Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / CHAPTER 2: This study examined the variation in the cortisol awakening response (CAR) in relation to mood states and performance during professional swimming competition. Eleven athletes were examined during two consecutive days of competition. On each day salivary cortisol was determined upon awakening, 30 and 60 min post awakening, immediately before warming up for competition and 5, 20 and 60 min after competition. Psychometric instruments included the Profile of Mood States and self-reports of performance. CARs did not differ between competition and non-competition days and were not related to performance on any day. However, difference was observed in the concentration of cortisol prior to and after the contest between competition and non-competition days. Perceived demands of the day ahead might not reflect the same magnitude of variation in the CAR in well-trained men. Explanations for this include better coping mechanisms and response towards the phase and time of competition. CHAPTER 3: Objective: We investigated the response of salivary total protein (TP), alpha-amylase (sAA) and chromogranin A (CgA) to sporting competition and their relation with positive and negative affect. Methods: Eleven professional swimmers were examined during the first day of a national contest and on a recreated event that matched time-of-the-day and day-of-theweek assessments two weeks later. Total protein was determined by the Bradford method and sAA and CgA by western blotting upon awakening, 30 and 60 min post awakening, immediately before warming up for competition and 5, 20 and 60 min after competition. Psychometric instruments included the Positive Affect and Negative Affect Schedule - X (PANAS-X). Results: The concentrations of TP, sAA and CgA differed from controls only prior to and 5 min after the event. We observed associations between higher negative affect scores with higher levels of TP, sAA and CgA prior to the event on the competition day. Areas under the curve did not differ from controls for TP, sAA or CgA. Conclusion: TP and CgA showed a similar reactivity to sporting competition than sAA, which may be attributed to the mechanisms responsible for protein secretion into saliva when collection is performed with no exogenous stimulation. Strong adverse psychological stimuli only seem to override the regular rhythm of salivary proteins moments before and after stressful situations. / CAPÍTULO 2: O presente estudo investigou a variação da resposta do cortisol ao acordar (CAR) em relação com estados de humor e desempenho durante uma competição de natação profissional. Onze atletas foram examinados durantes dois dias consecutivos de competição. Em cada dia, a concentração de cortisol salivar foi determinada ao acordar, 30 e 60 minutos após acordar, imediatamente antes de aquecer para a competição e 5, 20 e 40 minutos depois da competição. O Perfil de Estados de Humor e reportes próprios de desempenho foram incluídos como instrumentos psicométricos. A CAR não foi diferente entre os dias de competição e controle e não houve relação com o desempenho em nenhum dos dois dias. No entanto, houve diferença antes de e após o evento entre os dias de competição e controle. A percepção de um desafio que irá acontecer durante o dia não necessariamente reflete a mesma magnitude de variação na CAR em sujeitos bem treinados. Isto pode ser devido a melhores mecanismos para lidar com o estresse e a fase e hora da competição em que os sujeitos foram avaliados. CAPÍTULO 3: O presente estudo investigou a resposta de proteína total na saliva (TP) alfaamilase (sAA) e cromogranina A (CgA) à competição esportiva e sua relação com estados de humor. Onze atletas profissionais foram examinados durante o primeiro dia de um evento nacional em natação e durante um evento simulado num dia de treinamento no mesmo dia da semana e no mesmo horário que a competição real. A concentração de TP foi determinada pelo método de Bradford e a de sAA e CgA por western blotting ao acordar, 30 e 60 minutos depois, imediatamente antes de aquecer para o evento e 5, 20 e 40 minutos depois do mesmo. A escala de estado de humor PANAS-X (Positive Affect and Negative Affect Schedule) foi incluída como instrumento psicométrico. A concentração de TP, sAA e CgA foi diferente dos dias de controle unicamente antes e 5 minutos após a competição. Não houve diferença na área sob a curva do perfil diurno de cada marcador entre os dias de competição e controle. TP e CgA tiveram uma resposta similar à competição do que sAA. Isto pode ser atribuído aos mecanismos de secreção de proteína na saliva quando a coleta da mesma é feita sem estimulação exógena. Estímulos psicológicos adversos parecem alterar o ritmo regular de secreção destes marcadores só momentos prévios e posteriores a situações estressantes. / Mestre em Genética e Bioquímica

Tick-borne encephalitis is a serious disease of the central nervous system. It is caused by tick-borne encephalitis virus, which is transmitted by ticks. The Czech Republic is one of the countries with the highest prevalence of this disease. Tick-borne encephalitis virus is able to replicate in several cell types. In this work we focused on macrophage line IC-21 and dendritic cells, because these cells are the first, which encounter the virus and support its spreading in the host at early stage of infection. So far there is not known any specific receptor for virus entry into cells or which signaling pathways activates. Therefore, we decided to investigate the activation of selected signaling pathways after infection with tick-borne encephalitis virus and influence of tick saliva on this activation. We employed methods of dual luciferase reporter assay, immunosandwich assay and western blot. The obtained results showed that in virus infected IC-21 cells are activated phosphatidyl-inositol pathway, NF-κB pathway, signaling molecule Erk1/2 and others. Testing of tick saliva effect revealed significantly decreased activity of NF-κB, AP-1 and CREB.

We often form intentions but have to postpone them until the appropriate situation for retrieval and execution has come, an ability also referred to as event-based prospective memory. After intention completion, our cognitive system has to deactivate no-more-relevant intention representations from memory to avoid interference with subsequent tasks. In everyday life, we frequently rely on these abilities also in stressful situations. Surprisingly, little is known about potential stress effects on these functions. Therefore, the present study aimed to examine the reliability of event-based prospective memory and of intention deactivation in conditions of acute psychosocial stress. To this aim, eighty-two participants underwent the Trier Social Stress Test, a standardized stress protocol, or a standardized control situation. Following this treatment, participants performed a computerized event-based prospective memory task with non-salient and focal prospective memory cues in order to assess prospective memory performance and deactivation of completed intentions. Although the stress group showed elevated levels of salivary cortisol as marker of a stress-related increase in hypothalamus-pituitary-adrenal axis activity throughout the cognitive testing period compared to the no-stress group, prospective memory performance and deactivation of completed intentions did not differ between groups. Findings indicate that cognitive control processes subserving intention retrieval and deactivation after completion may be mostly preserved even under conditions of acute stress.

16-androstenes Anosmia in Men 16-androstens are compounds of steroid character which are secreted to the skin as part of apocrine glands secrets and they participate in creating body odor. It was found that androstenes influence affective state and psychophysiological reactions. The effect of androstenes can be also found on subliminal level. However, it has not yet been tested whether individuals with specific anosmia could also be influenced this way. Specific anosmia is described as an individual's incapability of conscious detection of particular odor while the detection of other compounds remains unaffected. The aim of the thesis is to test whether specifically anosmic individuals can be influenced by compounds to which they are anosmic to. This hypothesis is based on research of sensitization. Previous research indicates that olfactory perception may be of larger plasticity than thought. The theoretical part focuses on critical analysis of androstenes and research on specific anosmia. This field contains topics such as influence of chemical structure on androstenes perception, genetic influences, sensitization, functions of androstenes in interpersonal interactions and research on brain imagining. The experimental part focused on measuring subjective ratings of mood and psychophysiological...

No description available.

No description available.

Indiana University-Purdue University Indianapolis (IUPUI) / Background: Bacterial adherence to the acquired dental pellicle, important in caries, is mediated by receptor-adhesin interactions such as Streptococcus mutans antigen I/II (I/II). Ten I/II epitopes from the A, V, P and C regions were chosen to determine their reactivity in human saliva. Underlying the body’s ability to immunologically respond to bacteria that lead to caries are the human leukocyte antigen (HLA) genes, specifically HLA class II (HLA-II) genes that control antigen presentation. Previous studies suggested that a specific HLA biomarker group (HLA-DRB1*04) may have differential control of immune responses to I/II. However, it was not known whether secretory IgA (SIgA) responses to the selected epitopes from HLA-DRB1*04 positive subjects were different compared to their non-biomarker counterparts (negative), or across other caries factors, since no study to date had thus assessed these questions. Methods: Per IRB approval, the study population was divided into age, sex and race matched DRB1*04 positive (n=16) and negative groups (n=16). SIgA-epitope (and whole cell) reactivity was determined using ELISA. Other caries factors were measured. Subjects received a clinical exam by a trained examiner. ix Differences between DRB1*04 positive and negative groups were examined using a two-sided, two-sample t-test. Results: DRB1*04 positive subjects had numerically, but not statistically, higher reactivity to 9 out of 10 epitopes, the exception being residues 834-853 from the V and P regions of I/II across multiple measures. Though statistically insignificant, DRB1*04 positive subjects also exhibited 25-30 μg mL-1 less total IgA (TIgA) than negative counterparts. All clinical caries data proved inconclusive when comparing groups, likely due to exogenous factors and sample size. Conclusion: DRB1*04 positive subjects showed a trend toward lower TIgA. Moreover, they also showed a lower SIgA response across multiple measures to 834-853, the I/II V and P region epitope. This region forms a sort of functional epicenter involved in collaboration between domains along the entire I/II antigen, and governs the region involved in initial attachment to the acquired dental pellicle. This region may be involved in an in vivo discontinuous conformationally specific immunogenic epitope that serves as an HLA-II binding motif which remains elusive.

Democratic and accountable policing is one of the hallmarks of democracy. In a healthy democracy, a police service exists to protect and support the rights of its community by successfully listening to those who are laying complaints and resolving to assist them by bringing the perpetrators to the grinding wheels of justice. Encouraging and ensuring that police officials utilise the most modern means of investigation such as the DNA technology, provides the necessary balance to the exercise of professional discretion and heightened conviction rate by the police officials. The utilisation of biological evidence in investigation of rape cases is such a modern intervention – a way of providing insulation against internal and external interference with the proper and successful investigation of rape cases. / Forensic Investigation / M. Tech. (Forensic Investigation)

Democratic and accountable policing is one of the hallmarks of democracy. In a healthy democracy, a police service exists to protect and support the rights of its community by successfully listening to those who are laying complaints and resolving to assist them by bringing the perpetrators to the grinding wheels of justice. Encouraging and ensuring that police officials utilise the most modern means of investigation such as the DNA technology, provides the necessary balance to the exercise of professional discretion and heightened conviction rate by the police officials. The utilisation of biological evidence in investigation of rape cases is such a modern intervention – a way of providing insulation against internal and external interference with the proper and successful investigation of rape cases. / Forensic Investigation / M. Tech. (Forensic Investigation)

The open nature of Cell-Free Protein Synthesis (CFPS) systems has enabled flexible design, easy manipulation, and novel applications of protein engineering in therapeutic production, biocatalysis, and biosensors. This dissertation reports on three advances in the application of CFPS systems for 1) improving biocatalysis performance in industrial applications by site-specific covalent enzyme immobilization, 2) expressing and optimizing a difficult to express a mammalian protein in bacterial-based CFPS systems and its application for cost-effective, on-demand biosensors compatible with human body fluids, and 3) streamlining the procedure of an E. coli extract with built-in compatibility with human body fluid biosensors. Site-specific covalent immobilization stabilizes enzymes and facilitates recovery and reuse of enzymes which improves the net profit margin of industrial enzymes. Yet, the suitability of a given site on the enzyme for immobilization remains a trial-and-error procedure. This dissertation reports the reliability of several design heuristics and a coarse-grain molecular simulation in predicting the optimum sites for covalent immobilization of a target enzyme, TEM-1 ?-lactamase. This work demonstrates that the design heuristics can successfully identify a subset of favorable locations for experimental validation. This approach highlights the advantages of combining coarse-grain simulation and high-throughput experimentation using CFPS to efficiently identify optimal enzyme immobilization sites. Additionally, this dissertation reports high-yield soluble expression of a difficult-to-express protein (murine RNase Inhibitor or m-RI) in E. coli-lysate-based CFPS. Several factors including reaction temperature, reaction time, redox potential, and presence of folding chaperones in CFPS reactions were altered to find suitable conditions for m-RI expression. m-RI with the highest activity and stability was used to develop a lyophilized CFPS biosensor in human body fluids which reduced the cost of biosensor test by ~90%. Moreover, an E. coli extract with RNase inhibition activity was developed and tested which further streamlines the production of CFPS biosensors compatible with human body fluids.