Negatively biased facial affect discernment may prompt socially inhibited behavior. Characterizing normative patterns of facial affect discernment across emotions and expression intensity during middle childhood will help to identify subtle, yet meaningful, deviations that may emerge for individuals and potentially negatively impact their social behavior. Facial affect discernment for happy, sad, and angry expressions across low, medium, and high intensities and parent-reported socially inhibited behavior were measured in this study in a sample of 7-10 year-old children (N = 80; 53% female). Discernment accuracy improved with increased expression intensity for all emotions. Specifically, we found a quartic effect for the association between intensity and accuracy for anger and negative quadratics effects with decelerating positive rates of changes for associations between intensity and accuracy for happiness and intensity and accuracy for sadness. Additionally, discernment accuracy for happiness was generally better than for sadness and anger; discernment accuracy for anger was generally better than for sadness. However, at low intensity, discernment accuracy for sadness was comparable to accuracy for happiness but better than for anger. Neither misidentification of neutral and low intensity faces as negative nor discernment accuracy of happiness at low intensity was significantly associated with socially inhibited behaviors. Although accurate discernment of anger and sadness at low intensity was not significantly related to socially inhibited behavior, better discernment accuracy of anger and sadness at medium intensity was significantly related to more socially inhibited behavior. Overall, these results enhance understanding of normative facial affect discernment and its relation to maladaptive social behaviors in middle childhood, a developmental stage at which intervention efforts may prove effective at heading off detrimental outcomes associated with socially inhibited behavior such as loneliness, low self-esteem, peer victimization, social anxiety, and depression that increase in late childhood and adolescence.

The response of frog sciatic nerve action potentials to x-irradiation and metabolic (carbon monoxide) or transport (ouabain) inhibition was determined in an attempt to further identify the nature of radiation insult to nervous tissue. Carbon monoxide, ouabain (2 X 10-5 M), and nitrogen anoxia were shown to produce a near linear decline in action potential amplitude. The carbon monoxide and nitrogen inhibitions of activity were reversible in air; the carbon monoxide inhibition was light reversible. Ouabain inhibition was partially reversible by soaking the nerve in aerated Ringer's. Application of 120 kv x-rays (75 Kr at 4.9 Kr/min) to nerves during the linear decline in spike amplitude brought about a marked enhancement (146%) of inhibition by 99% CO/l% 02, nitrogen (136%), and ouabain (265%). All bhanges were shown to be statistically significant by a regression analysis. However, x-irradiation did not appear to alter the air reversibility of carbon monoxide and nitrogen inhibitions nor the reversibility in Ringerts of the ouabain inhibition. Additionally x-irradiation completely blocked light reversal of 98% CO/2% 02 inhibition and produced a decline in activity. A possible interpretation of these results is a compensation for radiation action at this dosage requiring metabolism and ion pump activity.

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

Radiotherapy continues to be a primary modality in the treatment of cancer. DNA damage induced by radiation can promote apoptosis as well as both autophagy and senescence, where autophagy and senescence can theoretically function to prolong tumor survival. A primary aim of this work was to investigate the hypothesis that autophagy and/or senescence could be permissive for DNA repair, thereby facilitating tumor cell recovery from radiation-induced growth arrest and/or cell death. In addition, studies were designed to elucidate the involvement of autophagy and senescence in radiation sensitization by PARP inhibitors and the re-emergence of a proliferating tumor cell population. In the context of this work, the relationship between radiation-induced autophagy and senescence was also determined. Studies were performed using DNA repair proficient HCT116 colon carcinoma cells and a repair deficient Ligase IV (-/-) isogenic cell line. Irradiation promoted a parallel induction of autophagy and senescence that was strongly correlated with the extent of persistent H2AX phosphorylation in both cell lines; however inhibition of autophagy failed to suppress senescence, indicating that the two responses were dissociable. Irradiation resulted in a transient arrest in the HCT116 cells while arrest was prolonged in the Ligase IV (-/-) cells; however, both cell lines ultimately recovered proliferative function, which may reflect maintenance of DNA repair capacity. The PARP inhibitors (Olaparib) and (Niraparib) increased the extent of persistent DNA damage induced by radiation as well as the extent of both autophagy and senescence; neither cell line underwent significant apoptosis by radiation alone or in the presence of the PARP inhibitors. Inhibition of autophagy failed to attenuate radiation sensitization, indicating that autophagy was not involved in the action of the PARP inhibitors. As with radiation alone, despite sensitization by PARP inhibition, proliferative recovery was evident within a period of 10-20 days. While inhibition of DNA repair via PARP inhibition may initially sensitize tumor cells to radiation via the promotion of senescence, this strategy does not appear to interfere with proliferative recovery, which could ultimately contribute to disease recurrence.

Le bulbe olfactif est principalement constitué de neurones excitateurs, dits cellules mitrales, interconnectés via des inter-neurones inhibiteurs, dits cellules granulaires. L'analyse de données expérimentales recueillies en condition in vitro dans des tranches de bulbe olfactif de rats révèle que le caractère phasé des potentiels d'action des cellules mitrales relativement aux oscillations du potentiel de champ local est largement atténué lorsque l'on bloque pharmacologiquement l'inhibition provenant des granules, mettant ainsi en évidence le rôle primordial de l'inhibition synaptique. Les fluctuations de la conductance synaptique inhibitrice sont corrélées à celles mesurées sur le potentiel de champ local. Une relation entre l'inhibition reçue et la phase des potentiels d'action est dévoilée. Un neurone aura plus de chance d'émettre en phase s'il reçoit un nombre important d'événements synaptiques inhibiteurs et si ces événements sont eux-même phasés. Ces résultats sont rassemblés au sein d'un modèle informatique de bulbe olfactif afin d'explorer les capacités de codage de l'interaction mitrale-granule. Après avoir montré que le transfert d'information des cellules mitrales semble plus résider dans leurs instants précis d'émission de potentiels d'action au cours des oscillations que dans leurs fréquences de décharges, une étude analytique conclut que la robustesse du code produit par les cellules mitrales lors des oscillations du réseau est conditionnée par une forte interaction synaptique. Nous appliquons notre modèle de bulbe olfactif pour reconnaître des odeurs à l'aide d'une matrice de capteurs de gaz artificiels. / The olfactory bulb is mainly composed of excitatory cells, called mitral cells, interconnected via local inhibitory neurons, called granule cells. The analysis of electrophysiological data, recorded in vitro from rat olfactory bulb slices, shows that mitral cell firing is phase-locked to the fast local field potential oscillation. This phase-locking is largely reduced when the inhibitory synaptic conductance is pharmacologically blocked, hence highlighting the important role of synaptic inhibition. We find that the inhibitory conductance fluctuations are correlated to the local field potential oscillations. A relationship between the received inhibition and the phase of mitral action potentials is also revealed. The probability to fire a phase-locked action potential increases if the neuron receives a large number of inhibitory synaptic events, and if these events are themselves phase-locked. Results from the previous analysis are used to design a computational model of the olfactory bulb in order to explore the encoding capacity of the mitral-granule interplay. It appears likely that mitral cells encode information in precise spike timings rather than in firing rates. We therefore study analytically the influence of the number and the temporal dispersion of the received inhibitory synaptic events on the spike timing precision of mitral cells. Our study concludes that spike timing precision requires a strong synaptic coupling between mitral and granule cells. Lastly, our olfactory bulb model is applied to the recognition of odours by using an array of artificial gas sensors.

UNIVERZITA KARLOVA V PRAZE FAKULTA TĚLESNÉ VÝCHOVY A SPORTU Objektivizace svalového napětí při použití kinesiotapingu pomocí myotonometru Diplomová práce Vedoucí diplomové práce: Vypracovala: PhDr. Petr Šifta, PhD. Bc. Kristýna Fleišmanová Praha, duben 2012 2 Prohlašuji, že jsem diplomovou práci zpracovala samostatně a že jsem uvedla všechny použité informační zdroje a literaturu. Tato práce ani její podstatná část nebyla předložena k získání jiného nebo stejného akademického titulu. V Praze, dne …………………… …..…………………… Bc. Kristýna Fleišmanová 3 EVIDENČNÍ LIST KNIHOVNY Souhlasím se zapůjčením své diplomové práce ke studijním účelům. Uživatel svým podpisem stvrzuje, že tuto diplomovou práci použil ke studiu a prohlašuje, že ji uvede mezi použitými prameny. Jméno a příjmení: Fakulta / katedra: Datum vypůjčení: Podpis: 4 Děkuji PhDr. Petru Šiftovi, PhD. za cenné rady, připomínky a podněty, které mi pomohly orientovat se v dané problematice, a tak ulehčily a zkvalitnily zpracování této diplomové práce. 5 ABSTRAKT Název: Objektivizace svalového napětí při použití kinesiotapingu pomocí myotonometru Cíle práce: Cílem této diplomové práce je shrnout teoretické poznatky týkající se dané problematiky a vypracovat pilotní studii zaměřenou na ověření facilitační a inhibiční funkce v kinesiotapingu. Experiment má na...

The inhibitor of apoptosis proteins (IAPs) have been shown to interact with a growing number of intracellular proteins and signalling pathways in order to fulfil their anti-apoptotic role. In order to investigate in detail how the avian homologue ITA interfered with both TNF induced apoptosis and the NGF mediated differentiation in PC12 cells, a two hybrid screen was performed with a PC12 library using ITA as a bait. The screen resulted in the identification of several overlapping fragments of a previously unknown gene. The complete cDNA for this gene was isolated, the analysis of which revealed a high homology with a large family of tumour antigens known as MAGE (melanoma associated antigens). This newly identified member of the MAGE family, which was later named NRAGE, exhibited some unique characteristics that suggested for the first time a role in normal cellular physiology for this protein family. MAGE proteins are usually restricted in their expression to malignant or tumour cells, however NRAGE was also expressed in terminally differentiated adult tissue. NRAGE also interacted with the human XIAP in direct two-hybrid tests. The interactions observed in yeast cells were confirmed in mammalian cell culture, employing both coimmunoprecipitation and mammalian two-hybrid methods. Moreover, the results of the coimmunoprecipitation experiments indicated that this interaction requires the RING domain. The widely studied 32D cell system was chosen to investigate the effect of NRAGE on apoptosis. NRAGE was stably transduced in 32D cells, and found to augment cell death induced by the withdrawal of Interleukin-3. One reason for this reduced cell viability in NRAGE expressing cells could be the binding of endogenous XIAP, which occurred inducibly after growth factor withdrawal. Interestingly, NRAGE was able to overcome the protection afforded to 32D cells by the exogenous expression of human Bcl-2. Thus NRAGE was identified during this research doctorate as a novel pro-apoptotic, IAP-interacting protein, able to accelerate apoptosis in a pathway independent of Bcl-2 cell protection. / Die Familie der „inhibitor of apoptosis proteins” (IAPs) interagieren mit einer wachsenden Zahl an intrazellulären Proteinen und Signaltransduktionswegen um ihre anti-apoptotische Aufgabe zu erfüllen. Es konnte gezeigt werden, dass das ITA-Homolog aus dem Huhn sowohl die durch TNF induzierte Apoptose als auch die durch NGF induzierte Differenzierung von PC12-Zellen verhindert bzw. verlangsamt. Um diese Befunde genauer zu untersuchen, wurde in dieser Arbeit ein "yeast two-hybrid screen" mit ITA als "bait" (Köder) und einer cDNA-Bank aus PC12-Zellen durchgeführt. In diesem "screen" konnten verschiedene Fragmente eines bis dahin unbekannten Gens identifiziert werden. Die Untersuchung der isolierte Gesamt-cDNA ergab eine hohe Homologie mit einer Familie von Tumor-Antigenen namens MAGE (melanoma associated antigens). Im Gegensatz zu den bisher identifizierten MAGE, zeigte dieses neue Familienmitglied, welches später NRAGE genannt wurde, einige Charakteristika die das erste mal eine Rolle in der normalen zellulären Physiologie nahe legten. In einem direkten "yeast two hybrid test" konnte die Interaktion zwischen NRAGE und humanem XIAP gezeigt werden. Diese Interaktion konnte sowohl in Ko-Immunpräzipitationen als auch im sogenannten Säuger two-hybrid bestätigt werden. Desweiteren zeigten die Ko- Immunpräzipitationen, dass für die Interaktion dieser beiden Proteine die RING-Domäne von ITA benötigt wird. Um Effekte von NRAGE auf die Apotose zu untersuchen, wurde dass etablierte 32D Zellsystem verwendet. NRAGE wurde stabil in 32D-Zellen exprimiert, wodurch die Apoptoserate der Zellen, induziert durch die Kultivierung in IL-3 freiem Medium, gesteigert wurde. Ein Grund für diese erhöhte Apoptoserate, könnte in der Bindung von XIAP, welches nach dem Entzug von Wachstumsfaktoren induziert wird, an NRAGE liegen. Interessanterweise war NRAGE auch fähig die protektive Wirkung von exogen exprimiertem Bcl-2 aufzuheben. Somit konnte in dieser Arbeit NRAGE als pro-apoptotisches und mit IAP interagierendes Protein beschrieben werden. Desweiteren konnte gezeigt werden, dass die Verstärkung der Apoptose unabhängig von dem bekannten durch Bcl-2 bedingten anti-apoptotischen Signalweg erfolgt.

ABSTRACT The magnitude of the AIDS epidemic is well documented. It has been shown that Africa constitutes about 70 % of people infected with HIV worldwide. Efforts to control the AIDS epidemic have focused heavily on studies pertaining to the biology, biochemistry and structural biology of HIV and on the interactions between HIV proteins and new drugs. One of the most challenging problems in AIDS therapy is that HIV develops drug-resistant variants rapidly. Extensive research has been dedicated to designing resistance-evading drugs for HIV-1 protease (predominantly subtype B), which is crucial for the maturation of viral, structural and enzymatic proteins. There are 10 subtypes of HIV-1 within the major group of the virus, with subtype C accounting for about 95 % of infections in South Africa. Since HIV-1 antiretroviral treatment has been developed and tested against the B subtype, which is prevalent in North America, Western Europe and Australia, an important question relates to the effectiveness of these drugs against the C subtype. At this point, however, little is known about inhibitor-resistant mutations in the subtype C. The study, therefore, looked at the two active site mutations (V82A and V82F/I84V) in the South African HIV-1 subtype C protease (C-SA) emerging from the viral population circulating in patients. These mutations are well-characterized within the framework of the subtype B and are known to cause cross-resistance to most of inhibitors currently in clinical use. Protein engineering techniques were used to generate the V82A and the V82F/I84V variants. Comparative studies with the wild-type HIV-1 C-SA protease were performed. The spectral properties of the V82A and the V82F/I84V variants indicated no changes in the secondary structure in the respective variant proteins. Tryptophan and tyrosine fluorescence indicated a major difference in the intensities at the emission maxima for all three proteins. The fluorescence intensity of the V82F/I84V variant, in particular, was significantly enhanced indicating the occurrence of tertiary structural changes at/near the flap region. Both mutations did not impact significantly upon catalytic function. Both variants also had the same Km values comparable to that of the wild-type enzyme. The catalytic efficiencies and the kinetic constants were lowered 3.6-fold for the V82A mutation and 6-fold for the V82F/I84V mutation relative to the wild-type C-SA protease. Inhibition studies were performed using four inhibitors in clinical use (saquinavir, ritonavir, indinavir and nelfinavir). For the V82A variant, IC50 and Ki values for saquinavir and nelfinavir iv were not affected, whilst those for ritonavir and indinavir were 5- and 9-fold higher than the wild-type C-SA protease, respectively. Against the V82F/I84V variant, however, the inhibition constants were drastically weaker and characterized by IC50 and Ki ratios ranging from 50 to 450. Isothermal titration calorimetry (ITC) was also used to determine the binding energetics of saquinavir, ritonavir, indinavir and nelfinavir to the wild-type C-SA, V82A and V82F/I84V HIV-1 protease. The V82A mutation lowered the Gibbs energy of binding for the respective four clinical inhibitors by 0.4 kcal/mol, 1.3 kcal/mol, 1.5 kcal/mol and 0.6 kcal/mol, respectively, relative to the wild-type C-SA HIV-1 protease. The affinity of V82A HIV-1 protease for saquinavir, ritonavir, indinavir and nelfinavir (Kd = 1.85 nM, 2.00 nM, 12.70 nM and 0.66 nM, respectively, at 25 °C) was in the range of 2- to 13-fold of magnitude weaker than that of the wild-type C-SA protein. The clinical inhibitors exhibited the highest binding affinity to both the wild-type and the V82A enzymes, but were extremely sensitive to the V82F/I84V mutation. The V82F/I84V mutant reduced the binding of saquinavir, ritonavir, indinavir and nelfinavir 117-, 1095-, 474- and 367- fold, respectively. A drop in Kd values obtained for the V82F/I84V in association with saquinavir, ritonavir, indinavir and nelfinavir was consistent with a decrease of between 2.8 - 4.2 kcal/mol in ΔG, which is equivalent to at least 2 to 3 orders of magnitude in binding affinity. Taken together, thermodynamic data indicated that the V82A and V82F/I84V active site mutations in the C-SA subtype lower the affinity of the first-generation inhibitors by making the binding entropy less positive (unfavorable) and making the enthalpy change slightly less favorable.

A procura por suplementos alimentares cresce a cada ano, pois o seu uso contínuo promove a melhora e manutenção da qualidade de vida do hospedeiro. Sustentado neste princípio, a ingestão de produtos probióticos, principalmente leites fermentados, passa a ser uma alternativa neste segmento do mercado. Os probióticos são suplementos alimentares que contem microrganismos vivos que trazem benefícios à saúde do consumidor pela manutenção e melhora do balanço microbiano do trato gastrintestinal. Existem vários gêneros de microrganismos que apresentam propriedades probióticas, destacando-se o gênero Lactobacillus; que exercem várias funções benéficas ao hospedeiro como a diminuição dos níveis de colesterol sangüíneo, efeito anticarcinogênico, adesão ao epitélio intestinal, melhora do sistema imune e a exclusão competitiva, conhecida também como efeito barreira. Dentro deste contexto, o presente trabalho teve como objetivo avaliar o efeito de inibição exercido por cinco cepas de Lactobacillus isoladas de fezes humanas: denominadas L. plantrum (Lac-01), L. plantarum (Lac-02), L. fermentum (Lac-03), L. fermentum (Lac-04) e L. fermentum (Lac-05) sobre o crescimento de E. coli O157:H7, Listeria spp., P. aeruginosa ATCC 27853, Salmonella typhi e Shigella spp , por meio da técnica de co-cultura. Os resultados revelaram que as cepas Lac-01, Lac-02 e Lac-03 foram capazes de inibir, em diferentes níveis, o crescimento das cepas patogênicas avaliadas, sendo este efeito mais acentuado para S. typhi e Shigella spp. Observou-se ainda que a cepa Lac-04 não apresentou efeito de inibição sobre E. coli O157:H7 e P. aeruginosa, ao passo que a cepa Lac-05 inibiu o crescimento de P. aeruginosa, S. typhi e Shigella spp., sendo incapaz de inibir E. coli O157:H7 e Listeria spp, nas condições estudadas. Posteriormente ao se verificar a produção de substâncias antimicrobianas por meio do método \"spot-test\", observou-se o efeito positivo de inibição exercida pelas cepas de Lactobacillus sobre o crescimento dos respectivos patógenos. Verificou-se também, por meio de testes enzimáticos específicos, que as substâncias antimicrobianas produzidas pelas cepas de Lactobacillus não foram sensíveis às enzimas utilizadas, não podendo afirmar, desta forma, que estas substâncias são bacteriocinas. Os testes de co-agregação demonstraram que as cepas Lac-04 e Lac-05 exerceram melhor efeito sobre os patógenos, quando comparadas com as cepas Lac-01 e Lac-02. A cepa Lac-03 não exerceu efeito de coagregação com nenhum dos patógenos avaliados. Os testes de autoagregação revelaram que as cepas Lac-01, Lac-02, Lac-04 e Lac-05 exerceram efeito de autoagregação acentuado. Observou-se ainda que nenhum dos patógenos estudados foi capaz de autoagregar. / The probiotics are special food that contain live microorganisms that promote beneficits to consumer health through maintenance and improvement of microbial balance of gut tract. There are many microorganisms species used in probiotics products, standing out Lactobacillus species that produce many beneficies to consumer like decrease of blood cholesterol level, anti-carcinogenic effect, adhrence to intestinal epithelium, immune system stimulation and the competitive exclusion, also known as barrier effect. In this regard, the present work aimed to evaluate the inhibition effect exerted by five Lactobacillus strains, isolated from humam faeces: denominate, L. plantarum Lac-01, L. plantarum Lac-02, L. fermentum Lac-03, L. fermentum Lac-04 e L. fermentum Lac-05 on the gowth of E. coli O157:H7, Listeria spp., Pseudomonas aeruginosa ATCC 27853, Salmonella typhi and Shigella spp., by means of coculture technique. The results showed that the strains Lac-01, Lac-02 and Lac-03, were able to inhibit, at different levels, the growth of pathogenic strains, and this effect was more accentuated on S. typhi and Shigella spp. It was also observed that the strain Lac-04 did not present inhibition effect on E. coli O157:H7 and P. aeruginosa ATCC 27853, and the Lac-05 inhibited the growth of P. aeruginosa, S. tyhpi and Shigella spp., and showed no effect on E. coli O157:H7 and Listeria spp. Afterwards, the production of antimicrobial substance was verified by means of \"spot-test\" method, showing the positive inhibition effect exerted by all Lactobacillus strains on the growth of the respective pathogens. By means of specific enzymatic test, the antimicrobial substances produced by lactobacillus strains were not sensible to the action of enzymes proteinase-K, papain and pepsin, but this is not enough to affirm that this substances are bacteriocins. The coaggregation test demonstrated that the strains Lac-04 and Lac-05 exerted better effect on the pathogens, when compared to strains Lac-01 and Lac-02. The strain Lac-03 did not coaggregate with any pathogenic strains. The autoaggregation test showed that the strain Lac-01, Lac-02, Lac-04 and Lac-05 exerted accentuated auto-aggregation effect. The studied pathogen was not able to autoaggregate.

The nature and assessment of executive function are areas of active research. Many current assessments of executive function are complex, have limited reliability and validity, and suffer from task impurity, meaning other cognitive processes may indirectly influence task performance. Additionally, measures may be culture, language, or education bound limiting their use in certain populations (Miyake, Emerson, & Friedman, 2000; Miyake, Friedman, et al., 2000; Strauss, Sherman, & Spreen, 2006; Stuss, 2007). The purpose of this project was to develop a novel set of executive function measures to address issues with current clinical measures. The new measures 1) can be used in an ERP environment, 2) use the same stimulus set to address task impurity and 3) use simpler cognitive operations of inhibition, set-shifting, and updating, identified in previous research by Miyake et al., (2000). Twenty-nine undergraduate participants at the University of South Florida were administered currently used clinical measures of executive function theorized to engage in inhibition, set-shifting, and updating and the set of the novel tasks. ERP data was collected during the administration of the novel tasks. Behaviorally, conditions theorized to engage executive function resulted in slower response reaction time than control conditions. Additionally, behavioral results indicated that performance on novel tasks were differentially related to different clinical EF tasks. ERP differences were observed between both Go/No-Go conditions (inhibition) and among N-back conditions (updating). Results suggest the novel executive function tasks are tapping into different cognitive processes and may be a viable tool for studying executive function in the future.