references values, and the results given by moisture meters are compared to them. A complete analysis of the uncertainty measurement is also done. These results are used to analyze the reliability of moisture meters depending on the materials and their water content. This makes it possible to determine whether the moisture meters are reliable, and which one is the most accurate. It will then be used for future measurements on construction sites to assess the initial hygrothermal state of insulation materials, on both new-build and renovation projects.

Keywords: capacitance method, electrical resistance method, insulation materials, moisture transfer, non-destructive testing

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Authors: Juan C. Pérez-Mesa, Emilio Galdeano-Gómez, Jerónimo De Burgos-Jiménez, José F. Bienvenido-Bárcena, José F. Jiménez-Guerrero

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Environmental cost and transport congestion on roads resulting from product distribution in Europe have to lead to the creation of various programs and studies seeking to reduce these negative impacts. In this regard, apart from other institutions, the European Commission (EC) has designed plans in recent years promoting a more sustainable transportation model in an attempt to ultimately shift traffic from the road to the sea by using intermodality to achieve a model rebalancing. This issue proves especially relevant in supply chains from peripheral areas of the continent, where the supply of certain agrifood products is high. In such cases, the most difficult challenge is managing perishable goods. This study focuses on new approaches that strengthen the modal shift, as well as the reduction of externalities. This problem is analyzed by attempting to promote intermodal system (truck and short sea shipping) for transport, taking as point of reference highly perishable products (vegetables) exported from southeast Spain, which is the leading supplier to Europe. Methodologically, this paper seeks to contribute to the literature by proposing a different and complementary approach to establish a comparison between intermodal and the “only road” alternative. For this purpose, the multicriteria decision is utilized in a p-median model (P-M) adapted to the transport of perishables and to a means of shipping selection problem, which must consider different variables: transit cost, including externalities, time, and frequency (including agile response time). This scheme avoids bias in decision-making processes. By observing the results, it can be seen that the influence of the externalities as drivers of the modal shift is reduced when transit time is introduced as a decision variable. These findings confirm that the general strategies, those of the EC, based on environmental benefits lose their capacity for implementation when they are applied to complex circumstances. In general, the different estimations reveal that, in the case of perishables, intermodality would be a secondary and viable option only for very specific destinations (for example, Hamburg and nearby locations, the area of influence of London, Paris, and the Netherlands). Based on this framework, the general outlook on this subject should be modified. Perhaps the government should promote specific business strategies based on new trends in the supply chain, not only on the reduction of externalities, and find new approaches that strengthen the modal shift. A possible option is to redefine ports, conceptualizing them as digitalized redistribution and coordination centers and not only as areas of cargo exchange.

Keywords: environmental externalities, intermodal transport, perishable food, transit time

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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

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Authors: Andrés Quiroz, Emilio Hormazabal, Ana Mutis, Fernando Ortega, Manuel Chacón-Fuentes, Leonardo Parra

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Red clover (Trifolium pratense L.) is an important forage species in temperate regions of the world. The main limitation of this species worldwide is a lack of persistence related to the high mortality of plants due to a complex of biotic and abiotic factors, determining a life span of two or three seasons. Because of the importance of red clover in Chile, a red clover breeding program was started at INIA Carillanca Research Center in 1989, with the main objective of improving the survival of plants, forage yield, and persistence. The main selection criteria for selecting new varieties have been based on agronomical parameters and biotic factors. The main biotic factor associated with red clover mortality in Chile is Hylastinus obscurus (Coleoptera: Curculionidae). Both larval and adults feed on the roots, causing weakening and subsequent death of clover plants. Pesticides have not been successful for controlling infestations of this root borer. Therefore, alternative strategies for controlling this pest are a high priority for red clover producers. Currently, the role of semiochemical in the interaction between H. obscurus and red clover plants has been widely studied for our group. Specifically, from the red clover foliage has been identified limonene is eliciting repellency from the root borer. Limonene is generated in the plant from two independent biosynthetic pathways, the mevalonic acid, and deoxyxylulose pathway. Mevalonate pathway enzymes are localized in the cytosol, whereas the deoxyxylulose phosphate pathway enzymes are found in plastids. In summary, limonene can be determinated by enzymatic bioassay using GPP as substrate and by limonene synthase expression. Therefore, the main objective of this work was to study genetic variation of limonene in material provided by INIA´s Red Clover breeding program. Protein extraction was carried out homogenizing 250 mg of leave tissue and suspended in 6 mL of extraction buffer (PEG 1500, PVP-30, 20 mM MgCl2 and antioxidants) and stirred on ice for 20 min. After centrifugation, aliquots of 2.5 mL were desalted on PD-10 columns, resulting in a final volume of 3.5 mL. Protein determination was performed according to Bradford with BSA as a standard. Monoterpene synthase assays were performed with 50 µL of protein extracts transferred into gas-tight 2 mL crimp seal vials after addition of 4 µL MgCl₂ and 41 µL assay buffer. The assay was started by adding 5 µL of a GPP solution. The mixture was incubated for 30 min at 40 °C. Biosynthesized limonene was quantified in a GC equipped with a chiral column and using synthetic R and S-limonene standards. The enzymatic the production of R and S-limonene from different Superqueli-Carillanca genotypes is shown in this work. Preliminary results showed significant differences in limonene content among the genotypes analyzed. These results constitute an important base for selecting genotypes with a high content of this repellent monoterpene towards H. obscurus.

Keywords: head space, limonene enzymatic determination, red clover, Hylastinus obscurus

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Authors: Emilio Galdeano-Gómez, Juan C. Pérez-Mesa, Laura Piedra-Muñoz, María C. García-Barranco, Jesús Hernández-Rubio

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The situation in which firms engage in simultaneous cooperation and competition with each other is a phenomenon known as co-opetition. This scenario has received increasing attention in business economics and management analyses. In the domain of supply chain networks and for small and medium-sized enterprises, SMEs, these strategies are of greater relevance given the complex environment of globalization and competition in open markets. These firms face greater challenges regarding technology and access to specific resources due to their limited capabilities and limited market presence. Consequently, alliances and collaborations with both buyers and suppliers prove to be key elements in overcoming these constraints. However, rivalry and competition are also regarded as major factors in successful internationalization processes, as they are drivers for firms to attain a greater degree of specialization and to improve efficiency, for example enabling them to allocate scarce resources optimally and providing incentives for innovation and entrepreneurship. The present work aims to contribute to the literature on SMEs’ internationalization strategies. The sample is constituted by a panel data of marketing firms from the Andalusian food sector and a multivariate regression analysis is developed, measuring variables of co-opetition and international activity. The hierarchical regression equations method has been followed, thus resulting in three estimated models: the first one excluding the variables indicative of channel type, while the latter two include the international retailer chain and wholesaler variable. The findings show that the combination of several factors leads to a complex scenario of inter-organizational relationships of cooperation and competition. In supply chain management analyses, these relationships tend to be classified as either buyer-supplier (vertical level) or supplier-supplier relationships (horizontal level). Several buyers and suppliers tend to participate in supply chain networks, and in which the form of governance (hierarchical and non-hierarchical) influences cooperation and competition strategies. For instance, due to their market power and/or their closeness to the end consumer, some buyers (e.g. large retailers in food markets) can exert an influence on the selection and interaction of several of their intermediate suppliers, thus endowing certain networks in the supply chain with greater stability. This hierarchical influence may in turn allow these suppliers to develop their capabilities (e.g. specialization) to a greater extent. On the other hand, for those suppliers that are outside these networks, this environment of hierarchy, characterized by a “hub firm” or “channel master”, may provide an incentive for developing their co-opetition relationships. These results prove that the analyzed firms have experienced considerable growth in sales to new foreign markets, mainly in Europe, dealing with large retail chains and wholesalers as main buyers. This supply industry is predominantly made up of numerous SMEs, which has implied a certain disadvantage when dealing with the buyers, as negotiations have traditionally been held on an individual basis and in the face of high competition among suppliers. Over recent years, however, cooperation among these marketing firms has become more common, for example regarding R&D, promotion, scheduling of production and sales.

Keywords: co-petition networks, international supply chain, maketing agrifood firms, SMEs strategies

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