<p>Forebrain cholinergic projection systems innervate the entire cortex and hippocampus. These cholinergic systems are involved in a wide range of cognitive and behavioral functions, including learning and memory, attention, and sleep-waking modulation. However, the <italic>in vivo</italic> physiological mechanisms of cholinergic functions, particularly their fast dynamics and the consequent modulation on the hippocampus and cortex, are not well understood. In this dissertation, I investigated these issues using a number of convergent approaches.</p><p> First, to study fast acetylcholine (ACh) dynamics and its interaction with field potential theta oscillations, I developed a novel technique to acquire second-by-second electrophysiological and neurochemical information simultaneously with amperometry. Using this technique on anesthetized rats, I discovered for the first time the tight <italic>in vivo</italic> coupling between phasic ACh release and theta oscillations on fine spatiotemporal scales. In addition, with electrophysiological recording, putative cholinergic neurons in medial setpal area (MS) were found with firing rate dynamics matching the phasic ACh release. </p><p> Second, to further elucidate the dynamic activities and physiological functions of cholinergic neurons, putative cholinergic MS neurons were identified in behaving rats. These neurons had much higher firing rates during rapid-eye-movement (REM) sleep, and brief responses to auditory stimuli. Interestingly, their firing promoted theta/gamma oscillations, or small-amplitude irregular activities (SIA) in a state-dependent manner. These results suggest that putative MS cholinergic neurons may be a generalized hippocampal activation/arousal network. </p><p> Third, I investigated the hypothesis that ACh enhances cortical and hippocampal immediate-early gene (IEG) expression induced by novel sensory experience. Cholinergic transmission was manipulated with pharmacology or lesion. The resultant cholinergic impairment suppressed the induction of <italic>arc</italic>, a representative IEG, suggesting that ACh promotes IEG induction. </p><p> In conclusion, my results have revealed that the firing of putative cholinergic neurons promotes hippocampal activation, and the consequent phasic ACh release is tightly coupled to theta oscillations. These fast cholinergic activities may provide exceptional opportunities to dynamically modulate neural activity and plasticity on much finer temporal scales than traditionally assumed. By the subsequent promotion of IEG induction, ACh may further substantiate its function in neural plasticity and memory consolidation.</p> / Dissertation

Middle-line managers (MLMs) are becoming increasingly important for an organization. Coordination forms (immediate coordination and in-advance coordination) are used in the organization. In this thesis, four variables: organizational type, the degree of decentralization of organization, qualification and uncertainty are analyzed for investigating what are main factors influencing the use of coordination forms in Chinese organizations from MLM’s perspective.The empirical data for the study was gathered by a survey questionnaire, which involved a quota sampling of 158 MLMs in Chinese organizations. The interesting results justify the tendency to use in-advance coordination is greater for organizations that are characterized by a high level of uncertainty, especially learning and goal-setting; and the tendency to use in-advance coordination is greater for organizations that are characterized by a high level of decentralization.

Choosing with whom to mate is one of the most important decisions a female makes in her lifetime and inter-individual variation of these preferences can have important evolutionary consequences. In order to get a complete understanding of why and how females choose a mate, we must identify factors that can contribute to variation of female mate choice. Many decades of research sought to understand ultimate mechanisms of female mate choice with proximate mechanisms receiving a lot more attention in recent years. For my thesis, I identify intrinsic and extrinsic factors that correlate with individual variation of female Xiphophorus nigrensis mate preference. I provide evidence that a female’s size (e.g. age and sexual experience) as well as male behavioral displays can predict female mate preference. Using genes associated with female mate preference (neuroserpin, neurologin-3), I identify four brain regions (Dl, Dm, HV, POA) that show significant differences in gene expression between females exhibiting high preference for males relative to females displaying little mate preference. Neuroserpin and neuroligin-3 gene expression within these brain regions are also positively correlated with female mate preference behavior. Two of these brain regions (Dm and Dl) integrate multisensory information and are found in the putative teleost mesolimbic reward circuitry; the other two regions (HV and POA) are involved in sexual behaviors. With the implication of the reward circuitry, I assess whether there are changes in dopamine synthesis (via tyrosine hydroxylase, TH) in dopaminergic brain regions associated with the degree of mate preference. I do not find evidence of rapid changes (within 30 minutes) of TH expression (i.e. dopamine synthesis) in dopaminergic brain regions related to variation in female mate preference. Collectively my results suggest that mate preference behavior in the brain may be coordinated not just through regions associated with sexual response but also through forebrain areas that may integrate primary sensory information, with no associated changes of a proxy for dopamine synthesis in dopaminergic brain regions. / text

No description available.

Histone deacetylases (HDAC) 1 and 2 play crucial role in chromatin remodeling and gene expression regimes, as part of multiprotein corepressor complexes. Protein kinase CK2-driven phosphorylation of HDAC1 and 2 regulates their catalytic activities and is required to form the corepressor complexes. Phosphorylation-mediated differential distributions of HDAC1 and 2 complexes in regulatory and coding regions of transcribed genes catalyze the dynamic protein acetylation of histones and other proteins, thereby influence gene expression. During mitosis, highly phosphorylated HDAC1 and 2 heterodimers dissociate and displace from mitotic chromosomes. Our goal was to identify the kinase involved in mitotic phosphorylation of HDAC1 and 2. We postulated that CK2-mediated increased phosphorylation of HDAC1 and 2 leads to dissociation of the heterodimers, and, the mitotic chromosomal exclusions of HDAC1 and 2 are largely due to the displacement of HDAC-associated proteins and transcription factors, which recruit HDACs, from chromosomes during mitosis. We further explored the role of un- or monomodified HDAC1 and 2 complexes in immediate-early genes (IEGs), FOSL1 (FOS-like antigen-1) and MCL1 (Myeloid cell leukemia-1), regulation. Dynamic histone acetylation is an important regulator of these genes that are overexpressed in a number of diseases and cancers. We hypothesized that transcription dependent recruitment of HDAC1 and 2 complexes over the gene body regions plays a regulatory role in transcription and splicing regulation of these genes. We present evidence that CK2-catalyzed increased phosphorylation of HDAC1 and 2 regulates the formation of distinct corepressor complexes containing either HDAC1 or HDAC2 homodimers during mitosis, which might target cellular factors. Furthermore, the exclusion of HDAC-recruiting proteins is the major factor for their displacement from mitotic chromosomes. We further demonstrated that un- or monophosphorylated HDAC1 and 2 are associated with gene body of FOSL1 in a transcription dependent manner. However, HDAC inhibitors prevented FOSL1 activation independently of the nucleosome response pathway, which is required for IEG induction. Interestingly, our mass spectrometry results revealed that HDAC1 and 2 interact with a number of splicing proteins, in particular, with serine/arginine-rich splicing factor 1 (SRSF1). HDAC1 and 2 are co-occupied with SRSF1 over gene body regions of FOSL1 and MCL1, regardless of underlying splicing mechanisms. Using siRNA-mediated knockdown approaches and HDAC inhibitors, we demonstrated that alternative splicing of MCL1 is regulated by RNA-directed localized changes in the histone acetylation levels at the alternative exon. The change in histone acetylation levels correlates with the increased transcription elongation and results in change in MCL1 splicing by exon skipping mechanism. Taken together, our results contribute to further understanding of how the multi-faceted HDAC1 and 2 complexes can be regulated and function in various processes, including, but not limited to, transcription regulation and alternative splicing. This can be an exciting area of future research for therapeutic interventions.

No description available.

Histone deacetylases (HDAC) 1 and 2 play crucial role in chromatin remodeling and gene expression regimes, as part of multiprotein corepressor complexes. Protein kinase CK2-driven phosphorylation of HDAC1 and 2 regulates their catalytic activities and is required to form the corepressor complexes. Phosphorylation-mediated differential distributions of HDAC1 and 2 complexes in regulatory and coding regions of transcribed genes catalyze the dynamic protein acetylation of histones and other proteins, thereby influence gene expression. During mitosis, highly phosphorylated HDAC1 and 2 heterodimers dissociate and displace from mitotic chromosomes. Our goal was to identify the kinase involved in mitotic phosphorylation of HDAC1 and 2. We postulated that CK2-mediated increased phosphorylation of HDAC1 and 2 leads to dissociation of the heterodimers, and, the mitotic chromosomal exclusions of HDAC1 and 2 are largely due to the displacement of HDAC-associated proteins and transcription factors, which recruit HDACs, from chromosomes during mitosis. We further explored the role of un- or monomodified HDAC1 and 2 complexes in immediate-early genes (IEGs), FOSL1 (FOS-like antigen-1) and MCL1 (Myeloid cell leukemia-1), regulation. Dynamic histone acetylation is an important regulator of these genes that are overexpressed in a number of diseases and cancers. We hypothesized that transcription dependent recruitment of HDAC1 and 2 complexes over the gene body regions plays a regulatory role in transcription and splicing regulation of these genes. We present evidence that CK2-catalyzed increased phosphorylation of HDAC1 and 2 regulates the formation of distinct corepressor complexes containing either HDAC1 or HDAC2 homodimers during mitosis, which might target cellular factors. Furthermore, the exclusion of HDAC-recruiting proteins is the major factor for their displacement from mitotic chromosomes. We further demonstrated that un- or monophosphorylated HDAC1 and 2 are associated with gene body of FOSL1 in a transcription dependent manner. However, HDAC inhibitors prevented FOSL1 activation independently of the nucleosome response pathway, which is required for IEG induction. Interestingly, our mass spectrometry results revealed that HDAC1 and 2 interact with a number of splicing proteins, in particular, with serine/arginine-rich splicing factor 1 (SRSF1). HDAC1 and 2 are co-occupied with SRSF1 over gene body regions of FOSL1 and MCL1, regardless of underlying splicing mechanisms. Using siRNA-mediated knockdown approaches and HDAC inhibitors, we demonstrated that alternative splicing of MCL1 is regulated by RNA-directed localized changes in the histone acetylation levels at the alternative exon. The change in histone acetylation levels correlates with the increased transcription elongation and results in change in MCL1 splicing by exon skipping mechanism. Taken together, our results contribute to further understanding of how the multi-faceted HDAC1 and 2 complexes can be regulated and function in various processes, including, but not limited to, transcription regulation and alternative splicing. This can be an exciting area of future research for therapeutic interventions.

Baigiamojo darbo tikslas – nustačius tiesioginių emocijų daromą poveikį televizinės reklamos veiksmingumui, pasiūlyti, kaip jas būtų galima panaudoti didinant televizinės reklamos veiksmingumą. Teorinėje darbo dalyje analizuojami moksliniai šaltiniai ir empiriniai tyrimai, susiję su emocijų poveikiu reklamos suvokimui, veiksmingumui ir efektyvumui, taip pat tiesioginių emocijų, tai yra su pačia reklama nesusijusių, poveikis pastariesiems veiksniams. Analitinėje baigiamojo darbo dalyje pateikiami ir analizuojami eksperimentinio tyrimo rezultatai, atlikto dirbtinai iššaukiant eksperimentines tiesiogines atsitiktines ir integruotąsias emocijas prieš žiūrint televizijos reklaminį intarpą ir tyrimo rezultatus lyginant su kontrolinės grupės rezultatais. Projektinėje darbo dalyje suformuluoti sprendimai, pritaikomi reklamos užsakovams ir transliuotojams. Pasiūlymai apima: • Skirtingo tiesioginių emocijų poveikio panaudojimą skirtingiems reklamos tikslams pasiekti, • Reklamos klipo demonstravimo laiko pasirinkimo planavimą, • Tiesioginių emocijų tyrimo sukūrimą ir kainą Lietuvoje. / The objective of diploma paper – to find the effect of immediate emotions on the efficiency of television advertisement; and to suggest how they could be employed to increase the efficiency. In the theory part of the diploma paper – the analysis of the scholar papers and academic researches is conducted, regarding the impact of emotions on advertisement understanding, efficiency and effectiveness and also the immediate emotions, which are unconnected to advertising, but still effecting its understanding. In the analytic part of the diploma paper the data of experimental research results is presented and analyzed. The research was conducted by artificially summoning the immediate accidental and integrated emotions before watching the television advertising and comparing the results with control group results. In the project part the solutions for the advertising customers and broadcasters are made. The solutions cover: • The different effects of the immediate emotions using for different goals of advertisement, • The planing of the time of advertisement placement in television program, • The creation of immediate emotions research and cost in Lithuania.

<p>Estradiol modulates the use of spatial navigation strategies in female rats. The presence of circulating estradiol enhances learning on tasks that require the use of a hippocampus-dependent place strategy and impairs learning on tasks that require the use of a dorsal striatum-dependent response strategy. When either strategy may be used successfully, estradiol biases females to use a place strategy. While this behavioral effect has been well-described in the young adult female rat, little is known about the mechanisms in the brain that underlie it or how it changes across age. The experiments in this dissertation examined how age, previous experience, and hormonal condition affect the ability of estradiol to modulate learning during explicit training of place and response tasks, as well as navigation strategy use during ambiguous navigation tasks. Age highly influenced the ability of estradiol to influence strategy use. While female rats could use place and response strategies to navigate by postnatal day (PD) 21, estradiol did not bias them to use a response strategy until PD26, just before puberty. In adulthood, previous navigation experience and estradiol interacted to influence navigation strategy use on a series of experiences to an ambiguous navigation task. And, estradiol impaired learning during explicit response training but did not affect place learning. In middle age, estradiol further impaired response learning but still did not affect place learning. Long-term hormone deprivation, however, was detrimental to acquisition of a place task but did not affect response learning. These experiments also examined the effects of estradiol on activity, plasticity, and reliability of neuronal ensembles in several subregions of the hippocampus and striatum during spatial navigation using cellular and molecular techniques that take advantage of the kinetics of the immediate-early genes c-fos and Arc. Increased activation and plasticity during active exploration across several subregions of the hippocampus and striatum reflected similar inputs to these neural systems and similar effects of exploration. However, estradiol modulated the plasticity and reliability of neuronal ensembles in the hippocampus and striatum specifically during goal-directed spatial navigation. Estradiol increased plasticity in CA1 of all behaviorally-trained rats, but only place strategy users displayed high reliability in this plasticity across training and probe trials on a navigation task. Estradiol prevented increase in plasticity and reliability in the dorsolateral striatum displayed by low estradiol response strategy users. These experiments reveal how several factors, including age, influence estradiol's modulation of spatial navigation strategy use and suggest functional mechanisms by which this modulation occurs.</p> / Dissertation

Herewith are presented the results of an investigation the statistical power of USP compendial release tests and recommended alternatives. &lt;br&gt;The U.S. drug supply chain, formerly protected by a closed distribution network, is now threatened by the legal and illegal importation of drug products. Whereas quality can never be inspected into final products, compendial release standards may represent the only valid assessment that products of dubious origin would receive. Reliable tests for content uniformity and dissolution are required to protect the safety of the supply chain. A study was designed to test the hypothesis that existing compendial tests for content uniformity and dissolution would protect the supply chain against substandard and counterfeit drugs if basic field tests failed. &lt;br&gt;Compendial tests for content uniformity and dissolution were evaluated for statistical power using simulation studies. The results revealed that the revised content uniformity test, based on tolerance analysis, was subject to an unacceptable level of consumers' risk. The Bergum method proved to be an excellent secondary standard for product assessment and is recommended as an alternative to the USP method. Simulations with the USP dissolution test revealed significant weaknesses and inconsistencies in the test structure. Theoretical models and power assessments confirmed that the coverage specification of the dissolution test was an unacceptably high 50% coverage with 50% confidence. &lt;br&gt;A Bayesian D-optimal design program was used to investigate alternative methods to improve the coverage capability of the USP dissolution test. The result of this program was the identification of two alternatives to the existing USP procedure. The first alternative is based on the addition of attribute coverage tests to stages 2 and 3 of the USP test, whereas the second alternative is based on the concept of tolerance analysis. &lt;br&gt;Validation studies confirmed that both alternatives significantly improved the statistical power of the USP dissolution test without increasing the sample size or modifying the current three-stage procedure. The attribute test is non-parametric and behaves similarly to the existing USP with improved coverage, whereas the continuous alternative is more sensitive and is consistent with the recent revisions to the content uniformity test. / Mylan School of Pharmacy and the Graduate School of Pharmaceutical Sciences / Pharmaceutics / PhD / Dissertation