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BRU ELT JOURNAL, 1, 1, 99-112

[EN] Most proteins cannot cross the nuclear envelope by diffusion, instead they need to be actively transported by specific nuclear transport (import or export) receptors that recognize and bind specific nucleocytoplasmic transport signals in proteins. Nuclear import receptors recognize Nuclear Localization Signals (NLSs) and transport their cargo proteins to the nucleus, while nuclear export receptors recognize Nuclear Export Signals (NESs) and transport cargos to the cytoplasm. The best-studied export receptor is CRM1, which mediates the nuclear export of cargos bearing “leucine-rich” NES motifs. CRM1-mediated nuclear export is crucial for proper protein localization and to maintain cellular homeostasis. Therefore, CRM1 malfunction can be a trigger for different human diseases, and identifying CRM1 cargo proteins might be important to better understand related pathologies. However, the full CRM1 cargo spectrum is not yet completely clear. A global proteomics analysis, proposed a long list of potential CRM1 cargoes, but many remain to be validated. In the present work, a protein from this list, c-CBL, was selected for further study, as the mechanisms and signals that might modulate its subcellular distribution are still unkown. We aimed to identify novel nucleocytoplasmic transport signals in this protein. For that purpose, in silico predictions and nuclear export assays were carried out. We report here two novel functional NESs in c-CBL, and discuss the importance of this finding in the context of c-CBL’s transport, stressing the need for further research on c-CBL’s nucleocytoplasmic transport to better understand and use c- CBL as a novel target for different therapies. [EU] Proteina gehienek ezin dute gaineztadura nuklearra difusioz zeharkatu, eta nukleoko garraio- hartzaile espezifikoen bidez igaro behar dute modu aktiboan (inportazioz edo esportazioz). Hartzaile horiek proteinen garraio nukleozitoplasmatikorako seinale espezifikoak ezagutu eta lotzen dituzte. Zehazki, inportazio- hartzaileek nukleorako kokapen-seinaleak (NLS) ezagutzen dituzte eta proteinak nukleora garraiatzen dituzte; esportazio-hartzaileek, berriz, nukleotiko esportazio-seinaleak (NESak) ezagutu eta proteinak zitoplasmara garraiatzen dituzte. Gehien ikertu den esportazio-hartzailea CRM1 da, “leuzinatan aberatsak diren NESak” dituzten proteinak zitoplasmara esportatzen dituena. CRM1 bidezko esportazioa ezinbestekoa da proteinen kokapen egokirako eta zelulako homeostasia mantentzeko. Ondorioz, CRM1-en akatsek zenbait giza gaixotasun eragin ditzakete. Horregatik, CRM1-en bidez esportatzen diren proteinak identifikatzea garrantzizkoa izan daiteke horiei loturiko gaixotasunak hobeto ulertzeko. Hala ere, CRM1-en bidez esportatzen diren proteinen espektroa (zein proteina garraiatzen diren CRM1 bidez) ez da guztiz argia oraindik. Proteomika-analisi global batek CRM1- en bidez esportatzen diren proteina potentzialen zerrenda proposatu zuen, baina horietako asko balioztatzeke daude. Lan honetan, zerrendako proteina bat, c-CBL, hautatu zen sakonago ikertzeko. Izan ere, proteina horren kokapen zelularra eraentzen duten mekanismo eta sekuentziak ezezagunak dira eta beraz, gure helburua proteina honetan garraio nukleozitoplasmatikorako sekuentzia berriak bilatzea zen. Horretarako, in silico iragarpenak egin genituen, esportazio-entseguekin batera. Lan honetan bi NES funtzional berri deskribatu dira eta aurkikuntza honek c-CBL-ren garraioan duen garrantziaz eztabaidatzen da. Azkenik, c-CBL terapia ezberdinetan itu berri gisa erabili ahal izateko, bere garraioaren inguruko ikerketa gehiagoren beharra azpimarratzen da.

Emulacija je sposobnost programske opreme ali naprave, da replicira obnašanje neke druge programske opreme ali naprave. V diplomski nalogi smo s programsko opremo emulirali retro igralno konzolo NES, ki je ena izmed najbolj znanih na svetu. Emulacija tega sistema vključuje emulacijo procesorja, vhoda, grafične enote, zvočne enote in preslikovalnikov. Emulator je dodatno nadgrajen z razhroščevalnikom, ki uporabniku omogoča vpogled v sistem. Tako je diplomska naloga zanimiva za tiste, ki si želijo preizkusiti retro igralno konzolo, kot tudi za vse, ki jih zanima, kako deluje. Emulation is the capability of a software or device to mimic some other software or device. In this thesis, we demonstrate how to emulate the NES device, which is one of the most famous retro video game consoles. The emulation of the system consists of CPU, input, PPU, APU and mapper emulation. The emulator is also supported with a debugger, which can be used to see how the emulated CPU works. This thesis is interesting to those who wish to try the games, which were originally created for an old console, and to those who want to know how this console works.

The NES (nonlinear energy sink) is a new type of nonlinear tuned mass damper that is connected to the shock-absorbing main structure through strong nonlinear stiffness and viscous damping. The vibrational energy in the main structure is transferred to the NES oscillator by means of target energy transfer. A shaking table test of a 1:4 scaled RC (Reinforced Concrete) frame structure model with a new type of NES shock absorber was conducted to study the damping effect of the NES shock absorber, especially for the influence of joint strength and deformation. The NES used in this experiment has a relatively large nonlinear stiffness and a wide vibration absorption frequency band. The variation of reinforcement strains, node failure mode, and structural natural frequency of 1 story and two-layer joints of the model frame structure with NES were studied. The test results showed that NES could effectively reduce the strains of longitudinal reinforcement and stirrup in beams and columns and delay the plastic hinge development at the bottom and the top of the column. The frame model with NES installed has failures at the beam ends and shear failures at the nodes, realizing the seismic mechanism of solid columns and weak beams. Compared with ordinary seismic structures, the NES can effectively reduce the shear stress of concrete at the joints and alleviate the shear failure of joints. The final failure of the NES shock absorbing structure was the yielding of the steel bars at the bottom of the column and the crushing of the concrete at the foot of the column, and the connection between the column foot and the backplane became loose simultaneously. The decreasing rate of the vibration frequency declined due to the NES with varied broadband absorbing capability. It can be seen that the NES shock absorber not only has a good effect on reducing the seismic response of the structure, but more importantly, the damage of the structural nodes is greatly reduced, and therefore, the seismic capacity of the structure improved.

The aim of this paper is to investigate the vibration suppression effect of a nonlinear energy sink (NES) on the wind-vortex-induced vibration of deep-water jacket pipes. Based on nonlinear dynamical theories, the coupled dynamic equation of a deep-water jacket pipe with NES was established, and the nonlinear response characteristics and motion stability of the system were studied by using the multi-scale mixed harmonic balance method and Routh–Hurwitz criterion. The paper also analyzed the influence of the nonlinear stiffness and damping parameters of an NES on the dynamic response of the pipe. It is concluded that when the nonlinear stiffness of an NES increases to a certain extent, the motion response of the pipe appeared as a saddle-node and Hopf bifurcation, and the vibration amplitude of the pipe is greatly suppressed. For a system with nonlinear motion, NES damping mainly affects the range of the unstable region. For the system without nonlinear motion, the increases in NES damping will reduce the amplitude near the resonance region. Selecting proper parameters of an NES nonlinear stiffness and damping can effectively suppress the vortex-induced vibration of pipes.

Germline and somatic promoter hypermethylation of KLLN has been found in diverse heritable and sporadic cancers, respectively. KLLN has many identified tumor suppressor functions, and when first reported, was thought to be exclusively nuclear. Here, we report on KLLN localization in both the nucleus and cytoplasm and the identification of a putative nuclear export signal (NES) sequence. KLLN overexpression in colon and breast cancer cells showed both nuclear and cytoplasmic presence. Inhibition of the CRM1 export pathway increased nuclear sequestration of KLLN, confirming the prediction of an NES sequence. Point mutations introduced in the predicted NES sequence decreased the strength of the NES and increased the nuclear sequestration of KLLN. Contrary to expectations, the transcription regulation and cellular proliferation functions of KLLN were unaffected by increased KLLN nuclear sequestration. Instead, increased nuclear KLLN correlated with increased nuclear sequestration of TRIM25 and decreased inhibitory phosphorylation of MDM2. Computational analysis of The Cancer Genome Atlas (TCGA) dataset showed positive correlation among KLLN, TRIM25 and MDM2 expression; pathway analysis of the common genes downstream of these three genes revealed protein degradation as one of the top canonical pathways. Together, our observations suggest that CRM1 pathway-based nuclear export of KLLN may impact proteasomal degradation.

Glioblastoma (GBM) remains the leading cause of cancer-related deaths with the lowest five-year survival rates among all of the human cancers. Multiple factors contribute to its poor outcome, including intratumor heterogeneity, along with migratory and invasive capacities of tumour cells. Over the last several years Doublecortin (DCX) has been one of the debatable factors influencing GBM cells’ migration. To resolve DCX’s ambiguous role in GBM cells’ migration, we set to analyse the expression patterns of DCX along with Nestin (NES) and Oligodendrocyte lineage transcription factor 2 (OLIG2) in 17 cases of GBM, using immunohistochemistry, followed by an analysis of single-cell RNA-seq data. Our results showed that only a small subset of DCX positive (DCX+) cells was present in the tumour. Moreover, no particular pattern emerged when analysing DCX+ cells relative position to the tumour margin. By looking into single-cell RNA-seq data, the majority of DCX+ cells were classified as non-cancerous, with a small subset of cells that could be regarded as glioma stem cells. In conclusion, our findings support the notion that glioma cells express DCX; however, there is no clear evidence to prove that DCX participates in GBM cell migration.

En esta presentaci��n se explica c��mo se implementa en una memoria la Tabla de Patrones de la PPU (Picture Processing Unit) de la NES (Nintendo Entertainment System). Los patrones son las teselas en las que se basa la unidad de procesamiento gr��fico de la NES Videos disponibles: Parte A: teor��a https://youtu.be/E4RbOfKB2Js Parte B: implementaci��n en FPGA https://youtu.be/M-aZKPRhV_Y El dise��o de ejemplo est�� disponible: https://github.com/felipe-m/img_rom/tree/master/examples/nesrom/projects/p2_np_tables/smario, {"references":["Machado, Felipe et al, Diseño de sistemas digitales con VHDL http://hdl.handle.net/10115/5700"]}

En esta presentación se explica cómo se implementa en una memoria la Tabla de Patrones de la PPU(Picture Processing Unit) de la NES (Nintendo Entertainment System). Los patrones son las teselas en las que se basa la unidad de procesamiento gráfico de la NES Videos disponibles: Parte A: teoría https://youtu.be/E4RbOfKB2Js Parte B: implementación en FPGA https://youtu.be/M-aZKPRhV_Y El diseño de ejemplo está disponible: https://github.com/felipe-m/img_rom/tree/master/examples/nesrom/projects/p2_np_tables/smario

En esta presentación se explican cómo se implementa en una memoria la Tabla de Nombres de la PPU(Picture Processing Unit) de la NES (Nintendo Entertainment System). La tabla de nombres implementa los gráficos del fondo (background). Se explican también los cálculos que hay que realizar para acceder a las direcciones de memoria de la Tabla de Nombres La unidad gráfica de la NES (la PPU) implementa los gráficos a partir de mapas de teselas. Estos gráficos se utilizaban en las consolas de videojuegos tercera generación, también conocidas como la era de consolas de 8 bits. Videos disponibles: Parte A: teoría https://youtu.be/PBDLakIiw2c Parte B: implementación en FPGA https://youtu.be/Twvdh8xt7a4 El diseño de ejemplo está disponible: https://github.com/felipe-m/img_rom/tree/master/examples/nesrom/projects/nametable/smario La versión 1.1 incluye las imágenes del segundo video.

En esta presentaci��n se explican c��mo se implementa en una memoria la Tabla de Nombres de la PPU (Picture Processing Unit) de la NES (Nintendo Entertainment System). La tabla de nombres implementa los gr��ficos del fondo (background). Se explican tambi��n los c��lculos que hay que realizar para acceder a las direcciones de memoria de la Tabla de Nombres La unidad gr��fica de la NES (la PPU) implementa los gr��ficos a partir de mapas de teselas. Estos gr��ficos se utilizaban en las consolas de videojuegos tercera generaci��n, tambi��n conocidas como la era de consolas de 8 bits. Videos disponibles: Parte A: teor��a https://youtu.be/PBDLakIiw2c Parte B: implementaci��n en FPGA https://youtu.be/Twvdh8xt7a4 El dise��o de ejemplo est�� disponible: https://github.com/felipe-m/img_rom/tree/master/examples/nesrom/projects/nametable/smario La versi��n 1.1 incluye las im��genes del segundo video., {"references":["Machado, Felipe et al, Diseño de sistemas digitales con VHDL http://hdl.handle.net/10115/5700"]}

En esta presentación se explican cómo se realizan imágenes a partir de mapas de teselas.Este sistema se utiliza en las unidades de procesamiento gráfico que utilizan mapas de teselas. Estas unidades se utilizaban en las consolas de videojuegos tercera generación, también conocidas como la era de consolas de 8 bits. Para ello se utiliza como ejemplo la PPU (Picture Processing Unit), que es la unidad de gráficos de la NES (Nintendo Entertainment System). Esta presentación es continuación de la Video disponible: https://youtu.be/sY__uj7bYEo, {"references":["Machado, Felipe et al, Diseño de sistemas digitales con VHDL http://hdl.handle.net/10115/5700"]}

Epstein–Barr virus (EBV), the pathogen of several human malignancies, encodes many proteins required to be transported into the nucleus for viral DNA reproduction and nucleocapsids assembly in the lytic replication cycle. Here, fluorescence microscope, mutation analysis, interspecies heterokaryon assays, co-immunoprecipitation assay, RNA interference, and Western blot were performed to explore the nuclear import mechanism of EBV encoded BLLF2 protein. BLLF2 was shown to be a nucleocytoplasmic shuttling protein neither by a chromosomal region maintenance 1 (CRM1)- nor by a transporter associated with antigen processing (TAP)-dependent pathway. Yet, BLLF2’s two functional nuclear localization signals (NLSs), NLS1 (16KRQALETVPHPQNRGR31) and NLS2 (44RRPRPPVAKRRRFPR58), were identified, whereas the predicted NES was nonfunctional. Finally, BLLF2 was proven to transport into the nucleus via a Ran-dependent and importin β1-dependent pathway. This mechanism may contribute to a more extensive insight into the assembly and synthesis of EBV virions in the nucleus, thus affording a new direction for the treatment of viruses.

3D printing (also known as additive manufacturing) has been developed for more than 30 years. The applications of 3D printing have been increasingly extended to a variety of engineering fields in recent years. The sandwich material with a high strength and overall low density is a kind of artificial material that has been extensively used in various industrial and daily life applications. This paper presents a comprehensive vibration analysis and passive control technique for a cantilevered sandwich beam with an hourglass lattice truss core fabricated with 3D printing technology. The governing equation of the beam is established by using a homogenized model and the Hamilton's principle, from which the natural frequencies are determined. The theoretical model is verified by the results from the existing literature and the finite element analysis. The frequency response of the sandwich beam measured experimentally further validates the proposed model. Subsequently, a non-linear energy sink (NES) is proposed for being employed to passively suppress the vibration of the sandwich beam. A parametric study based on the theoretical model confirms the viability of using NES to effectively control the vibration of the sandwich beam. This work presents a good demonstration of using 3D printing technology for fabricating sandwich beams with a complicated lattice core. More importantly, some guidelines regarding the dynamic analysis of sandwich beams are provided. In addition, the analytical method presented in this work provides a potential means to theoretically explore the advantages of using sandwich beams for energy harvesting in the future.

Fragile X syndrome (FXS) is the most common monogenetic cause of intellectual disability and autism spectrum disorders. Mostly, FXS is caused by transcriptional silencing of the FMR1 gene due to a repeat expansion in the 5′ UTR, and consequently lack of the protein product FMRP. However, in rare cases FXS is caused by other types of variants in the FMR1 gene. We describe a missense variant in the FMR1 gene, identified through whole-exome sequencing, in a boy with intellectual disability and behavioral problems. The variant is located in the FMRP's nuclear export signal (NES). We performed expression and localization studies of the variant in hair roots and HEK293 cells. Our results show normal expression but significant retention of the FMRP in the cells’ nucleus. This finding suggests a possible FMRP reduction at its essential functional sites in the dendrites and the synaptic compartments and possible interference of other cellular processes in the nucleus. Together, this might lead to a FXS phenotype in the boy.

En esta presentación se introducen las unidades de procesamiento gráfico que utilizan mapas de teselas. Estas unidades se utilizaban en las consolas de videojuegos tercera generación, también conocidas como la era de consolas de 8 bits. Para ello se utiliza como ejemplo la PPU (Picture Processing Unit), que es la unidad de gráficos de la NES (Nintendo Entertainment System). Video disponible: https://youtu.be/KCmWw6ioO_o, {"references":["Machado, Felipe et al, Diseño de sistemas digitales con VHDL http://hdl.handle.net/10115/5700"]}

Dans la cellule, les ARN pré messagers contenant des introns sont normalement retenus au noyau par leur interaction avec des facteurs d’épissage. Cependant, les ARN partiellement et non épissés des rétrovirus doivent entrer dans le cytoplasme pour servir de matrice pour la synthèse de certaines protéines telles que Env, Gag et Gag-Pol ainsi que d’ARN génomique qui sera empaqueté dans les nouveaux virions. Un mécanisme post-transcriptionnel utilisé par les lentivirus pour éviter la séquestration nucléaire de ces ARNm dépend d’une protéine virale appelée Rev. Pour assurer sa fonction d’exportation, Rev doit transiter entre le noyau et le cytoplasme et doit aussi pouvoir former des multimères. Par conséquent, Rev est dotée de domaines fonctionnels lui procurant ces habiletés. On retrouve le domaine riche en arginines qui contient le domaine de liaison à l’ARN et le signal de localisation nucléaire (NLS), un second domaine, riche en leucines, porte le signal d’exportation nucléaire (NES) et finalement le domaine de multimérisation. Bien que les protéines Rev du virus de l’immunodéficience humaine de type 1 (VIH-1) et bovine (VIB) aient été caractérisées, aucune étude n’a été réalisée pour la protéine Rev du virus de la maladie de Jembrana (JDV) et très peu sur le virus de l’immunodéficience féline (VIF). Comme les domaines fonctionnels et la voie d’importation des protéines Rev déjà caractérisées sont différents, nous supposons que chaque protéine Rev possède une organisation qui lui est propre et que les mécanismes de transport nucléo-cytoplasmique diffèrent entre les virus. Ce projet a pour objectif de caractériser ces domaines pour la protéine Rev du JDV et ceux du VIF ainsi que les mécanismes permettant leur transport nucléaire. L’utilisation de mutants de la protéine Rev de ces virus couplés à la protéine de fluorescente verte (EGFP) exprimés dans des cellules appropriées et observés par microscopie a permis d’identifier des séquences NLS et NES différentes de celles déjà caractérisées. Le NLS de la protéine Rev du JDV a été identifié et est composé des résidus arginines de la séquence 76-RRPARRPPIRR-87 avec un NoLS composé des mêmes résidus en plus des arginines R74, R103 et R104. Son NES est composé des résidus hydrophobes de la séquence 116-MAELEERFEDLAL-128 et est du type de l’inhibiteur de la protéine kinase (PKI pour « protéine kinase inhibitor »). Pour la protéine Rev du VIF, son NLS est composé des résidus basiques de la séquence 84-KKKRQRRRRKKKAFKK-99. Le NoLS est composé des mêmes acides aminés en plus du résidu K82. De plus, les essais d’importation nucléaires et d’interaction semblent indiquer que les voies d’importation utilisées diffèrent entre les virus et que plusieurs voies peuvent être utilisées. Ces travaux pourront éventuellement servir de base pour identifier de nouvelles cibles thérapeutiques contre les lentivirus., In the cell, pre-messenger RNAs containing introns are normally retained in the nucleus by their interaction with splicing factors. However, the partially and unspliced ​​RNAs of retroviruses must enter the cytoplasm to serve as a template for the synthesis of certain proteins such as Env, Gag and Gag-Pol as well as genomic RNA to be packaged in the new virions. A post-transcriptional mechanism used by lentiviruses to prevent nuclear sequestration of these mRNAs depends on a trans-activator, the viral protein Rev. To ensure its export function, Rev must be able to shuttle between the nucleus and the cytoplasm and to form multimers. As a result, Rev has functional domains that provide these abilities: the arginine-rich domain, which contains the RNA binding domain and the nuclear localization signal (NLS), a second domain, rich in leucine, corresponding to the nuclear export signal (NES) and finally the multimerization domain. Although the Rev proteins of the human and bovine immunodeficiency virus (HIV-1 and BIV respectively) have been characterized, no studies have been performed for the Jembrana disease virus (JDV) Rev protein and very little on the feline immunodeficiency virus (FIV). Since the functional domains and import pathway of the already characterized Rev proteins are different, we assume that each Rev protein has its own organization and that the nucleo-cytoplasmic transport mechanisms differ between viruses. The goal of this project is to characterize these domains for the JDV and FIV Rev proteins as well as to elucidate mechanisms for their nuclear transport. The use of Rev mutants fused to the EGFP expressed in appropriate cells and observed by microscopy has identified NLS and NES sequences that differ from those already characterized. JDV Rev NLS is composed of arginine residues in the 76-RRPARRPPIRR-87 sequence with a NoLS composed of the same residues with the addition of arginine R74, R103 and R104. JDV Rev NES is composed of hydrophobic residues in the 116-MAELEERFEDLAL-128 sequence and is of the protein kinase inhibitor type (PKI). For the FIV Rev protein, its NLS is composed of basic residues in the 84-KKKRQRRRRKKKAFKK-99 sequence. FIV Rev NoLS is composed of the same residues with the addition of the lysine at position 82. In addition, the nuclear import and interaction tests suggest that the import routes used by Rev differ between the different viruses studied and that more than one import pathway may be used. This work could serve as a basis for identifying new therapeutic targets against lentiviruses.

The aim of the study was to further elucidate the feasibility of Sambucus nigra L. fruits in recovery from inflammations and prevention and healing as additional complementary remedy of tumors, as well as the beneficial analgetic and diuretic effects of its flowers. Non-equilibrium energy spectrum (NES) and Differential non-equilibrium energy spectrum (DNES) spectral analyses of hydrogen bonds energies in 1% water extracts were performed. The 1% water extracts of Sambucus nigra L. fruits and flowers were studied. The control sample was with deionized water with chemical purity – 99.99%; pH – 6,0–7,5; electric conductivity – 10 S/cm. Increased average energy of hydrogen bonds compared to deionized water was found in both types of extracts as follows: ∆Efr=-0.0139±0.0007 eV for fruits and ∆Efl =-0.0079±0.0004 eV for flowers. The biggest local extremums of hydrogen bond energies in both types of extracts were measured at (-0.1112 eV) (λ=11.15 μm)(ṽ=897 cm-1), (-0.1212 eV) (λ=10.23 μm)( ṽ=978 cm-1 )and (-0.1387 eV) (λ=8.95 μm)(ṽ=1117 cm-1) i. These extremums are identical with extremums observed in other studies where they have been found to correspond to analgetic and diuretic, anti-inflammatory and anti-tumor effects.

Writing center tutors strive to facilitate participation from student writers, particularly student writers who are not native speakers of the conference language. This study investigated one way that tutors might better understand student writers’ intent to contribute a substantial turn at talk and thus better understand when they might make way for student writers’ active participation. This study examined four minimal responses (MRs)—mmhm, uhhuh, yeah, and ok—at the beginning of student writers’ turns at talk. It differentiated between MRs that were free standing, constituting the entire turn and suggesting passive recipiency, and MRs that were not free standing, suggesting speakership incipiency. Importantly, the study differentiated between the MRs of native English speakers (NESs) and non-native English speakers (NNES). NNESs used freestanding mmhm far more than NESs, suggesting that the NNESs may have extended the use of mmhm to a greater array of discourse contexts. NNESs used free-standing yeah far more frequently than they did non-free-standing yeah, suggesting that yeah would not have been a reliable signal for tutors that NNESs would extend their turns at talk. This study also found that both NESs and NNESs used ok to signal not only consideration of but also agreement with tutors’ evaluations or acceptance of tutors’ advice about lower-order concerns. Understanding how MRs vary from passive recipiency to speakership incipiency might help tutors better understand student writers’ intent to contribute a substantial turn and thus indicate when tutors might wait for student writers’ participation.

[ES] Este trabajo trata de la explicación de la creación de gráficos en dispositivos electrónicos a finales de los años 70 y principios de los años 80, la aplicación de dichos gráficos y/o la creación de directrices teniendo en cuenta las limitaciones técnicas de estos con el propósito de crear gráficos fieles a los productos mencionados. En este trabajo, se detallarán las limitaciones de cada dispositivo, las herramientas y técnicas usadas, la aplicación de fundamentos de arte y animación de los gráficos, los obstáculos que aparecen en este ámbito, la solución a estos problemas, la explicación del valor artístico y el uso aplicado de los conocimientos y prácticas de este proyecto. Los dispositivos mencionados son: Apple II, Atari 2600/VCS, Commmodore 64 y NES/Famicom., [EN] This work is about the explanation of the creation of graphics in electronic devices during the late 70s and early 80s, the application of said graphics and/or the creation of guidelines taking into account the technical limitations of these with the purpose of creating graphics loyal to the mentioned products. In this work shall be detailed the limitations of each device, the tools and techniques used, the application of art and animation fundamentals on the graphics, the obstacles that appear in this field, the solution to these problems, the explanation of the artistic value and the applied use of the knowledge and exercise of this project. The mentioned devices are: Apple II, Atari 2600/VCS, Commmodore 64 and NES/Famicom.

Spectral analyses of 1% water extracts of fresh and dry Hypericum perforatum L. and 1% dry H. perforatum with colloidal nano silver (NSPs) 30 ppm were conducted. The nano silver is standardised and patented by the Swiss company Evodrop. Non-equliblrium energy spectrum (NES) and Differential non-equliblrium energy spectrum (DNES) methods were used for the spectral analysis. A comparative analysis of 1% extracts of fresh and dry H. perforatum was performed in order to determine the local extremums for effects of nerve tissue conductivity at (-0.1112) eV, anti-inflammatory (-0.1212) eV, anti-tumor effects (-0.1387) eV. The results showed stimulating effect on the nervous system and improvement of nerve conduction (local extremums E=-0.1112 eV)(?=11.15 ?m) (?=897 cm-1), as well as anti-inflammatory effect (E = -0.1212 eV)( ?=10.23 ?m) (?=978 cm-1) and inhibition of development of tumor cells at a molecular level (E=-0.1387 eV) (?=8.95 ?m) (?=1117 cm-1). It was found that clusters of 16 and 15 water molecules are formed in the water herbal extracts of fresh H. perforatum and of dry H. perforatum with AgNPs 30 ppm. The fresh plant showed better results then the dry one. The addition of colloidal nano silver 30 ppm led to better results of the drug.

The nuclear export receptor CRM1 (XPO1) recognizes and binds specific sequence motifs termed nuclear export signals (NESs) in cargo proteins. About 200 NES motifs have been identified, but over a thousand human proteins are potential CRM1 cargos, and most of their NESs remain to be identified. On the other hand, the interaction of NES peptides with the &ldquo, NES-binding groove&rdquo, of CRM1 was studied in detail using structural and biochemical analyses, but a better understanding of CRM1 function requires further investigation of how the results from these in vitro studies translate into actual NES export in a cellular context. Here we show that a simple cellular assay, based on a recently described reporter (SRVB/A), can be applied to identify novel potential NESs motifs, and to obtain relevant information on different aspects of CRM1-mediated NES export. Using cellular assays, we first map 19 new sequence motifs with nuclear export activity in 14 cancer-related proteins that are potential CRM1 cargos. Next, we investigate the effect of mutations in individual NES-binding groove residues, providing further insight into CRM1-mediated NES export. Finally, we extend the search for CRM1-dependent NESs to a recently uncovered, but potentially vast, set of small proteins called micropeptides. By doing so, we report the first NES-harboring human micropeptides.

El objetivo es investigar sobre las capacidades y las limitaciones que tiene la Nintendo Entertainment System y las herramientas disponibles, actualmente, para desarrollar un videojuego completo para ella, íntegro en ensamblador. La finalidad del proyecto es mejorar las capacidades del alumno y crear una documentación referencia para futuros desarrolladores que supla las carencias de la disponible actualmente. Aprender a programar sobre consolas retro ayuda al alumno a profundizar en sus conocimientos de ingeniería debido a las limitaciones de estas consolas. Además el diseño del videojuego ayuda al alumno a mejorar su organización de cara a proyectos laborales. Se abordará la investigación realizada, el diseño y el desarrollo del videojuego. Por último, este tipo de proyectos son ideales en el portfolio del alumno, ya que no todos los alumnos abordan este tipo de temas a la hora de realizar el Trabajo de Fin de Grado y cabe la posibilidad que las empresas lleguen a interesarse por éstos.

International audience; In order to have the targeted energy transfer in the acoustic domain, R. Bellet proposed a non-linear vibro-acoustic absorber constituted by a thin viscoelastic circular membrane coupled to the first resonance mode of an open tube through a coupling box and excited by an acoustic field provided by a loudspeaker. The membrane damping in the Bellet?s model is described in term of linear and quadratic viscous-like damping. The damping coefficient were obtained using identification in the linear case. The comparison between experiments and computation revealed that this description was not sufficient to capture all the feature of the very rich dynamics of this nonlinear system. We concluded a single value of the damping coefficient independent of the amplitude of the excitation, was the origin of this discrepancy. We decided to estimate the damping coefficient using the Bellet?s model in both linear and non-linear domains. For the estimation of this coefficient, a series of experimental tests was realized, in which the amplitude of the sinusoidal voltage applied at the terminal of the loudspeaker changed and the frequency is varied for each input level. This experiment lets the measure of the pressure in the middle of the tube by a microphone and the displacement of the membrane in its center by a displacement sensor. Moreover, following these tests we have developed a MATHEMATICA code that allows the nonlinear identification of the damping coefficient of the NES using the harmonic balance method. We present the estimated variation of the damping coefficient as a function of the excitation amplitude, wherein a sudden increase of the dissipation during the energy pumping is clearly observed. This clearly indicates that some dissipative phenomenon neglected in our model must be accounted for a better description of energy pumping over its completely functioning range.

En este proyecto se experimentó con algoritmos de aprendizaje por refuerzo jugando Donkey Kong, Ice Climber, Kung Fu, Super Mario Bros y Metroid de la consola NES. El algoritmo DQN y sus variantes como Doble DQN y Dueling DQN fueron usados para experimentar con la representación de estados mediante la RAM. Se proponen algunas estrategias para reducir la dimensionalidad de los estados y las acciones. También se proponen funciones de recompensa para crear un agente fácil de entrenar con pocos recursos computacionales. Se probaron dos maneras de reducir la dimensión de la RAM, el mapa de la RAM funcionó bien sólo en fase de entrenamiento mientras que el método de los bytes activados consiguió mejores resultados. In this project, reinforcement learning algorithms are explored to play the games Donkey Kong, Ice Climber, Kung Fu, Super Mario Bros, and Metroid from the NES console. The Deep-Q learning algorithm is used to experiment with the RAM representation of the state. DQN algorithm extensions as Double DQN and Dueling Network Architectures are explored too. Some simple strategies to reduce the state-space and action-space are proposed in addition to reward functions to create an easy to train agent with low computational resources. Two ways to reduce the RAM representation of the state were tested, RAM map method worked well just in the training phase meanwhile the activated bytes method get better results also in gameplay. En aquest projecte es va experimentar amb algorismes d'aprenentatge per reforç jugant Donkey Kong, Hissi Climber, Kung Fu, Super Mario Bros i Metroid de la consola NES. L'algorisme DQN i les seves variants com a Doble DQN i Dueling DQN van ser usats per a experimentar amb la representació d'estats mitjançant la RAM. Es proposen algunes estratègies per a reduir la dimensionalitat dels estats i les accions. També es proposen funcions de recompensa per a crear un agent fàcil d'entrenar amb pocs recursos computacionals. Es van provar dues maneres de reduir la dimensió de la RAM, el mapa de la RAM va funcionar bé només en fase d'entrenament mentre que el mètode dels bytes activats va aconseguir millors resultats.

Made available in DSpace on 2020-12-12T01:17:41Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-05-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Replication protein A (RPA), a heterotrimeric complex, is the major single-stranded DNA binding protein in eukaryotes. Recently, we characterized RPA from Trypanosoma cruzi, showing that it is involved in DNA replication and DNA damage response in this organism. Better efficiency in differentiation from epimastigote to metacyclic trypomastigote forms was observed in TcRPA-2 subunit heterozygous knockout cells, suggesting that RPA is involved in this process. Here, we show that RPA cellular localization changes during the T. cruzi life cycle, with RPA being detected only in the cytoplasm of the metacyclic and bloodstream trypomastigotes. We also identify a nuclear export signal (NES) in the trypanosomatid RPA-2 subunit. Mutations in the negatively charged residues of RPA-2 NES impair the differentiation process, suggesting that RPA exportation affects parasite differentiation into infective forms. Laboratório de Ciclo Celular Instituto Butantan Center of Toxins Immune Response and Cell Signaling (CeTICS) Instituto Butantan Departamento de Física e Biofísica Instituto de Biociências Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP) Laboratorie de Biologie et Pharmacologie Appliquée Ecole Normale Supérieure Paris-Saclay Instituto Carlos Chagas Fiocruz Laboratório de Parasitologia Instituto Butantan Departamento de Física e Biofísica Instituto de Biociências Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP) FAPESP: 2013/07467-1 FAPESP: 2014/02978-0 FAPESP: 2016/50050-2 FAPESP: 2018/12364-0 FAPESP: 2018/14432-3 FAPESP: 2018/21785-0 CNPq: 306199/2018-1 CAPES: 88881.171895/2018-01

Neuronal Per-Arnt-Sim (PAS) domain-containing protein 4 (NPAS4) is a basic helix-loop-helix (bHLH)-PAS transcription factor first discovered in neurons in the neuronal layer of the mammalian hippocampus and later discovered in pancreatic β-cells. NPAS4 has been proposed as a therapeutic target not only for depression and neurodegenerative diseases associated with synaptic dysfunction but also for type 2 diabetes and pancreas transplantation. The ability of bHLH-PAS proteins to fulfil their function depends on their intracellular trafficking, which is regulated by specific sequences, i.e. the nuclear localization signal (NLS) and the nuclear export signal (NES). However, until now, no study examining the subcellular localization signals of NPAS4 has been published. We show here that Rattus norvegicus NPAS4 was not uniformly localized in the nuclei of COS-7 and N2a cells 24 h after transfection. Additionally, cytoplasmic localization of NPAS4 was leptomycin B-sensitive. We demonstrate that NPAS4 possesses a unique arrangement of localization signals. Its bHLH domain contains an overlapping NLS and NES. We observed that its PAS-2 domain contains an NLS, an NES, and a second, proximally located, putative NLS. Moreover, the C terminus of NPAS4 contains two active NESs that overlap with a putative NLS. Our data indicate that glucose concentration could be one of the factors influencing NPAS4 localization. The presence of multiple localization signals and the differentiated localization of NPAS4 suggest a precise, multifactor-dependent regulation of NPAS4 trafficking, potentially crucial for its ability to act as a cellular stress sensor and transcription factor.

DEAD-box helicase 3, X-linked (DDX3X) regulates the retinoic acid-inducible gene I (RIG-I)-like receptor (RLR)-mediated antiviral response, but can also be a host factor contributing to the replication of viruses of significance to human health, such as human immunodeficiency virus type 1 (HIV-1). These roles are mediated in part through its ability to actively shuttle between the nucleus and the cytoplasm to modulate gene expression, although the trafficking mechanisms, and impact thereof on immune signaling and viral infection, are incompletely defined. We confirm that DDX3X nuclear export is mediated by the nuclear transporter exportin-1/CRM1, dependent on an N-terminal, leucine-rich nuclear export signal (NES) and the monomeric guanine nucleotide binding protein Ran in activated GTP-bound form. Transcriptome profiling and ELISA show that exportin-1-dependent export of DDX3X to the cytoplasm strongly impacts IFN-&beta, production and the upregulation of immune genes in response to infection. That this is key to DDX3X&rsquo, s antiviral role was indicated by enhanced infection by human parainfluenza virus-3 (hPIV-3)/elevated virus production when the DDX3X NES was inactivated. Our results highlight a link between nucleocytoplasmic distribution of DDX3X and its role in antiviral immunity, with strong relevance to hPIV-3, as well as other viruses such as HIV-1.

The bHLH-PAS (basic helix-loop-helix/ Period-ARNT-Single minded) proteins are a family of transcriptional regulators commonly occurring in living organisms. bHLH-PAS members act as intracellular and extracellular “signals” sensors, initiating response to endo- and exogenous signals, including toxins, redox potential, and light. The activity of these proteins as transcription factors depends on nucleocytoplasmic shuttling: the signal received in the cytoplasm has to be transduced, via translocation, to the nucleus. It leads to the activation of transcription of particular genes and determines the cell response to different stimuli. In this review, we aim to present the current state of knowledge concerning signals that affect shuttling of bHLH-PAS transcription factors. We summarize experimentally verified and published nuclear localization signals/nuclear export signals (NLSs/NESs) in the context of performed in silico predictions. We have used most of the available NLS/NES predictors. Importantly, all our results confirm the existence of a complex system responsible for protein localization regulation that involves many localization signals, which activity has to be precisely controlled. We conclude that the current stage of knowledge in this area is still not complete and for most of bHLH-PAS proteins an experimental verification of the activity of further NLS/NES is needed.

International audience; A nonlinear magnetic vibration absorber is presented and used to control vibration of a three-storey structure. A distinctive feature of the absorber concerns its versatility for tuning the linear and nonlinear stiffness coefficients, depending on simple geometric design parameters such as the distance between fixed magnets and the moving one. In particular, the absorber can be set either as a nonlinear tuned vibration absorber, a nonlinear energy sink, or a bistable tuned vibration absorber, according to whether the linear stiffness term is positive, vanishing, or negative. The response of the primary structure and the vibration mitigation are investigated in the cases of impulsive shock, free vibration with imposed initial displacement, and single frequency excitation. Significant reductions of the primary structure vibrations are obtained for the three cases investigated, showing the ability of using a vibration absorber only relying on magnetic forces for passive control. The detailed comparisons of the absorbers performance show that, in this case study, no general guidelines can be easily deduced for selecting one of the three tunings for a nonlinear absorber. Depending on the excitation, the vibratory levels, and the frequency content of the excitation, the three configurations show advantages and drawbacks that are discussed.

Master's thesis in Computer science Music generation using deep learning is a widely studied field. This thesis focuses on music generation in a constrained and novel environment; retro video game music. The constraints imposed by the environment creates many unique challenges for the generation of musical compositions. In addition, the dataset consists of multi-instrument music, which is rarely studied due to its complexity. An extension to an existing architecture; the Biaxial RNN is presented in order to extend its capabilities to allow for generating multi-instrument arrangements. The resulting implementation is somewhat successful at fulfilling the proposed solution, although one component could not be implemented within the time limit. The result is not pleasant music, but it does give a view into the complex process of multi-instrumental music generation.

Упитанность сомов Hybryd Clarias sp. выращенных в садковых хозяйствах реки Кай варьирует от 0.14 до 1.76 при среднем значении 1.12±0.02. Скелетные мышцы содержат воды 70.17±0.27%, сухого вещества 30.10±0.40%, белка 22.63±0.57%, жира 2.90±0.60%, золы 1.70±0.03%, БЭВ 2.87±0.72%. Показано, что мышечная ткань особей Hybryd Clarias sp. содержит большее количество воды, жира и минеральных веществ и меньшее количество сухого вещества и белка по сравнению с популяциями Clarias batrachus. Выдвинуто предположение, что хозяйственные условия выращивания (нарушение технологии кормления и содержания) Hybryd Clarias sp. в садковых хозяйствах Южного Вьетнама негативно влияют на массу и упитанность особей, а также химический состав скелетных мышц., Fat catfish Hybryd Clarias sp. grown in cage farms of the Kai River varies from 0.14 to 1.76 with an average value of 1.12±0.02. Skeletal muscles contain water 70.17±0.27%, dry matter 30.10±0.40%, protein 22.63±0.57%, fat 2.90±0.60%, ash 1.70±0.03%, NES 2.87±0.72%. It was shown that the muscle tissue of individuals of Hybryd Clarias sp. contains more water, fat and minerals and less dry matter and protein than Clarias batrachus populations. It has been suggested that the economic conditions of cultivation (violation of feeding and keeping technology) Hybryd Clarias sp. in cage farms in South Vietnam negatively affect the weight and fatness of individuals, as well as the chemical composition of skeletal muscles.

We start by summarizing noise and vibration problematics.Then available modeling tools are presented: materials and sub-structures, absorption, isolation, damping and encapsulation.Concerning vibrations, emphasis is put on structural damping: decoupling pads, viscous damping, Tuned mass dampers and Non-linear energy sinks. These solutions are compared in terms of pro and cons.

Detta examensarbete skapades med målet att utforska musik från Nintendo Entertainment System–spel (NES) med ett musikteoretiskt och musikterminologiskt perspektiv. Det analyserade stycket är Title Theme från The Legend of Zelda (1988). Bakgrundskapitlet tar upp de verktyg kompositörerna hade att arbeta med som NES:s ljudkort och loopar. Plankning och transkription diskuteras och tidigare forskning i området tv-spel synliggörs. Syftet är att skapa en visuell analys som kan användas för att arrangera om originalmusiken från spelen. Genom plankning och analys har jag producerat ett nytt arrangemang. Resultatet presenteras som noter på min plankning, analys och arrangemang med förklaringar över originalets uppbyggnad. Arbetet har gett insikt i hur min skolning i musik kan vara hämmande för att arrangera som programmerarna gjorde förr, men också vilka verktyg de hade att arbeta med. Med arbetet har jag insett hur hjälpsam en tydlig analys av ett verk kan vara för arrangering.

Entrepreneurship is widely argued to be critical for economic development and alleviating extreme poverty. However, entrepreneurship research in sub-Saharan Africa has not received much attention over the last few decades possibly due to a lack of sufficient resources. It is becoming increasingly important as Africa, especially sub-Saharan Africa, is developing rapidly and moving from a resource-based economy to one of innovation and progress. Using data from the Global Entrepreneurship Monitor (GEM), this paper discusses the opinions of national expert informants in Angola, Madagascar, Mozambique and South Africa and looks at the factors which are possibly hindering and inhibiting entrepreneurial development. The results indicate that there are four main inhibitors ranging from lack of access to finance, government policies, regulations and practices for entrepreneurs and the poor levels of entrepreneurship education. Some recommendations are made as to what can be done to assist in promoting economic development.

We start by summarizing noise and vibration problematics.Then available modeling tools are presented: materials and sub-structures, absorption, isolation, damping and encapsulation.Concerning vibrations, emphasis is put on structural damping: decoupling pads, viscous damping, Tuned mass dampers and Non-linear energy sinks. These solutions are compared in terms of pro and cons.

[ES] KMT2D es una metiltransferasa que se encarga de transferir un grupo metilo a la histona H3, lo que está relacionado con la activación génica. Esta proteína es esencial en el desarrollo embrionario temprano y, además, se expresa también en numerosos tejidos adultos. Las mutaciones en el gen que codifica para la síntesis de KMT2D en la línea germinal están relacionadas con el síndrome de Kabuki, una enfermedad congénita rara. También se ha visto que las mutaciones somáticas están relacionadas con la formación de tumores, siendo KMT2D una de las proteínas más frecuentemente mutadas en el cáncer. Por su actividad metiltransferasa de histonas, se sabe que KMT2D desarrolla su función en el núcleo. Partiendo de la hipótesis inicial de que KMT2D puede tener señales de transporte nucleocitoplásmico funcionales, se realizaron análisis bioinformáticos para predecir tanto posibles señales de localización nuclear (NLS) como señales de exportación nuclear (NES) en su secuencia. Los ensayos experimentales revelaron que KMT2D tiene una NLS funcional y una NES débil. De este modo, si se identifican mutaciones relacionadas con patologías en las regiones en las que se encuentran estas señales, se podría investigar la relación del transporte alterado de KMT2D con las diferentes patologías.

As a transcription factor, localization to the nucleus and the recruitment of co-factors to regulate gene transcription is essential. Nuclear localization and nucleosome remodeling and histone deacetylase (NuRD) complex binding are required for the zinc finger transcription factor CASZ1 to function as neuroblastoma (NB) tumor suppressor. However, the critical amino acids (AAs) that are required for CASZ1 interaction with NuRD complex and the regulation of CASZ1 subcellular localization have not been characterized. Through alanine scanning, immunofluorescence cell staining and co-immunoprecipitation we define a critical region at the CASZ1 N-terminus (AA23-40) that mediates the CASZ1b nuclear localization and NuRD interaction. Furthermore, we identify a nuclear export signal (NES) at the N-terminus (AA176-192) that contributes to CASZ1 nuclear-cytoplasmic shuttling in a chromosomal maintenance 1 (CRM1)-dependent manner. An analysis of CASZ1 protein expression in a primary neuroblastoma tissue microarray shows that high nuclear CASZ1 staining is detected in tumors from NB patients with good prognoses. In contrast, cytoplasmic-restricted CASZ1 staining or low nuclear CASZ1 staining is found in tumors from patients with poor prognoses. These findings provide insight into mechanisms by which CASZ1 regulates transcription, and suggests that regulation of CASZ1 subcellular localization may impact its function in normal development and pathologic conditions such as neuroblastoma tumorigenesis.

Maeda H Mohammad,1 Ahmed M Al-Shammari,1 Ahmad Adnan Al-Juboory,2 Nahi Y Yaseen1 1Experimental Therapy Department, Iraqi Center of Cancer and Medical Genetic Research, Al-Mustansiriyah University, 2Department of Surgery, Neuroscience Hospital, Baghdad, Iraq Abstract: The in vitro isolation, identification, differentiation, and neurogenesis characterization of the sources of mesenchymal stem cells (MSCs) were investigated to produce two types of cells in culture: neural cells and neural stem cells (NSCs). These types of stem cells were used as successful sources for the further treatment of central nervous system defects and injuries. The mouse bone marrow MSCs were used as the source of the stem cells in this study. β-Mercaptoethanol (BME) was used as the main inducer of the neurogenesis pathway to induce neural cells and to identify NSCs. Three types of neural markers were used: nestin as the immaturation stage marker, neurofilament light chain as the early neural marker, and microtubule-associated protein 2 as the maturation marker through different time intervals in the neurogenesis process starting from the MSCs, (as undifferentiated cells), NSCs, production stages, and toward neuron cells (as differentiated cells). The results of different exposure times to BME of the neural markers analysis done by immunocytochemistry and real time-polymerase chain reaction helped us to identify the exact timing for the neural stemness state. The results showed that the best exposure time that may be used for the production of NSCs was 6 hours. The best maintenance media for NSCs were also identified. Furthermore, we optimized exposure to BME with different times and concentrations, which could be an interesting way to modulate specific neuronal differentiation and obtain autologous neuronal phenotypes. This study was able to characterize NSCs in culture under differentiation for neurogenesis in the pathway of the neural differentiation process by studying the expressed neural genes and the ability to maintain these NSCs in culture for further differentiation in thousands of functional neurons for the treatment of brain and spinal cord injuries and defects. Keywords: mesenchymal stem cells, neural stem cells, NES, NF-L, MAP-2

Detta examensarbete skapades med målet att undersöka skillnader i preferensen gällande arpeggios i NES- och Famicom-musik baserat på kultur. Bakgrundskapitlet tar upp tidigare forskning inom tangentiella ämnen, samt beskriver arpeggios och hur NES- och Famicom-konsolerna är uppbyggda. En kvantitativ metod utvecklas sedan, och problematiseras. Detta följs av en beskrivning av det förarbete som gjordes, och skapandet av de två artefakter som användes. En enkät genomfördes efter förarbetet, och resultatet av denna analyseras därefter i rapporten. Resultatet visar på att det finns vissa skillnader i preferens mellan svenska och japanska lyssnare. Denna slutsats, samt resterande delar av rapporten diskuteras sedan och till sist skrivs det om framtida arbete, så som att en plug-in möjligtvis skulle kunna skapas efter detta arbete har gjorts. Denna plugin skulle göra det lättare att få musik att låta mer europeisk eller japansk.

We start by summarizing noise and vibration problematics.Then available modeling tools are presented: materials and sub-structures, absorption, isolation, damping and encapsulation.Concerning vibrations, emphasis is put on structural damping: decoupling pads, viscous damping, Tuned mass dampers and Non-linear energy sinks. These solutions are compared in terms of pro and cons.

Recent studies have demonstrated the interference of nucleocytoplasmic trafficking with the establishment and maintenance of various cancers. Nucleocytoplasmic transport is highly regulated and coordinated, involving different nuclear transport factors or receptors, importins and exportins, that mediate cargo transport from the cytoplasm into the nucleus or the other way round, respectively. The exportin CRM1 (Chromosome region maintenance 1) exports a plethora of different protein cargoes and ribonucleoprotein complexes. Structural and biochemical analyses have enabled the deduction of individual steps of the CRM1 transport cycle. In addition, CRM1 turned out to be a valid target for anticancer drugs as it exports numerous proto-oncoproteins and tumor suppressors. Clearly, detailed understanding of the flexibility, regulatory features and cooperative binding properties of CRM1 for Ran and cargo is a prerequisite for the design of highly effective drugs. The first compound found to inhibit CRM1-dependent nuclear export was the natural drug Leptomycin B (LMB), which blocks export by competitively interacting with a highly conserved cleft on CRM1 required for nuclear export signal recognition. Clinical studies revealed serious side effects of LMB, leading to a search for alternative natural and synthetic drugs and hence a multitude of novel therapeutics. The present review examines recent progress in understanding the binding mode of natural and synthetic compounds and their inhibitory effects.

L’objectif de cette thèse est de contrôler passivement une instabilité dynamique appliquée au flottement d’un profil aéroélastique à l’aide de différents types d’Amortisseurs à Masse Accordés (AMA). Un profil 2D appelé Section Typique est utilisé tout au long de l’étude. En première partie, une étude comparative de trois modèles mathématiques d’interaction fluide/structure appliqués à la Section Typique (Theodorsen, LUVLM et UVLM) met en valeur les forces et faiblesses de chacun. Le banc d’essai aéroélastique en soufflerie, utilisé par la suite, est présenté puis identifié avec et sans vent (GVT). En deuxième partie, les calculs des vitesses critiques de Divergence, d’Inversion des Gouvernes et de Flottement sont automatisés avec le modèle Theodorsen afin de réaliser une étude paramétrique du banc d’essai et mettre en lumière les variables de conception les plus influentes. L’analyse modale présente différentes bifurcations liées au changement soudain du mode instable. Ensuite, le même algorithme est utilisé afin d’analyser la suppression du flottement à l’aide de trois géométries d’AMAs linéaires. La dernière partie présente l’étude expérimentale et numérique d’un AMA non linéaire de type Nonlinear Energy Sink (NES). La singularité de cette configuration est d’utiliser le volet en tant qu’amortisseur et ainsi, ne pas ajouter de masse (FSI-VA). En soufflerie, six comportements non linéaires sous-critiques (en deçà de la vitesse de flottement dans la configuration linéaire) sont observés, identifiés et analysés : cinq Cycles Limites d’Oscillations (LCO) et un battement non linéaire chaotique. The aim of this thesis is to passively control a dynamic instability applied to an aeroelastic profile’s flutter using different types of Tuned Mass Dampers (TMD). A 2D profile called Typical Section is used throughout the study. In the first part, a comparative study of three mathematical models of fluid-structure interaction applied to the Typical Section (Theodorsen, LUVLM and UVLM) highlights the strengths and weaknesses of each code. The aeroelastic test bench, used subsequently, is presented and identified with and without wind (Ground Vibration Test, GVT). In the second part, critical velocities computations (Divergence, Control Surface Reversal and flutter) are automated while using the Theodorsen model in order to carry out the test bench parametrical study to highlight most influential variables. The modal analysis presents different bifurcations linked to the sudden change of the unstable mode. The last part presents the experimental and numerical studies of a nonlinear TMD called Nonlinear Energy Sink (NES). The uniqueness of this configuration consists in recycling flap’s vibrations as a flutter damper and thus, get a zero added mass. A nonlinear restoring force can be achieved by a highly nonlinear mechanism. The nonlinear structural behavior is derived analyticaly and is in good agreement with experimental torsion tests. In the wind tunnel, six subcritical nonlinear behaviors (below the flutter velocity in the linear configuration case) are observed, identified and then analyzed : five Limit Cycle Oscillations (LCO) and a chaotic nonlinear beating.

Nuclear export receptor CRM1 binds highly variable nuclear export signals (NESs) in hundreds of different cargoes. Previously we have shown that CRM1 binds NESs in both polypeptide orientations (Fung et al., 2015). Here, we show crystal structures of CRM1 bound to eight additional NESs which reveal diverse conformations that range from loop-like to all-helix, which occupy different extents of the invariant NES-binding groove. Analysis of all NES structures show 5-6 distinct backbone conformations where the only conserved secondary structural element is one turn of helix that binds the central portion of the CRM1 groove. All NESs also participate in main chain hydrogen bonding with human CRM1 Lys568 side chain, which acts as a specificity filter that prevents binding of non-NES peptides. The large conformational range of NES backbones explains the lack of a fixed pattern for its 3-5 hydrophobic anchor residues, which in turn explains the large array of peptide sequences that can function as NESs. DOI: http://dx.doi.org/10.7554/eLife.23961.001

To reduce the vibrations seen by a product, three main families of solutions (physics) exist: to realize a decoupling (decoupling pads), to thermally dissipate energy (viscous damping) or to mechanically capture energy (Tuned mass dampers; Non-linear energy sinks).We will briefly present these different possibilities, their drawbacks and advantages, their scope of use, and the main points entering their dimensioning.Emphasis will be put on viscous damping and NES since experimental results will be presented and compared to simulation.

International audience; We try to enlighten the time multi-scale energy exchange between a main oscillator of Bouc-Wen type and a nonlinear energy sink. A general methodology is presented to detect the invariant manifold of the system at fast time scale and then to search equilibrium points and fold singularities at the slow time scale. A numerical example is enclosed to show the application of the coupled nonlinear energy sink in passive control of Bouc-Wen type main structures.

ZO-2 is a cytoplasmic protein of tight junctions (TJs). Here, we describe ZO-2 involvement in the formation of the apical junctional complex during early development and in TJ biogenesis in epithelial cultured cells. ZO-2 acts as a scaffold for the polymerization of claudins at TJs and plays a unique role in the blood–testis barrier, as well as at TJs of the human liver and the inner ear. ZO-2 movement between the cytoplasm and nucleus is regulated by nuclear localization and exportation signals and post-translation modifications, while ZO-2 arrival at the cell border is triggered by activation of calcium sensing receptors and corresponding downstream signaling. Depending on its location, ZO-2 associates with junctional proteins and the actomyosin cytoskeleton or a variety of nuclear proteins, playing a role as a transcriptional repressor that leads to inhibition of cell proliferation and transformation. ZO-2 regulates cell architecture through modulation of Rho proteins and its absence induces hypertrophy due to inactivation of the Hippo pathway and activation of mTOR and S6K. The interaction of ZO-2 with viral oncoproteins and kinases and its silencing in diverse carcinomas reinforce the view of ZO-2 as a tumor regulator protein.

Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-27T11:29:49Z No. of bitstreams: 0 Made available in DSpace on 2014-05-27T11:29:49Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-07-01 The objective was to evaluate reproductive tract development (ovary and uterus) and onset of puberty in two lines of Nellore heifers (Bos indicus) selected for postweaning weight. A total of 123 heifers, including 46 from the control Nellore line (NeC) and 77 from the selection Nellore line (NeS) were used. Every 18 to 21 days from 12 to 24 months of age, average ovarian area (OVA), endometrial thickness (ETh), and diameter of the largest follicle in each ovary were evaluated (using transrectal ultrasonography), and body weight, hip height, and body condition score were measured. There were no differences between NeS and NeC heifers for ETh or OVA (P < 0.05). Genetic selection for higher postweaning weight had no negative influence on the onset of puberty, with 52% and 48% of NeC and NeS heifers, respectively, pubertal at 24 months of age (P = 0.49). Heifers that reached puberty at the end of the study were heavier (NeC, 296.9 vs. 276.7 kg; NeS, 343.5 vs. 327.9 kg; P < 0.01) and younger (NeC, 23.4 vs. 24.2 mo; NeS, 22.7 vs. 24.0 months; P < 0.01) than those that did not. Furthermore, heifers that were heavier at weaning reached puberty earlier. Pubertal heifers had a greater OVA (4.15 vs. 3.14 cm2; P < 0.01) and ETh (12.15 vs. 9.93 mm; P < 0.01) than nonpubertal heifers. Taken together, OVA and ETh had positive effects (P < 0.01) on the onset of puberty and were suitable indicator traits of heifer sexual precocity in pasture management systems. However, selection for weight did not alter ovarian or endometrial development, or manifestation of puberty at 24 months of age. Among the growth traits studied, weaning weight and weight at puberty had significant positive effects on manifestation of first estrus. © 2013 Elsevier Inc. Centro APTA Bovinos de Corte IZ-APTA, Sertãozinho, São Paulo Department of Animal Reproduction FCAV-UNESP, Jaboticabal, São Paulo Department of Pharmacology IBB-UNESP, Botucatu, Sao Paulo Department of Animal Breeding and Nutrition FMVZ-UNESP, Botucatu, São Paulo Department of Animal Reproduction FCAV-UNESP, Jaboticabal, São Paulo Department of Pharmacology IBB-UNESP, Botucatu, Sao Paulo Department of Animal Breeding and Nutrition FMVZ-UNESP, Botucatu, São Paulo

The myocardin-related transcription factors (MRTF-A and MRTF-B) regulate cytoskeletal genes through their partner transcription factor SRF. The MRTFs bind G-actin, and signal-regulated changes in cellular G-actin concentration control their nuclear accumulation. The MRTFs also undergo Rho- and ERK-dependent phosphorylation, but the function of MRTF phosphorylation, and the elements and signals involved in MRTF-A nuclear export are largely unexplored. We show that Rho-dependent MRTF-A phosphorylation reflects relief from an inhibitory function of nuclear actin. We map multiple sites of serum-induced phosphorylation, most of which are S/T-P motifs and show that S/T-P phosphorylation is required for transcriptional activation. ERK-mediated S98 phosphorylation inhibits assembly of G-actin complexes on the MRTF-A regulatory RPEL domain, promoting nuclear import. In contrast, S33 phosphorylation potentiates the activity of an autonomous Crm1-dependent N-terminal NES, which cooperates with five other NES elements to exclude MRTF-A from the nucleus. Phosphorylation thus plays positive and negative roles in the regulation of MRTF-A. DOI: http://dx.doi.org/10.7554/eLife.15460.001