One of the challenges in the 21st Century is to overcome the recalcitrance of lignocellulose for the production of liquid biofuels. Lignin is one of the key factors in this recalcitrance. Grasses such as Miscanthus and switchgrass could become major sources of lignocellulose. Barley has potential as a genetically-tractable research model for such novel bioenergy crops and also as a bioenergy crop itself. This thesis concerns the 4CL and the CCR enzymes on the lignin pathway which were chosen as the targets to manipulate lignin in barley. They were selected because there is evidence that suppression of each of them in dicot species can lead to increased saccharification. The 4CL and CCR genes constitute multigene families where members have different expression patterns. RNAi was used to down-regulate 4CL1 and CCR1 using a constitutive promoter via Agrobacterium-mediated transformation of barley. From an extensive screen of the primary transformants for changes in protein level and lignin content, six CCR and four 4CL lines were taken forward for detailed analysis. Antibodies were also raised against barley 4CL and CCR recombinant proteins and these showed substantial reductions in the respective target protein levels in the RNAi lines. Both 4CL and CCR transgenic lines had significant reductions in lignin content, and CCR lines had changes in lignin structure due to changes in the proportions of acid soluble and acid insoluble lignin. No substantial consistent adverse effects on key agronomic traits were apparent in the 4CL and CCR transgenics. Selected 4CL and CCR transgenics had improved saccharification yield after using three different pretreatment methods, which is a desirable feature for biofuel production.

Introdução: apesar do crescimento exponencial do interesse na pesquisa do zumbido, dos avanços científicos e do aprimoramento das técnicas terapêuticas observados nas duas últimas décadas, ainda é praticamente um consenso o fato da cura ser um objetivo inatingível. A ausência de estudos relativos à total remissão do zumbido e a escassez de discussões sobre o assunto revelam a descrença dos profissionais, o que funciona como reforço negativo para os pacientes que sofrem e almejam a cura. Objetivo: o presente estudo propõe a busca e a entrevista de indivíduos que já alcançaram a remissão total do zumbido para identificar possíveis características comuns do paciente, do zumbido ou do tipo de tratamento realizado. Método: Foram realizadas as seguintes estratégias de busca de participantes: 1) análise de banco de dados; 2) busca pelos parentes de pacientes que mencionam a cura; 3) divulgação da pesquisa em aulas ministradas a profissionais; 4) divulgação na internet. Foram incluídos os seguintes indivíduos: a) de qualquer sexo, idade, escolaridade ou nível socio-econômico; b) que tenham apresentado zumbido diariamente por pelo menos três meses em qualquer época da vida; c) que tenham apresentado remissão total do zumbido por pelo menos 6 meses; d) em casos de reaparecimento do zumbido, este deveria durar no máximo dois dias e ser desencadeado ou associado a algum evento claro. Foram excluídos os indivíduos que não responderam aos nossos contatos após três tentativas e aqueles que se encontravam em grau de habituação ou mascaramento do zumbido, mas que ainda o percebiam Resultados: a amostra final foi de 63 participantes. A idade variou de 14 a 89 anos (Média=52,69; Dp=16,33) e com predomínio do gênero feminino (68,25%). As características mais comuns do zumbido antes da remissão total foram: a) localização bilateral (52,31%); b) percepção diária de sons do tipo tom puro (57,14%); c) média de incômodo na escala visual analógica (0 a 10) de 7,57 e Dp 2,43, sendo 61,9% dos casos com incômodo alto (8 a 10) e apenas 22,22% sem limitação das atividades diárias; d) duração média de 49,07 meses (variância de 5.91), sendo 55,56% de casos crônicos (duração superior a 12 meses) e 44,44% de casos subagudos (duração de 4 a 12 meses). O número de tratamentos realizados até a remissão foi predominantemente de 0 a 2 (58,73%). O tipo de tratamento mais comum responsável pelo sucesso foi medicação (36,51%). Dentre os 57 dos 63 participantes que haviam realizado audiometria tonal, 47,62% tinham limiares normais. O tempo médio de remissão total do zumbido em meses foi de 92,03 (Dp 99,24). Conclusões: os pacientes que alcançaram remissão total do zumbido foram prioritariamente do gênero feminino, com percepção de sons do tipo tom puro, com nota de incômodo alta, audiometria normal bilateral e com poucas tentativas de tratamento. A localização (uni ou bilateral) e o tempo de duração do zumbido (subagudo ou crônico) não influenciaram a obtenção da remissão total. Apesar do incômodo alto e da repercussão na qualidade de vida da maioria dos pacientes, parece haver um subtipo de zumbido mais propenso à remissão total e duradoura com a instituição de um tratamento / Introduction: Despite the exponential growth of interest in tinnitus research, besides scientific advances and improvements in therapeutic techniques observed in last two decades, it is still a consensus that the total remission is an unattainable goal until now. The absence of studies on tinnitus remission and the scarcity of discussions on this subject reveal the practitioners\' disbelief, what represents a negative reinforcement for patients who suffer and seek the cure Objective: the present study proposes to search and interview individuals who have experienced the total remission of tinnitus in order to identify possible common features about patients, tinnitus or treatments. Method: The following strategies to search candidates were used: 1) database analysis; 2) search for the relatives of patients who mention the cure; 3) divulgation of the research in events for health professionals; 4) announcements on the web. The following individuals were included: a) of any gender, age, education or socioeconomic level; b) those who had tinnitus for at least three month period at any time of life; c) those who have presented tinnitus total remission for over 6 months; d) in cases of tinnitus recurrence, this should last no more than two days and had been triggered by some clear event. Subjects who did no respond to at least tree attempts of contact and those found in any degree of habituation or masking of tinnitus were excluded. The final sample was composed of 63 participants. The age varied from 14 to 89 years (mean = 52.69, SD = 16.33), female gender predominated (68.25%). The most common characteristics of tinnitus before remission were: a) bilateral location (52.31%); b) daily perception of a pure tone (57.14%); c) average score at visual analogue scale (0 to 10) of 7.57 and SD 2.49; 61.9% of cases with high discomfort (8 to 10) and only 22.22% without limitation of daily activities; d) average duration of 49.07 months (variance of 5.91), 55.56% of chronic cases (lasting more than 12 months) and 44.44% of subacute cases (duration of 4 to 12 months). The number of treatment attempts needed to reach total remission was 0 to 2 (58.73%). The most common type of treatment was medication (36.51%). Of the 57 participants who had tonal audiometry, 47.62% had normal thresholds. So far, the average duration of the total remission was 92,03 months (SD 99,24). Conclusions: patients who achieved total remission of tinnitus were primarily female, perceiving a tonal type of sound, with a high level of discomfort, with symmetrically normal audiometry and limited treatment attempts. The location (bilateral or unilateral) and duration of tinnitus (subacute or chronic) did not influence total remission. Despite of the severe repercussion on patient\'s quality of life, there seems to be a tinnitus subtype more prone to total and long-term remission after the institution of a certain treatment

O trato olivococlear medial realiza o controle eferente das células ciliadas externas, regulando as contrações lentas e atenuando as contrações rápidas. Com o surgimento das técnicas de pesquisa da amplitude das emissões otoacústicas sem e com estimulação acústica contra, ipsi ou bilateralmente, é possível estimar clinicamente as condições fisiológicas desse trato, uma vez que o efeito resultante de redução/supressão das emissões otoacústicas indica seu funcionamento efetivo. O envelhecimento implica em diminuição da atividade do sistema auditivo central, em função da degeneração das estruturas envolvidas nas habilidades auditivas. A perda de fibras axonais e a diminuição da bainha de mielina nas estruturas do trato representam perda da eficácia de sua função. O objetivo deste estudo foi investigar o efeito da idade na atividade do trato sobre as propriedades mecânicas da cóclea, por meio da análise da amplitude das emissões otoacústicas com estimulação acústica contralateral. A casuística foi composta por 75 indivíduos ouvintes, de ambos os sexos, agrupados conforme a idade. A metodologia para captação das emissões otoacústicas foi o modo convencional, com clique linear na intensidade de 60 dB NPS e o ruído branco, como estímulo contralateral, em 60 dB NPS. A análise dos resultados considerou o valor do response das orelhas e a comparação entre os grupos etários. Os resultados revelam diferenças estatisticamente significantes entre os valores do response das EO sem e com estimulação acústica contralateral, nos indivíduos de 20 a 39 anos. O efeito redução/supressão das EOE-T diminui substancialmente com a idade, por volta da quarta década. Conclui-se que o envelhecimento prejudica a efetividade da atividade do trato sobre a cóclea. / Medial olivocochlear tract makes the efferent control of the outer hair cells, controlling the slow contractions and mitigating rapid contractions. Upon the appearance of techniques for the investigation of the amplitude of otoacoustic emissions with and without acoustic stimulation against, ipsi and bilaterally, it is now possible to stimulate clinically the physiologic conditions of this tract, once the resulting effect of reduction/suppression of otoacoustic emissions indicates its effective action. Aging brings a decrease of the activity of central auditory system, as a function of the degeneration of structures involved in auditory abilities. The loss of axon fibers and the decrease of myelin sheath in tract structures causes the loss of efficacy in its function. It was the objective of this study to assess the effect of aging in tract activity upon mechanical properties of cochlea, through the analysis of the amplitude of acoustic emissions with a contralateral acoustic stimulation. Casuistic comprised 75 both sex hearing individuals, grouped according to age. Methodology to capture otoacoustic emissions employed conventional means, with a linear click at a 60 dB NPS intensity and white sound, as a contralateral stimulus, at 60 dB NPS. The analysis of results considered the value of the response of the ears and the comparison of between age groups. Results show statistically significant differences between values for response of EO with and without a contralateral acoustic stimulation, in individuals between 20 to 39 years of age. The reduction/suppression effect of EOE-T decreases greatly with aging, around the fourth decade. It is concluded that aging impairs the efficacy of the activity of tract upon the cochlea.

The objective of this study was to identify genes that are differentially regulated during diapause when compared with nondiapausing pupae in Sarcophaga crassipalpis. The results of a Suppression Subtractive Hybridization procedure was used to indentify genes that are differentially regulated in both diapause and nondiapausing states while suppressing genes that are common to both states. Randomly picked colonies from both subtractive libraries were isolated and the inserts sequenced. The sequences were analyzed using the bioinformatics tools NCBI, BlastX, Clustal W, etc. Out of 384 clones, 59 genes were found to be upregulated during diapause and 37 genes were found to be upregulated during a nondiapause pupal stage, no genes were found to be expressed commonly in both the diapause and nondiapause constructed libraries.

No description available.

No description available.

Auditory recognition memory in non-human primates is not well understood. Monkeys have difficulty acquiring auditory memory tasks, and limited capability maintaining auditory information over memory delays, relative to studies of visual memory. Neural substrates of auditory discrimination and recognition memory depend on superior temporal gyrus (STG), instead of rhinal cortex necessary for visual memory (Fritz et al., 2005). The current project assessed behavioral and neural correlates of auditory processing and memory function in monkeys, particularly focusing on the dorsal temporal pole (dTP), the rostral portion of STG. Chapter 2 examined recognition memory of monkeys under influences of various sound types. In a delayed matching-to-sample (DMTS) task, rhesus monkeys were trained to determine if two sounds, separated by a 5-second delay, were same (match trials) or different (nonmatch trials). Results demonstrated monkey vocalizations served as better cues than other sound types for auditory memory performance. Memory improvements may be due to familiarity and biological significance of con-specific sounds, analogous to using facial stimuli during visual tasks. Chapter 3 examined neuronal activity of dTP, when two monkeys performed an auditory DTMS task and listened to sound stimuli. Population encoding of sample stimuli in dTP was closely associated with memory accuracy. Moreover, a suppression effect on identical sounds was present, similar to processing in the ventral visual processing stream, inferior temporal cortex (ITC) and ventral temporal pole (vTP). Delay-related activity of dTP was weak, limited and short-lived, in contrast to visual studies reporting sustained activity over memory delays in ITC, vTP and prefrontal cortex. The findings provide preliminary evidence on why monkeys show limited memory capability, compared to visual memory, for auditory information. Neurons of dTP were sound-selective, and mainly evoked by one to four discrete stimuli only. Sound types and simple acoustic properties of sound stimuli cannot completely account for response profiles of dTP neurons. The findings suggest dTP is a higher order auditory area, and receives information from various auditory areas along STG. Dorsal temporal pole fits into proposals of neural networks for auditory processing, in which a hierarchical organization of information flow exists within the primate auditory nervous system.

Although tic disorders are diagnosed as neurological disorders, neurobehavioral models suggest that tics are controlled by premonitory urges that may be conditioned to become aversive through childhood, and that tics are exhibited to alleviate such phenomena. However, only indirect measures have been used to assess the presence of the premonitory urge. This study utilized self-report and GSR measurements to examine whether a punishing contingency conditioned stimuli to be aversive during conditions of tic suppression and whether punishing contingencies exacerbate aversive private phenomena in two adults. Results indicated that conditions of response cost (RC) and differential reinforcement (DRO) were effective at reducing the number of tics compared to baseline. Moreover, GSR was unrelated to urge and suppression conditions despite higher self-reported urge ratings in DRO and RC conditions. Implications of findings are discussed.

The Westboro Baptist Church and Phelps family are notorious for their signs proclaiming anti-homosexuality epithets such as “God Hates Fag,” and references to Biblical verses to revile homosexuality (e.g., Romans 1:24-27). Although the homonegativity bias that Westboro patrons and many traditional Christians express is often understood as being rooted in religion, I proposed the possibility that religion can be a justification rather than source of homonegativity. That is, although religion typically is seen as the source of prejudice towards LGBT+ people, I argued that this relationship might work in reverse. I examined under which conditions this “reverse” phenomena might occur in both the Pilot Study and follow-up study. My Pilot Study results suggested that there are differences in participants’ level of Religiosity after falsely being told they held LGBT+ prejudices. These differences were moderated by political alignment. Relatively liberal participants were more likely to have higher levels of Religiosity after receiving Biased Feedback compared to those who received Neutral Feedback. In terms of those who were more politically conservative, their level of Religiosity tended to be about the same regardless of which feedback they received. When it came to Biblical Literalism, however, relatively conservative participants had higher levels of literalism when given the Neutral Feedback compared to relatively liberal participants when given the same Feedback. These findings were mostly replicated in the follow-up Main Study. The results from both studies suggest that, under some circumstances, people might use religion to justify their prejudice towards LGBT+, and the strategy they use is affected by their political alignment (liberal or conservative). There could be differences in motivations between people who are more politically conservative and those who are more politically liberal. These potential differences are addressed in the discussion.

Breast cancer is the most common non-skin cancer in women and the second most common cause of cancer-related death in U.S. women. Despite numerous advances in treatment strategies against breast cancer, the presence of undetected distant metastasis of the primary tumor remains the main cause of mortality. Current screening and detection methods such as mammograms are simply not sensitive enough to detect formation of metastasis. Further, currently available therapies against metastatic breast cancer do not provide a complete cure for the disease. Thus, understanding the biology and molecular factors involved in cancer metastasis will help aid in preventing the onset of metastasis and discovering an effective treatment for this deadly disease. My research focused on understanding the mechanism of action of one such factor, breast cancer metastasis suppressor 1 (BRMS1), a suppressor gene found deleted in late stage breast cancers. The goal of my dissertation was to investigate the role of membrane signaling lipids phosphoinositides, specifically phosphatidylinositol(4,5)bisphosphate (PI(4,5)P2) in BRMS1-mediated metastasis suppression in MDA-MB-435 and MDA-MB-231 human breast carcinoma cells. My studies revealed BRMS1 selectively reduced receptor tyrosine kinases (RTK) and Gprotein coupled receptors (GPCR) expression and downstream signaling in human breast carcinoma cells. My observations are critical as many of these receptors are upregulated in metastatic breast cancer and PI(4,5)P2 is a critical constituent for mediating their downstream signaling events. Further, using immunoblotting studies, I uncovered a possible compensatory mechanism in tumor cells to overcome downregulation of PI(4,5)P2 by BRMS1 and maintain its downstream signaling. When studied for BRMS1 regulation of enzymes involved in PI(4,5)P2 synthesis, I showed BRMS1 completely inhibits phosphatidylinositol 4-phosphate 5-kinase β (PIP5Kβ) expression. Using overexpression studies, I showed PIP5Kβ to be the major contributor to the cellular PI(4,5)P2 pool required for agonist-induced intracellular calcium rise. Taken together, my dissertation research has identified some critical breast cancer markers and revealed signaling pathways altered by BRMS1 in human breast carcinoma cells that can be studied as potential therapeutic targets against breast cancer metastasis.