University, Diss., 2005--Hamburg.

Orientador: Lea Rodrigues Simioni / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-11T09:48:32Z (GMT). No. of bitstreams: 1 Abreu_ValdemirAparecidode_D.pdf: 2741527 bytes, checksum: 8d40cb65f4500adbbbd0961f047f41b1 (MD5) Previous issue date: 2008 / Resumo: Neste trabalho, foi estudada a ação do veneno de Micrurus altirostris (cobra coral Uruguaiana) nas concentrações de 0,1; 0,5; 1; 5 e 10 µg/mL sobre a junção neuromuscular em preparações isoladas de ave (biventer cervicis de pintainho) e de mamífero (nervo frênico-diafragma de camundongo) e também a habilidade do antiveneno comercial produzido pelo Instituto Butantan e do anti-soro específico, produzido em coelhos em neutralizar a neurotoxicidade e a letalidade em pintainhos e camundongos (DL50 0,042 e 0,255 mg/kg), injetados i.m. e i.p., respectivamente, com misturas de veneno (5 DL50): antiveneno ou anti-soro específico (n=6), na razão de 1:1; 1:2,5; 1:5; 1:10 e 1:20. Somente a razão de 1:20 do veneno: antiveneno ou anti-soro neutralizou o bloqueio neuromuscular in vitro e protegeu pintainhos e camundongos contra 5 DL50 do veneno. O veneno causou um completo bloqueio neuromuscular em ambas as preparações, ligando-se aos receptores colinérgicos sub-sinápticos, de maneira irreversível. Inibiu as contraturas causadas por ação da acetilcolina e do carbacol, sem interferir nas respostas ao KCl. O bloqueio neuromuscular causado pelo veneno não apresentou qualquer alteração na ausência de íons Ca2+ ou com a diminuição da temperatura, tampouco foi revertido por ação da neostigmina. A resposta ao estímulo direto não foi afetada na vigência do bloqueio neuromuscular completo, indicando pela perfeita resposta muscular, que a membrana manteve-se íntegra, o que sugere a ausência de uma atividade miotóxica. Os resultados indicaram que o veneno de Micrurus altirostris interfere com a neurotransmissão pós-sináptica e que o antiveneno comercial, bem como o anti-soro específico, produzido em coelhos, possuem baixa eficácia em neutralizar a neurotoxicidade e a letalidade deste veneno. A neutralização da ação neurotóxica e também da letalidade do veneno foi obtida apenas quando a proporção de 1:20 (veneno:antiveneno) foi utilizada / Abstract: In this work, we studied the neuromuscular blockade caused by Micrurus altirostris venom (0.1-10 µg/mL) in indirect stimulated chick biventer cervicis and mouse phrenic nerve-diaphragm preparations and the ability of commercial antivenom (Instituto Butantan) and antiserum raised in rabbits to neutralize neurotoxicity and lethality in chicks and mice (LD50 0.042 and 0.255 mg/kg), injected i.m. and i.p., respectively, with venom (5 LD50):antivenom or antiserum mixtures (n=6) of 1:1, 1:2.5, 1:5, 1:10 and 1:20. The venom caused a complete and irreversible neuromuscular blockade in both preparations, binding in the subsynaptics colynergic receptors; inhibited the acetylcholine and carbachol contractures, without interfering on KCl response. The neuromuscular blockade was not Ca2+ or temperature-dependent and did not affect the response to direct stimulation. Only a venom:antivenom or antiserum ratio of 1:20 neutralized the neuromuscular blockade in vitro and protected chicks and mice against 5 LD50 of venom. Our results indicated that Micrurus altirostris venom interferes with postsynaptic neurotransmission and that commercial antivenom and rabbit antiserum have low efficacy in neutralizing the neurotoxicity and lethality of this venom / Doutorado / Doutor em Farmacologia

Histone deacetylases (HDACs) are key mediators of gene expression and, thus, major regulators of cell function. As such, HDACs play a role in orchestrating tumor biology, and the use of small inhibitors targeting theses proteins is attractive for the field of cancer therapy. Indeed, several HDAC inhibitors have received FDA-approval for the treatment of malignancies, while a myriad of these compounds continue to be evaluated in clinical trials. Besides their direct impact on tumor growth, HDAC inhibitors have been shown to increase immunogenicity of cancer cells, facilitating generation of a productive immune response against tumors. Immunotherapeutic approaches take advantage of the intrinsic ability of the immune system to manifest an anti-tumor response. Mechanisms of immune escape are often developed by cancer cells, neutralizing activity of the immune system. For example, upregulation of the PD1 ligands PDL1 and PDL2 by tumor cells negatively regulates the anti-tumor functions of PD1-expressing infiltrating T-cells. Importantly, strategies targeting this inhibitory axis have shown outstanding clinical benefit for the treatment of solid and hematological malignancies. The mechanisms by which HDAC inhibitors modulate tumor and immune cells biology were explored herein. Initially, treatment of melanoma cells with pan- and class I-selective HDAC inhibitors resulted in upregulation of PDL1 and PDL2 molecules. These effects were observed in mouse and human cell lines, as well as in tumor cells resected from metastatic melanoma patients. This upregulation was robust and sustained, lasting at least 96 hours in vitro, and validated in vivo using a B16F10 syngeneic mouse model. Enhanced expression of PDL1 mediated by HDAC inhibitors was found to result from enhanced histone acetylation at the PDL1 gene promoter region. Combination therapy of HDAC inhibition and PD1 blockade was explored in the tumor setting, leading to synergistic effects in terms of reducing melanoma progression and increasing survival of B16F10 melanoma-bearing mice. These data provide a clinical rationale for combination therapy of epigenetic modifiers (e.g. HDAC inhibitors) and PD1 blockade as means to augment cancer immunotherapy, improving patient outcomes. As a second pillar of this research, the impacts of HDAC-selective inhibition were explored on immune cell biology, since the broad nature of pan-HDAC inhibitors was shown to be detrimental to T-cells in vitro and in vivo. Based on screening assay results, novel implications of treating melanoma patient T-cells ex vivo with the HDAC6-selective inhibitor ACY1215 were investigated. Treatment with this compound was unique among pan- and isotype-selective HDAC inhibitors in modulating T-cell cytokine production and showing minimal impact of T-cell viability. ACY1215 tempered Th2 cytokine production (i.e. IL-4, IL-6 and IL-10), and maintained Th1 effector cytokines (e.g. IFNγ and IL-2). Furthermore, ACY1215 increased expression of surface markers, including CD69 activation marker and ICOS co-stimulatory molecule. In addition, ACY1215 treatment enhanced accumulation of central memory T-cells during ex vivo expansion of tumor infiltrating T-cells harvested from resected tumors of metastatic melanoma patients. Importantly, ACY1215-mediated inhibition improved tumor-killing capacity of T-cells. These results highlight an unexplored ability of selective HDAC inhibitor ACY1215 to augment T-cell expansion during protocols of adoptive cell therapy. While the discoveries presented here warrant further investigation of cellular and molecular mechanisms associated with ACY1215-treated T-cells, the clinic implications are clear and rapidly translatable.

Cette interrogation sur le blocus franco britannique porte sur l’efficacité du blocus, les mesures prises pour le mettre en oeuvre, sur sa réalité et sur sa régularité. Puis une fois ses approches traitées, on s’est intéressé à la mise en placedu blocus, aux tentatives de le forcer, à la collecte d’informations, à la diffusion de l’information, à la mise en place des réseaux d’informations. Ainsi à travers les décisions prises par l’Etat-major de la marine, on s’intéresse à la collecte del’information par les consulats, puis, relayée par les ambassades et les attachés navals, à l’information en elle-même pour savoir comment elle a contribué à la capture des navires ennemis. Différents aspects sont étudiés pour savoir comment lamarine allemande de commerce a réussi à adopter des stratégies pour contourner le blocus allié. Il convient aussi de s’intéresser aux neutres qui accueillent ces navires bloqués pour connaitre leurs regards sur des problèmes extérieurs. Les éléments spatio-temporels sont déterminants pour comprendre les dynamiques du blocus et les différentes politiques et moyens mis en oeuvre par les différents acteurs du conflit. Ainsi grâce à ces études, on tire un bilan des succès et des échecs de ce premier blocus naval à dimension mondiale en soulignant les éléments permettant sa réussite comme en soulignant ses failles / This questioning of the Anglo-French blockade applied to its effectiveness, the measures taken to keep it regularly. Once a study’s treated, This work is on the setting up of the blockade, on the attempts to cross it, on the datagathering, on thecirculation of the information, on the netting of a net information and on the efficiency of the blockade. Trough the decisions taken by the naval headquarters, I interested in the gathering of information coming from the consulate, serst to the embassies and the naval attaché. Her information by itself to know how it was leading to the capture of the enemy vessels. Various aspects are considered to know how the German Merchant Navy has succeeded to get around the allied blockade. It focuses also on Neutrals attitude toward the blockade, to know now their managed in front of this problem. The elements in time as well as in space are decisive for the comprehension of the blockade dynamics and the different politics and methods used by the actors of conflict. Theses study enables understand the success and this failures of the first world size naval blockade

Mes travaux de thèse portent sur la fabrication et la caractérisation électrique et magnétique de jonctions tunnel à base de graphène. C’est autour de l’idée d’apporter une meilleur compréhension des mécanismes d’injection et de détection d’un courant de charge et de spin aux interfaces graphène/ferromagnétique que s’articule ce manuscrit. Après avoir démontré qu’il est possible de faire croître de manière épitaxiée une barrière tunnel de MgO sur graphène, nous avons étudié les mécanismes de transport dépendant en spin dans des jonctions verticales de Co/MgO/Gr/Ni. Nous avons mis en évidence l’interaction du graphène avec l’électrode de nickel à travers les inversions de signe de la magnétorésistance. Celles-ci peuvent être expliquées à l’aide d’un modèle de canaux de conduction assistés par phonons. Enfin du blocage de Coulomb reproductible a été mesuré dans des amas d’aluminium potentiellement mono disperses et auto assemblés sur graphène. / My PhD thesis deals with the fabrication and the electric and magnetic characterizations of magnetic tunnel junctions based on graphene. The interaction of graphene with its close environment opens new possibilities for spintronics applications. The manuscript is focused on the improvement of the understanding of mechanisms involved in the injection and detection of a polarized spin current at the graphene/ferromagnetic interfaces. We show that it is possible to grow epitaxially MgO tunnel barrier on graphene. We study the spin transport mechanisms in vertical junctions of Co/MgO/Gr/Ni. The interaction of graphene with nickel electrode is probed through tunnel magnetoresistance inversions which can be explained by the activation of phonon assisted conduction channel. We also measure in vertical and lateral devices based on alumina barrier on graphene, reproducible Coulomb blockade processes linked to the presence of monodisperse aluminum clusters at the graphene edge.

Central mechanisms involving aldosterone - mineralocorticoid receptor (MR) activation mediate the increase in sympathetic tone after myocardial infarction (MI). We hypothesized that an increase in cardiac sympathetic activity (CSA) post MI facilitates cardiac sympathetic axonal sprouting, and that central MR blockade attenuates CSA and reduces cardiac sympathetic hyperinnervation post MI. Western blotting and qRT-PCR were used to assess protein and gene expression, and fluorescent immunohistochemistry was used to study changes in sympathetic innervation. Tyrosine hydroxylase (TH) and Norepinephrine transporter protein content in the non-infarcted base of the heart remained unaltered. In contrast, protein gene product (PGP 9.5) protein was significantly increased 2 fold in the base of the heart, and 6 fold in the peri-infarct area at 1 wk post MI, and associated with increased ubiquitin expression. Cardiac myocytes rather than sympathetic axons were identified as the main source of elevated PGP 9.5 expression. At the infarct border sympathetic hyperinnervation was observed with a 4 fold increase in growth associated protein 43 (GAP 43), a 2 fold increase in TH and a 50% increase in PGP 9.5 positive fibers when compared to the epicardial side of the left ventricle in sham rats. Central infusion of the MR blocker eplerenone at 5 ug/day for 9 days post MI markedly attenuated the increase in TH, GAP 43 and PGP 9.5 nerve densities at the infarct border. Central MR blockade may attenuate sympathetic hyperinnervation by several mechanisms, including decreasing CSA post MI, or affecting expression or function of nerve growth factor protein. Marked PGP 9.5 expression occurs in cardiomyocytes early post MI, which may contribute to the increase in ubiquitin and the early cardiac remodeling post MI.

Abstract In this thesis we study the interaction between radiation and matter using superconducting circuits that behave analogously with the conventional photon-atom interaction in quantum optics. The research is done with a system consisting of a waveguide resonator (radiation) strongly coupled to a transmon device (matter). We focus on the phenomena caused by strong coupling between the radiation and matter, and by driving the resonator to higher excited states with a strong monochromatic radiation. These have been studied little in the traditional radiation-matter systems. Increasing the strength of the monochromatic radiation drive, the dynamics of the system experiences a transition from the quantum to the classical regime. Also, the free-particle states of the transmon start being populated. In the weak driving limit, the transmon can be regarded as a two-state system. As a consequence, the resonator-transmon system is conventionally discussed in terms of the linear Jaynes–Cummings model. However, for strong coupling the Bloch–Siegert shift, caused by the terms neglected in the Jaynes–Cummings model, is strong and the Jaynes–Cummings model is insufficient for describing the dynamics of the system. We study the effects caused by strong coupling and the excitation of the higher transmon states instigated by the driving of the resonator. With reflection spectroscopy, we measure the absorption spectrum of the system and compare this with the spectrum calculated numerically using the Floquet–Born–Markov approach. We find that, in the region of the quantum-to-classical transition, the two-state approximation for the transmon is insufficient and the higher transmon states are necessary for accurate simulations. By calculating the average resonator occupation, we compare different numerical models: the Lindblad master equation, the Floquet–Born–Markov, and the semiclassical model. Coupling a transmon to a resonator shifts the energy levels of the resonator. This shift in the energy levels prevents the higher resonator states from being populated if the system is weakly driven with a frequency that is near the resonance frequency of the resonator. We simulate this photon blockade numerically and show that the blockade is substantially different for the two-state and multistate transmon approximations. / Original papers Original papers are not included in the electronic version of the dissertation. Pietikäinen, I., Danilin, S., Kumar, K. S., Vepsäläinen, A., Golubev, D. S., Tuorila, J., & Paraoanu, G. S. (2017). Observation of the Bloch-Siegert shift in a driven quantum-to-classical transition. Physical Review B, 96(2). https://doi.org/10.1103/PhysRevB.96.020501 http://jultika.oulu.fi/Record/nbnfi-fe201803073899 Pietikäinen, I., Danilin, S., Kumar, K. S., Tuorila, J., & Paraoanu, G. S. (2018). Multilevel Effects in a Driven Generalized Rabi Model. Journal of Low Temperature Physics, 191(5–6), 354–364. https://doi.org/10.1007/s10909-018-1857-8 http://jultika.oulu.fi/Record/nbnfi-fe2018061325770 Pietikäinen, I., Tuorila, J., Golubev, D. S., & Paraoanu, G. S. (2019) Quantum-to-classical transition in the driven-dissipative Josephson pendulum coupled to a resonator, Manuscript. https://arxiv.org/abs/1901.05655

BACKGROUND: Soft tissue sarcomas (STSs) are a diverse group of cancers that originate from mesenchymal tissue and are estimated to result in 13,130 new cases and 5,350 deaths this year. These neoplasms are hard to detect, which results in physicians struggling to treat late-stage STSs with a limited number of ineffective treatments. Currently, surgical excision is the primary treatment with radiation therapy administered when possible. However, even with optimal margins, the rate of recurrence is high, and the overall survival is low. There is a desperate need for new, more effective treatments. Immune checkpoint blockade (ICB) has recently had widespread success in treating melanoma, and in recent trials, SARC028 and Alliance A091401, have shown demonstrated activity of ICBs in STS in the neoadjuvant setting. Two histological subtypes of STS showed more promise than others: dedifferentiated liposarcoma (DDLPS) and undifferentiated pleomorphic sarcoma (UPS). Another issue plaguing the field of STS is that there is no universal indicator of response. The percentage of hyalinization found within the tumor was recently identified as a better marker of response than radiographic imaging or percent viable tumor. METHODS: This study was an investigator-initiated, single-center, randomized, open-label, phase II study (NCT03307616), in which 23 patients with either DDLPS of the retroperitoneum (RP) or UPS of the extremity/trunk (ET) were separated by disease into Cohort 1 and 2, respectively. Subjects in each cohort were randomized into two neoadjuvant treatment arms per cohort. Arm A (n=7) of Cohort 1 received nivolumab (anti-PD-1) monotherapy, while Arm B (n=7) of Cohort 1 received nivolumab and ipilimumab (anti-CTLA-4) combination therapy. Arm C (n=5) of Cohort 2 received nivolumab monotherapy and radiation therapy, whereas Arm D (n=4) of Cohort 2 received nivolumab/ipilimumab combination therapy and radiation therapy. A tumor biopsy was obtained before treatment, and another sample was taken during the primary treatment of surgical excision. These samples were processed and analyzed by a pathologist who determined the percentage of viable tumor, hyalinization, and necrosis in each sample. Radiographic imaging was also taken throughout to make RECIST 1.1 response determinations. RESULTS: The average treatment response (1 - % viable tumor) for Cohort 1 was 25 ± 23 and there was no difference between Arm A and Arm B, p=0.48. The average treatment response in Cohort 2 was higher at 85 ± 27, but there was also no significant difference between the arms, p = 0.46. The mean percent hyalinization for Cohort 1 was 13 ± 13%, and for Cohort 2 was 69 ± 35%. Again, there was no significant difference between the arms in the Cohort 1 or 2, p = 0.45 and p = 0.54, respectively. Lastly, the mean % necrosis in Cohort 1 was 13 ± 13 %, and in Cohort 2 was 17 ± 24%, and neither had significantly different results in the arms, p = 0.60 and p = 0.92. The RECIST 1.1 results were independent of the arms of the study, and the radiographic response (percent image change) did not correlate to any metric of histologic response. Those who received Ipilimumab had higher rates of adverse events. CONCLUSION: There is significant evidence that ICBs elicited a response in RP DDLPS and ET UPS, and the response of ET UPS was profound. However, there was no apparent benefit from the combination therapy compared to the monotherapy in either cohort. The higher response in ET UPS may be due to the additional radiation therapy or to the nature of UPS itself. Finally, radiographic imaging does not show the response which is apparent at the histological level, so treatment regimens and future experiments should no longer rely on radiographic imaging as a marker for response. / 2021-06-07T00:00:00Z

Introduction The joint dysfunction of the spine in the sense of functional blockade is connected with corresponding painful condition, reflex contraction of the surrounding muscles and restriction of mobility in the spinal segment. Joint block has its mechanical context, which need not be accompanied by a structural disturbance but it's affecting the biomechanical properties of the spine. Several theories exist how the joints become restricted. Scientific evidence highly supports the meniscoid entrapment theory. Purposes The aim of the study was to identify the meniscoids of the cervical spine using in- vivo MRI imaging and to determine their potential role in the development of functional joint blocks of the axial system (AS). Another objective was to find out how the articular blocks affect the rheological properties of the spine by the Transfer Vibration through the Spine (TVS) method. Patient sample The MRI study was conducted on a research file of 12 subjects - two anatomical preparations and ten in vivo. Two subjects were examined in TVS experiment. Methodology Two anatomical preparations were investigated on MRI to find the appropriate sequence for imaging of meniscoids, three subjects for identification of meniscoids in vivo. Seven subjects underwent initial investigation, manipulation of...

This diploma theses attempts to find answers to the question: How does the media represent environmental activists? Using the anti-logging protests in Šumava National Park that resulted in blockades in 1999 and 2011 as an example. The author tries to capture the background of the affair, and the view of Czech society towards environmental activists, which is significantly influenced by the media. The analytical part combines qualitative and quantitative research methods based on content analysis of two selected daily news sources (Mladá fronta Dnes and Právo). Emphasis is given to the dynamics of the affair, which is why longer time periods are analyzed, comprising one year before and two years after each blockade. Key words: Šumava, media, environmental activists, blockade, content analysis