Kinetic Analysis of Gill (Na+,K+)-ATPase Activity in Selected Ontogenetic Stages of the Amazon River Shrimp, (Decapoda, Palaemonidae): Interactions at ATP- and Cation-Binding Sites

We investigated modulation by ATP, Mg2+, Na+, K+ and NH4 (+) and inhibition by ouabain of (Na+,K+)-ATPase activity in microsomal homogenates of whole zoeae I and decapodid III (formerly zoea IX) and whole-body and gill homogenates of juvenile and adult Amazon River shrimps, . (Na+,K+)-ATPase-specific activity was increased twofold in decapodid III compared to zoea I, juveniles and adults, suggesting an important role in this ontogenetic stage. The apparent affinity for ATP ( (M) = 0.09 +/- A 0.01 mmol L-1) of the decapodid III (Na+,K+)-ATPase, about twofold greater than the other stages, further highlights this relevance. Modulation of (Na+,K+)-ATPase activity by K+ also revealed a threefold greater affinity for K+ ( (0.5) = 0.91 +/- A 0.04 mmol L-1) in decapodid III than in other stages; NH4 (+) had no modulatory effect. The affinity for Na+ ( (0.5) = 13.2 +/- A 0.6 mmol L-1) of zoea I (Na+,K+)-ATPase was fourfold less than other stages. Modulation by Na+, Mg2+ and NH4 (+) obeyed cooperative kinetics, while K+ modulation exhibited Michaelis-Menten behavior. Rates of maximal Mg2+ stimulation of ouabain-insensitive ATPase activity differed in each ontogenetic stage, suggesting that Mg2+-stimulated ATPases other than (Na+,K+)-ATPase are present. Ouabain inhibition suggests that, among the various ATPase activities present in the different stages, Na+-ATPase may be involved in the ontogeny of osmoregulation in larval The NH4 (+)-stimulated, ouabain-insensitive ATPase activity seen in zoea I and decapodid III may reflect a stage-specific means of ammonia excretion since functional gills are absent in the early larval stages.

The human organic cation transporter OCT1 mediates high affinity uptake of the anticancer drug daunorubicin

Les anthracyclines tels que la doxorubicin et la daunorubicin sont une famille de médicaments anticancéreux hydrophiles qui doivent être transportés dans les cellules afin d’exercer leur action par intercalation à l’ADN dans le noyau cellulaire. Ceci mène à la perturbation du métabolisme de l’ADN et entraine la mort cellulaire. Les anthracyclines sont utilisés pour le traitement d’une variété de cancers incluant la leucémie, les lymphomes, le cancer du sein, le cancer des poumons et le cancer des ovaires. Étant donné que le transport actif des anthracyclines dans les cellules a partiellement été démontré, le transporteur spécifique impliqué dans ce processus n’est pas encore connu. En utilisant un modèle de cancer des ovaires, la lignée cellulaire TOV2223G, nous avons démontré que des substrats spécifiques au transporteur de cations organiques 1 (OCT1), notamment la ergothionéine, la thiamine et la phenformin, ont partiellement inhibé l’absorption de la daunorubicin en différence de la carnitine qui est un substrat de haute affinité des transporteurs CT2 et OCTN2. Ces résultats suggèrent que les transporteurs organiques spécifiques au transport de la carnitine ne sont pas impliqués dans le transport des anthracyclines. Ainsi, nos résultats ont démontré que l’absorption de la daunorubicin est orchestrée par le transporteur OCT1 dans les cellules TOV2223G (Km ~ 5 μM) et des concentrations micromolaires de choline ont complètement abolies l’absorption de la drogue. De plus, un ARN sh dirigé contre OCT1 a réprimé son expression protéique, ce qui a été confirmé par la technique d’immuno-buvardage en utilisant un anti-OCT1 anticorps. Les cellules déficientes en OCT1 n’ont pas été capables d’absorber la daunorubicin et ont été plus résistantes à l’action de la drogue par rapport aux cellules contrôle. La transfection des cellules HEK293T avec un plasmide construit de façon à faire exprimer OCT1 comme protéine de fusion avec la protéine fluorescente EYFP a montré que celle-ci est localisée dans la membrane plasmique. Les cellules transfectées ont été capables d’absorber cinq fois plus de daunorubicin comparé aux cellules contrôles. Cette étude est, selon nous, la première à démontrer que OCT1 est un transporteur de haute affinité des anthracyclines. Ainsi, nous avons émis l’hypothèse que des défauts de OCT1 peuvent contribuer à l’efficacité de la réponse des cellules cancéreuses à la chimiothérapie avec les anthracyclines.

Optical Waveguides and Structures for Short Haul Optical Communication Channels within Printed Circuit Boards

Optical waveguides have shown promising results for use within printed circuit boards. These optical waveguides have higher bandwidth than traditional copper transmission systems and are immune to electromagnetic interference. Design parameters for these optical waveguides are needed to ensure an optimal link budget. Modeling and simulation methods are used to determine the optimal design parameters needed in designing the waveguides. As a result, optical structures necessary for incorporating optical waveguides into printed circuit boards are designed and optimized. Embedded siloxane polymer waveguides are investigated for their use in optical printed circuit boards. This material was chosen because it has low absorption, high temperature stability, and can be deposited using common processing techniques. Two sizes of waveguides are investigated, 50 $unit{mu m}$ multimode and 4 - 9 $unit{mu m}$ single mode waveguides. A beam propagation method is developed for simulating the multimode and single mode waveguide parameters. The attenuation of simulated multimode waveguides are able to match the attenuation of fabricated waveguides with a root mean square error of 0.192 dB. Using the same process as the multimode waveguides, parameters needed to ensure a low link loss are found for single mode waveguides including maximum size, minimum cladding thickness, minimum waveguide separation, and minimum bend radius. To couple light out-of-plane to a transmitter or receiver, a structure such as a vertical interconnect assembly (VIA) is required. For multimode waveguides the optimal placement of a total internal reflection mirror can be found without prior knowledge of the waveguide length. The optimal placement is found to be either 60 µm or 150 µm away from the end of the waveguide depending on which metric a designer wants to optimize the average output power, the output power variance, or the maximum possible power loss. For single mode waveguides a volume grating coupler is designed to couple light from a silicon waveguide to a polymer single mode waveguide. A focusing grating coupler is compared to a perpendicular grating coupler that is focused by a micro-molded lens. The focusing grating coupler had an optical loss of over -14 dB, while the grating coupler with a lens had an optical loss of -6.26 dB.

WATER TRANSPORT THROUGH POROUS MEDIA AND IN FLOW CHANNELS OF PROTON EXCHANGE MEMBRANE FUEL CELL

Proton exchange membrane (PEM) fuel cell has been known as a promising power source for different applications such as automotive, residential and stationary. During the operation of a PEM fuel cell, hydrogen is oxidized in anode and oxygen is reduced in the cathode to produce the intended power. Water and heat are inevitable byproducts of these reactions. The water produced in the cathode should be properly removed from inside the cell. Otherwise, it may block the path of reactants passing through the gas channels and/or gas diffusion layer (GDL). This deteriorates the performance of the cell and eventually can cease the operation of the cell. Water transport in PEM fuel cell has been the subject of this PhD study. Water transport on the surface of the GDL, through the gas flow channels, and through GDL has been studied in details. For water transport on the surface of the GDL, droplet detachment has been measured for different GDL conditions and for anode and cathode gas flow channels. Water transport through gas flow channels has been investigated by measuring the two-phase flow pressure drop along the gas flow channels. As accumulated liquid water within gas flow channels resists the gas flow, the pressure drop increases along the flow channels. The two-phase flow pressure drop can reveal useful information about the amount of liquid water accumulated within gas flow channels. Liquid water transport though GDL has also been investigated by measuring the liquid water breakthrough pressure for the region between the capillary fingering and the stable displacement on the drainage phase diagram. The breakthrough pressure has been measured for different variables such as GDL thickness, PTFE/Nafion content within the GDL, GDL compression, the inclusion of a micro-porous layer (MPL), and different water flow rates through the GDL. Prior to all these studies, GDL microstructural properties have been studied. GDL microstructural properties such as mean pore diameter, pore diameter distribution, and pore roundness distribution have been investigated by analyzing SEM images of GDL samples.

The human organic cation transporter OCT1 mediates high affinity uptake of the anticancer drug daunorubicin

Les anthracyclines tels que la doxorubicin et la daunorubicin sont une famille de médicaments anticancéreux hydrophiles qui doivent être transportés dans les cellules afin d’exercer leur action par intercalation à l’ADN dans le noyau cellulaire. Ceci mène à la perturbation du métabolisme de l’ADN et entraine la mort cellulaire. Les anthracyclines sont utilisés pour le traitement d’une variété de cancers incluant la leucémie, les lymphomes, le cancer du sein, le cancer des poumons et le cancer des ovaires. Étant donné que le transport actif des anthracyclines dans les cellules a partiellement été démontré, le transporteur spécifique impliqué dans ce processus n’est pas encore connu. En utilisant un modèle de cancer des ovaires, la lignée cellulaire TOV2223G, nous avons démontré que des substrats spécifiques au transporteur de cations organiques 1 (OCT1), notamment la ergothionéine, la thiamine et la phenformin, ont partiellement inhibé l’absorption de la daunorubicin en différence de la carnitine qui est un substrat de haute affinité des transporteurs CT2 et OCTN2. Ces résultats suggèrent que les transporteurs organiques spécifiques au transport de la carnitine ne sont pas impliqués dans le transport des anthracyclines. Ainsi, nos résultats ont démontré que l’absorption de la daunorubicin est orchestrée par le transporteur OCT1 dans les cellules TOV2223G (Km ~ 5 μM) et des concentrations micromolaires de choline ont complètement abolies l’absorption de la drogue. De plus, un ARN sh dirigé contre OCT1 a réprimé son expression protéique, ce qui a été confirmé par la technique d’immuno-buvardage en utilisant un anti-OCT1 anticorps. Les cellules déficientes en OCT1 n’ont pas été capables d’absorber la daunorubicin et ont été plus résistantes à l’action de la drogue par rapport aux cellules contrôle. La transfection des cellules HEK293T avec un plasmide construit de façon à faire exprimer OCT1 comme protéine de fusion avec la protéine fluorescente EYFP a montré que celle-ci est localisée dans la membrane plasmique. Les cellules transfectées ont été capables d’absorber cinq fois plus de daunorubicin comparé aux cellules contrôles. Cette étude est, selon nous, la première à démontrer que OCT1 est un transporteur de haute affinité des anthracyclines. Ainsi, nous avons émis l’hypothèse que des défauts de OCT1 peuvent contribuer à l’efficacité de la réponse des cellules cancéreuses à la chimiothérapie avec les anthracyclines.

The user participation on facebook pages of the three Portuguese tv news channels – RTP3, SIC NOTÍCIAS, AND TVI24

In recent decades, the national and international media contexts, in particular television media, significantly changed. The role that social networks, in particular Facebook, have taken as a content diffusion platform is unquestionable. Nowadays, traditional media (radio, newspaper, television) use the Web’s potential to distribute news content (Canelas, 2011). Currently, all TV news channels in Portugal have a website or a page on social networks. TV stations have increased communication channels with the public on digital platforms and study strategies that promote the participation and interaction with the news content (Cazajeira, 2015). The TV / Internet convergence will not only reach the content, but also the consumer, who becomes an interactive and participative audience. This reality imposes on journalism a continuous and updated news production system, dependent on a user being permanently connected to the Internet (Cazajeira, 2015). In fact, a report launched by an autonomous institution that has the function of supervising and regulating the media Portugal (ERC, 2015), confirms the relevance that social media has assumed in the publication and consumption of news. Social networks are recognised as one of the most important means for news media consultation, right after television, and the practice of sharing news is very common among consumers of online news in Portugal. Furthermore, when compared to other countries analysed by Reuters Institute (Newman, Levy, & Nielsen, 2015), Portuguese consumers are those who make the most comments to online news, preferring social networks to news sites. Considering the importance of new online platforms for journalism, this study aims to present a quantitative analysis of user participation on the Facebook pages of the three Portuguese TV news channels, specifically RTP3, SIC Notícias and TVI24, between 8 and 14 February 2016. To track this participation, the following parameters were used: the "like" button as a way to study the demonstration of publication interest; "sharing" of a particular element, be it a photo, a video or a text, on the user Timeline, the Timeline of a friend or by private message. This monitoring is important to understand the dissemination of news content; and the comments area. The number of comments will help understand the dynamics and the discussion that the publication has on the public. The results of 1063 posts indicate that of the analysed parameters - "Like", "Comment", and "Share" – the one with the greatest power of participation among the users of the pages of the three Portuguese TV news channels is the "Like" system, followed by "Share" and then "Comment". The theme that generates the most user participation with "Likes" and “Comments” parameters are "Science and Technology", “Education” and “Humorous/Satirical/Unusual”. Finally, the publications available end of the night (10pm-1am) has better participation rates.

The presence of three Portuguese TV news channels (RTP3, SIC NOTÍCIAS, and TVI24) on facebook

The growing popularity of social networks, and their impact on the daily lives of consumers, contributed to news organizations marking their presence on different online platforms. In the case of Facebook, a social network that began as a personal space, it has gradually transformed into a content-sharing space (Oeldorf-Hirsch & Sundar, 2015). Nowadays, Facebook is the second most viewed website in Portugal (Alexa, 2016), and therefore, it has become crucial for Portuguese news agencies to be present on this social network. Although television continues to be the main information source in Portugal, social networks, and specifically Facebook, are increasingly important in news consumption by users (ERC, 2015). This new way of news dissemination, as well as the proliferation that these contents reach in social networks, led to news agencies exploiting these new channels, both to attract new audiences, and to redirect users to their own websites (Castillo, El-Haddad, Pfeffer, & Stempeck, 2014). Thus, it is important to understand how, and what kind of content these agencies put on their Facebook channels, as well as the strategies they use to share these same contents. This study aims to understand how the main news channels of Portuguese TV (RTP3, SIC Notícias, and TVI24) manage and use the social network Facebook to share news contents. To this end, the authors collected quantitative data of all posts placed on Facebook between February 8 and February 14 2016. Approximately 1063 posts were collected and analysed from the three Facebook pages. The results indicate that two of the three channels extensively used their Facebook pages to share and target content to their official websites. Regarding the news sources and type of media used, the three Portuguese TV news channels use similar strategies. However, in what concerns the main themes and quantity of messages per day, as well as the level of redundancy of information, the three channels manage their pages differently.

The presence of three portuguese TV news channels (RTP3, SIC Notícias, and TVI24) on Facebook

The growing popularity of social networks, and their impact on the daily lives of consumers, contributed to news organizations marking their presence on different online platforms. In the case of Facebook, a social network that began as a personal space, it has gradually transformed into a content-sharing space (Oeldorf-Hirsch & Sundar, 2015). Nowadays, Facebook is the second most viewed website in Portugal (Alexa, 2016), and therefore, it has become crucial for Portuguese news agencies to be present on this social network. Although television continues to be the main information source in Portugal, social networks, and specifically Facebook, are increasingly important in news consumption by users (ERC, 2015). This new way of news dissemination, as well as the proliferation that these contents reach in social networks, led to news agencies exploiting these new channels, both to attract new audiences, and to redirect users to their own websites (Castillo, El-Haddad, Pfeffer, & Stempeck, 2014). Thus, it is important to understand how, and what kind of content these agencies put on their Facebook channels, as well as the strategies they use to share these same contents. This study aims to understand how the main news channels of Portuguese TV (RTP3, SIC Notícias, and TVI24) manage and use the social network Facebook to share news contents. To this end, the authors collected quantitative data of all posts placed on Facebook between February 8 and February 14 2016. Approximately 1063 posts were collected and analysed from the three Facebook pages. The results indicate that two of the three channels extensively used their Facebook pages to share and target content to their official websites. Regarding the news sources and type of media used, the three Portuguese TV news channels use similar strategies. However, in what concerns the main thematic and quantity of messages per day, as well as the level of redundancy of information, the three channels operate their pages differently.

The presence of three Portuguese TV news channels (RTP3, SIC NOTÍCIAS, and TVI24) on facebook

In recent decades, the national and international media contexts, in particular television, media significantly changed. Although they were not created for the purpose, the role that social networks, in particular Facebook, have taken as a content diffusion platform is unquestionable. Nowadays, traditional media (radio, newspaper, television) use the Web’s potential to distribute news content (Canelas, 2011). Currently, all TV news channels in Portugal have a website or a page on social networks. TV stations have increased communication channels with the public on digital platforms and study strategies that promote the participation and interaction with the news content (Cazajeira, 2015). The TV / Internet convergence will not only reach the content, but also the consumer, who becomes an interactive and participative audience. This reality imposes on journalism a continuous and updated news production system, dependent on a user being permanently connected to the Internet (Cazajeira, 2015). In fact, an ERC report (2015) confirms the relevance that social media has assumed in the publication and consumption of news. Social networks are recognised as one of the most important means for news media consultation, right after television, and the practice of sharing news is very common among consumers of online news in Portugal. Furthermore, when compared to other countries analysed by Reuters Institute (Newman, Levy, & Nielsen, 2015), Portuguese consumers are those who make the most comments to online news, preferring social networks to news sites. Considering the importance of new online platforms for journalism, this study aims to present a quantitative analysis of user participation on the Facebook pages of the three Portuguese TV news channels, specifically RTP3, SIC Notícias and TVI24, between 8 and 14 February 2016. To track this participation, the following parameters were used: the "like" button as a way to study the demonstration of publication interest; "sharing" of a particular element, be it a photo, a video or a text, on their own Timeline, the Timeline of a friend or by private message. This monitoring is important to understand the dissemination of news content; and the comments area. The number of comments will help understand the dynamics and the discussion that the publication has on the public. The results of 1063 posts indicate that of the analysed parameters - "Like", "Comment", and "Share" – the one with the greatest power of participation among the users of the pages of the three Portuguese TV news channels is the "Like" system, followed by "Share" and then "Comment". Analysing the thematic that create the most user participation with "Likes" and “Comments” parameters are "Science and Technology". “Education” and “Humorous/Satirical/Unusual Finally, the publications available end of the night (10pm-1am) has better participation rates.

Classi cation of new electricity customers based on surveys and smart metering data

This paper proposes a process for the classifi cation of new residential electricity customers. The current state of the art is extended by using a combination of smart metering and survey data and by using model-based feature selection for the classifi cation task. Firstly, the normalized representative consumption profi les of the population are derived through the clustering of data from households. Secondly, new customers are classifi ed using survey data and a limited amount of smart metering data. Thirdly, regression analysis and model-based feature selection results explain the importance of the variables and which are the drivers of diff erent consumption profi les, enabling the extraction of appropriate models. The results of a case study show that the use of survey data signi ficantly increases accuracy of the classifi cation task (up to 20%). Considering four consumption groups, more than half of the customers are correctly classifi ed with only one week of metering data, with more weeks the accuracy is signifi cantly improved. The use of model-based feature selection resulted in the use of a signifi cantly lower number of features allowing an easy interpretation of the derived models.

Exploiting molecular simulations as an atomic-level microscope to elucidate clinically relevant cation chloride cotransporters

The cation chloride cotransporters (CCCs) represent a vital family of ion transporters, with several members implicated in significant neurological disorders. Specifically, conditions such as cerebrospinal fluid accumulation, epilepsy, Down’s syndrome, Asperger’s syndrome, and certain cancers have been attributed to various CCCs. This thesis delves into these pharmacological targets using advanced computational methodologies. I primarily employed GPU-accelerated all-atom molecular dynamics simulations, deep learning-based collective variables, enhanced sampling methods, and custom Python scripts for comprehensive simulation analyses. Our research predominantly centered on KCC1 and NKCC1 transporters. For KCC1, I examined its equilibrium dynamics in the presence/absence of an inhibitor and assessed the functional implications of different ion loading states. In contrast, our work on NKCC1 revealed its unique alternating access mechanism, termed the rocking-bundle mechanism. I identified a previously unobserved occluded state and demonstrated the transporter's potential for water permeability under specific conditions. Furthermore, I confirmed the actual water flow through its permeable states. In essence, this thesis leverages cutting-edge computational techniques to deepen our understanding of the CCCs, a family of ion transporters with profound clinical significance.

Multivariate analysis to discriminate top quark pair production channels at LHC

Il quark top è una delle particelle fondamentali del Modello Standard, ed è osservato a LHC nelle collisioni a più elevata energia. In particolare, la coppia top-antitop (tt̄) è prodotta tramite interazione forte da eventi gluone-gluone (gg) oppure collisioni di quark e antiquark (qq̄). I diversi meccanismi di produzione portano ad avere coppie con proprietà diverse: un esempio è lo stato di spin di tt̄, che vicino alla soglia di produzione è maggiormente correlato nel caso di un evento gg. Uno studio che voglia misurare l’entità di tali correlazioni risulta quindi essere significativamente facilitato da un metodo di discriminazione delle coppie risultanti sulla base del loro canale di produzione. Il lavoro qui presentato ha quindi lo scopo di ottenere uno strumento per effettuare tale differenziazione, attraverso l’uso di tecniche di analisi multivariata. Tali metodi sono spesso applicati per separare un segnale da un fondo che ostacola l’analisi, in questo caso rispettivamente gli eventi gg e qq̄. Si dice che si ha a che fare con un problema di classificazione. Si è quindi studiata la prestazione di diversi algoritmi di analisi, prendendo in esame le distribuzioni di numerose variabili associate al processo di produzione di coppie tt̄. Si è poi selezionato il migliore in base all’efficienza di riconoscimento degli eventi di segnale e alla reiezione degli eventi di fondo. Per questo elaborato l’algoritmo più performante è il Boosted Decision Trees, che permette di ottenere da un campione con purezza iniziale 0.81 una purezza finale di 0.92, al costo di un’efficienza ridotta a 0.74.

Taurine Supplementation Increases K(atp) Channel Protein Content, Improving Ca2+ Handling And Insulin Secretion In Islets From Malnourished Mice Fed On A High-fat Diet.

Pancreatic β-cells are highly sensitive to suboptimal or excess nutrients, as occurs in protein-malnutrition and obesity. Taurine (Tau) improves insulin secretion in response to nutrients and depolarizing agents. Here, we assessed the expression and function of Cav and KATP channels in islets from malnourished mice fed on a high-fat diet (HFD) and supplemented with Tau. Weaned mice received a normal (C) or a low-protein diet (R) for 6 weeks. Half of each group were fed a HFD for 8 weeks without (CH, RH) or with 5% Tau since weaning (CHT, RHT). Isolated islets from R mice showed lower insulin release with glucose and depolarizing stimuli. In CH islets, insulin secretion was increased and this was associated with enhanced KATP inhibition and Cav activity. RH islets secreted less insulin at high K(+) concentration and showed enhanced KATP activity. Tau supplementation normalized K(+)-induced secretion and enhanced glucose-induced Ca(2+) influx in RHT islets. R islets presented lower Ca(2+) influx in response to tolbutamide, and higher protein content and activity of the Kir6.2 subunit of the KATP. Tau increased the protein content of the α1.2 subunit of the Cav channels and the SNARE proteins SNAP-25 and Synt-1 in CHT islets, whereas in RHT, Kir6.2 and Synt-1 proteins were increased. In conclusion, impaired islet function in R islets is related to higher content and activity of the KATP channels. Tau treatment enhanced RHT islet secretory capacity by improving the protein expression and inhibition of the KATP channels and enhancing Synt-1 islet content.