Thesis (Ph. D.)--University of Washington, 1997. / Vita. Includes bibliographical references (leaves [114]-130).

Thesis (Ph. D.)--University of Washington, 1997. / Vita. Includes bibliographical references (leaves [64]-71).

Thesis (Ph.D.)--University of Florida, 2005. / Typescript. Title from title page of source document. Document formatted into pages; contains 165 pages. Includes Vita. Includes bibliographical references.

Made available in DSpace on 2015-07-22T20:49:56Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-11-26. Added 1 bitstream(s) on 2015-08-11T03:25:31Z : No. of bitstreams: 1 Publico-00387.pdf: 2025016 bytes, checksum: 1d807dcd8112ef82a43a87d394716763 (MD5) / Objetivos: 1- Investigar o efeito imediato (01 sessão) e mediato (10 sessões) da acupuntura no tratamento da síndrome de apnéia-hipopnéia obstrutiva do sono (SAHOS) de grau moderado. 2- Avaliar a especificidade da técnica, comparando acupuntura manual, eletroacupuntura 2 Hz e eletroacupuntura 10 Hz. 3- Avaliar o envolvimento da serotonina neste mecanismo de ação da acupuntura. Método: Um ensaio clínico, randomizado, controlado, simples cego, foi conduzido no Departamento de Fisiologia, no Setor de Distúrbios do Sono, no Setor de Medicina Chinesa-Acupuntura e no Laboratório de Rim e Hormônios da Universidade Federal de São Paulo - Escola Paulista de Medicina, no período de Janeiro de 2004 a outubro de 2006. Foram recrutados 43 pacientes previamente diagnosticados clínica e polissonograficamente como portadores de SAHOS moderada {índice de apnéia-hipopnéia (IAH) > 15/hora < 30/hora}. Os pacientes foram randomizados em quatro grupos: grupo controle (CTRL), grupo acupuntura manual, grupo eletroacupuntura 2 Hz e grupo eletroacupuntura 10 Hz. O grupo controle não recebeu qualquer tipo de intervenção. Os grupos acupuntura manual e eletroacupuntura receberam tratamento por 05 semanas (2 aplicações por semana). Todos os pacientes foram avaliados pré e pós-tratamento através de exame polissonográfico e da dosagem de serotonina. A dosagem de serotonina foi feita por HPLC-ED (Cromatografia Líquida acoplada a Detecção Eletroquímica). Resultados: Em relação ao efeito imediato (01 sessão) da acupuntura, houve diminuição significativa do IAH (P=0,006; P=0,009), dos eventos respiratórios (P=0,012; P=0,005), dos microdespertares (P=0,046; P=0,05) e da saturação de O2 (P=0,006; P=0,035) nos grupos acupuntura manual e eletroacupuntura 10 Hz, antes e depois de uma aplicação de acupuntura. O índice de apnéia (P=0,028) e o índice de hipopnéia (P=0,046) tiveram diminuição significativa apenas no grupo eletroacupuntura 10 Hz. Não houve alterações significativas nos grupos eletroacupuntura 2 Hz e grupo controle quando foram comparados polissonograficamente antes e após a sessão de acupuntura. Quando todos os grupos foram comparados ao grupo controle, apenas o grupo eletroacupuntura 10 Hz apresentou diminuição significativa do IAH. Em relação ao efeito mediato (10 sessões) da acupuntura houve diminuição significativa do tempo de início do sono (P=0,011; P=0,005), do IAH (P=0,011; P=0,006), do índice de apnéia (P=0,042; P=0,005) dos eventos respiratórios (P=0,011; P=0,009) e dos microdespertares (P=0,011; P=0,021) nos grupos acupuntura manual e eletroacupuntura 10 Hz, antes e depois do tratamento. O índice de hipopneia (P=0,021) teve diminuição significativa apenas no grupo eletroacupuntura 10 Hz. O tempo de inicio do sono (P=0,005; P=0,005) dimimuiu significativamente tanto no grupo controle quanto no grupo eletroacupuntura 2 Hz. A exceção deste dado, nenhum outro evento se modificou significativamente nestes grupos. Quando todos os grupos foram comparados ao grupo controle, apenas o grupo eletroacupuntura 10 Hz apresentou diminuição significativa do IAH e dos microdespertares (P=0,004; P=0,008). Em relação ao envolvimento da serotonina no mecanismo de ação imediato da acupuntura na SAHOS Moderada, não observamos significância estatística entre grupos quando comparados ao Controle: Acupuntura Manual (P=0,7674), Eletroacupuntura 2 Hz (P=0,9849) e Eletroacupuntura 10 Hz (P=0,1042). Em relação ao envolvimento da serotonina no mecanismo de ação mediato da acupuntura na SAHOS Moderada o grupo Acupuntura Manual apresentou significância estatística quando comparado ao grupo Controle (P=0,0067) e o grupo Eletroacupuntura 10 Hz, apresentou tendência à significância estatística quando comparado ao grupo Controle (P=0,0984). A não significância estatística destes resultados parece estar relacionada à variabilidade inicial da 5HT e ao pequeno tamanho da amostra. Conclusões: A acupuntura manual e a eletroacupuntura 10 Hz são mais eficazes que a eletroacupuntura 2 Hz em melhorar o IAH e outros eventos respiratórios do sono de pacientes portadores de SAHOS Moderada. Esta eficácia foi observada tanto no efeito imediato (01 sessão de acupuntura), como no efeito mediato (10 sessões de acupuntura). Os resultados preliminares relacionados à serotonina, já apontam para uma direção neurofisiológica que possa justificar a eficácia da acupuntura no tratamento de pacientes portadores da SAHOS. / STUDY OBJECTIVES: 1 - To investigate the immediate (single session) and mediate (ten sessions) effect of acupuncture over the sleep pattern of patients with moderate obstructive sleep hypopnea apnea (OSHA). 2 - To compare manual acupuncture (MA) and electroacupuncture (EA) with different stimulation frequencies (2 Hz and 10 Hz). 3 - To determine the role of endogenous serotonin in this mechanism. SETTING: Department of Physiology, Sleep Division of Department of Psychobiology, Chinese Medicine Division and Department of Nephrology, Federal University of São Paulo, Brazil. INTERVENTIONS: Acupuncture applied in true acupoints accompanied by needle manipulations or electroacupunture 2 hz and 10 Hz for 30 minutes in all patients, except control group pacients. DESIGN: In a randomized, placebo-controlled and single blind study, fourty-three patients with an apnea-hypopnea index (AHI) of 15 to 30 per hour were randomly assigned to 4 groups: control group, MA group, EA 2Hz group and EA 10Hz group. The control group received no intervention. The MA group, EA 2Hz group and EA 10Hz group received treatment for five weeks (02 acupuncture aplications per week). All the patients were evaluated pre-treatment and pos-treatment by polissomnography recordings and assessment of serotonin levels. The 5HT measurement was made with a HPLC-ED (Liquid Chromatografy coupled with Electrochemical Detection). MEASUREMENTS AND RESULTS: 1 – Immediate effect: when compared before and after treatment, the AHI (P=0,006; P=0,009), respiratory events (P=0,012; P=0,005), microarousals (P=0,046; P=0,05) and mean O2 saturation (P=0,006; P=0,035) showed significant improvement in both the manual and electroacupuncture 10 Hz respectively. The apnea index (P=0,028) and the hipopneia index (P=0,046) showed significant improvement only in EA 10 Hz group. There were no significant changes in the electroacupuncture 2 Hz as well as in the control group when compared before and after (first and second for controls) PSG recordings. When compared each group and control group, AHI significantly decreased only in the EA 10 Hz. 2 – Mediate effect: When compared before and after treatment, sleep onset (P=0,011;P=0,005), AHI (P=0,011;P=0,006), apnea index (P=0,042;P=0,005), respiratory events (P=0,011;P=0,009) and microarousals (P=0,011;P=0,021 ) significantly improved in MA and EA 10 Hz respectively. Hipopnea-index (P=0,021) decreased only in the EA 10Hz. Latency for sleep onset (P=0,005;P=0,005) decreased in both EA 2Hz and control group. Except for this change, there were no other significant changes in EA 2 Hz, or in control group after treatment. When compared to the control group the only significant differences were AHI and microarousals that were decreased in EA 10 Hz group (P=0,004;P=0,008). As concerned to the levels of seric serotonin we could not evidence an involvement of this neurotransmitter (as measured in blood) with the immediate effects of acupuncture for OSHA (MA vs. controls [P=0,7674], EA 2 Hz vs controls [P=0,9849]) despite a tendency for a difference for EA 10 Hz as compared to controls (P=0,1042). With regard to an effect of mediate acupuncture in moderate OSHA we found that seric serotonin levels were signficantly different form controls for MA (P=0,0067) with a tendency for group EA 10 Hz (P=0,0984). The lack of more robust differences for seric serotonin levels might have been influcenced by a smaller numeber of subjects as that required for this sort of evaluations. Conclusions: MA and EA 10 Hz have greater efficacy as compared to EA 2 Hz in improving AHI and other respiratory events in patients with moderate OHSA. This effect was shown to hold for both in the immediate (1 acupuncture session) as in the mediate (10 sessions) acupuncture application. The preliminary serotonin results indicate a possible mechanistic link between acupuncture treatment and its beneficial effects for the treatment of patients with moderate OHSA. / TEDE / BV UNIFESP: Teses e dissertações

The concerns of the present study were to determine 1) the conditions necessary to measure displaceable [³H] L-aspartate binding to membrane fractions of the rat brain, 2) whether the binding demonstrated the charcteristics of the site which is active in vivo, and 3) whether the acidic amino acid neurotransmitters aspartate and glutamate bind to identical or different sites by comparing the pharmacological specificities of the [³H] L-aspartate binding with that of [³H] L-glutamate. The conditions of the [³H] L-aspartate binding assay were determined in synaptosomal and total particulate fractions of whole rat brain. The reaction mixture which included the membrane fraction suspended in Tris-HCl buffer (pH 7.4) in the presence or absence of the compound under test, was incubated at 37°C for 30 minutes. The reaction was stopped by centrifugation and the radioactivity in the pellet counted by liquid scintillation spectrometry. The [³H] L-aspartate binding was characterized in total particulate fractions of rat cerebellum. The apparent dissociation constant (K[sub=D]) and maximum binding (Bmax), as determined by Scatchard analysis, are 1.64 ± 0.34 μM and 7711 ± fmol/mg protein respectively. The displaceable binding is reversible, saturable, independent of the presence of NA⁺, has an affinity in the range where the neurotransmitter is active in vivo, and demonstrates a pharmacological specificity which includes stereospecificity. The compounds tested to demonstrate the pharmacological specificity were L-aspartate (IC[sub=50] = 1.81 μM), D-aspartate (IC[sub=50] = 46.6 μM), L-glutamate (IC[sub=50] = 1.24 μM), N-methyl-DL-aspartate (inactive), kainate (inactive), D-alpha-aminoadipate (inactive), and L-alpha-aminoadipate (IC[sub=50] =7.12 μM). The pharmacological specificity of [³H] L-aspartate binding was different from that of [³H] L-glutamate. When the binding data only are considered, therefore, separate receptors for aspartate and glutamate are indicated. The pharmacological specificity of the [³H] L-aspartate binding, that is the affinity of the binding site for N-methyl-DL-aspartate, D- and L-alpha-aminoadipate, however, does not correlate with the potency of these compounds derived from iontophoretic studies. L-alpha-aminoadipate is very effective while N-methyl-DL-aspartate and D-alpha-aminoadipate do not displace the [³H] L-aspartate binding. In iontophoretic studies, N-methyl-D-aspartate and D-alpha-aminoadipate are very potent as compared to aspartate while L-alpha-aminoadipate Is inactive. The [³H] L-aspartate binding then may not represent the site which is active in vivo. The characteristics of the aspartate site in vivo, however, may not be truely represented in iontophoretic studies because of, for example, uptake of the compounds. The aspartate binding site, therefore, must be identified as that which is activated in vivo. The question of separate receptors for aspartate and glutamate then must still be resolved. / Science, Faculty of / Zoology, Department of / Graduate

Digital averaging and analysis of miniature endplate currents (MEPCs) from mouse diaphragm was used to characterize the normal MEPC and its modification by a variety of drugs. Under normal conditions the decay of MEPCs showed consistent deviations from a simple exponential consisting in a progressive increase of rate constant, followed by a slow tail. Receptor blockade by d-tubocurarine (dTC), a-bungarotoxin, and other agents thought to occupy ACh-binding sites reduced MEPC amplitude, accelerated MEPC decay by about 30% (making it about equal to decay rate of channels opened by exogenous acetylcholine), and eliminated the early deviations from an exponential decay; dTC also abolished the late tail. Examination of the interaction of acetylcholinesterase (AChE) poisoning and receptor blockade on MEPC height and time course indicated that normally most quantal ACh is captured by receptors and, as predicted by theoretical consideration, a rather large degree of receptor blockade is necessary to reduce MEPC height. MEPC tails were exaggerated by AChE poisoning and exogenous ACh or carba-chol. The latter agents reduced MEPC height in a fashion inconsistent with blockade of ACh binding and concurrent modulation of the tail suggested an important role of desensitized receptors in tail generation. A number of other drug actions are also described quantitatively: (a) channel prolongation, typical of alcohols but also found with ketones and some amines; (b) 'channel plugging', typical of local anaesthetics but also found with many other agents, including long chain alcohols, and (c) an action to reduce MEPC size without reducing net response to exogenous agonist typical of volatile anaesthetics, associated with increase rather than decrease of ACh binding to receptor. Criteria for distinguishing different modes of modification of receptor function are discussed. / Medicine, Faculty of / Anesthesiology, Pharmacology and Therapeutics, Department of / Graduate

In this dissertation I describe the isolation and characterization of genomic clones for the human synapsin I gene, the establishment of a linkage map for the human synapsin I gene locus, and studies of the possible involvement of this gene in neurological disease. Synapsin I is a neuron-specific phosphoprotein which is concentrated at the presynaptic terminal. Evidence suggests that it plays a fundamental role in the regulation of neurotransmitter release. Altogether 27,500 bp of the human synapsin I gene have been isolated, and the gene structure has been partially determined. DNA sequence comparisons between human and rat genes show a high degree of conservation. Sequenced exons display an 87% identity to each other. The synapsin I genomic clones were employed in the search for a polymorphic marker. A compound (AC)n repeat located 1000 base pairs downstream from the human synapsin I gene and within the last intron of the A-raf-1 gene has been identified. DNA database comparisons of the sequences surrounding the repeat indicate that the synapsin I gene and the A-raf-1 gene lie immediately adjacent to each other, in opposite orientation. Polymerase chain reaction amplification of this synapsin I / A-raf-1 associated repeat using total genomic DNA from members of the 40 reference pedigree families of the Centre d'Etude du Polymorphisme Humaine showed it to be highly polymorphic, with a polymorphic information content value of 0.84 and a minimum of eight alleles. Because the synapsin I gene had been mapped previously to the short arm of the human X chromosome at Xp11.2, linkage analysis was performed with markers on the proximal short arm of the X chromosome. The most likely gene order is: DXS7 - SYN/ARAF1 - TIMP - DXS255 - DXS146 with a relative probability of 5 x 108 compared with the next most likely order. The SynI/Araf marker was next utilized in a linkage study aimed at establishing a more accurate placement of the genetic locus responsible for the ocular disorder Congenital stationary night blindness, which had been mapped previously close to DXS7. Our results confirm this prior localization and also exclude any placement proximal to the SYN/ARAF1 locus. Finally, the inheritance of the different alleles of the SynI/Araf marker in three families with Rett syndrome, a severe neurodegenerative disorder, which has been assigned to the X chromosome, was studied. In at least one of the families in which two half sisters with the same mother suffer from the disease, the inheritance of Rett syndrome was discordant with the inheritance of the same allele for the SynI/Araf marker. Thus, this highly informative repeat has proven already effective in the study of X-linked diseases and should serve as a valuable marker for disease loci mapped to the Xp11 region.

Thesis (Ph.D.) -- University of Texas Southwestern Medical Center at Dallas, 2005. / Not embargoed. Vita. Bibliography: 229-249.

How does a nervous system orchestrate compound behaviors? Finding the neural basis of behavior requires knowing which neurons control the behavior and how they are connected. To accomplish this we measured and manipulated neural activity in a live, behaving animal with a completely defined connectome. The C. elegans escape response is a compound behavior consisting of a sequence of behavioral motifs. Gentle touch induces a reversal and suppression of head movements, followed by a deep turn allowing the animal to navigate away from the stimulus. The connectome provides a framework for the neural circuit that controls this behavior. We used optical physiology to determine the activity patterns of individual neurons during the behavior. Calcium imaging of locomotion interneurons and motor neurons reveal unique activity profiles during different motifs of the escape response. Furthermore, we used optogenetics and laser ablations to determine the contribution of individual neurons to each motif. We show these that the suppression of head movements and turning motifs are distinct motor programs and can be uncoupled from the reversal. The molecular mechanisms that regulate these motifs involve from signaling with the neurotransmitter tyramine. Tyramine signaling and gap junctions between locomotion interneurons and motor neurons regulate the temporal orchestration of the turning motif with the reversal. Additionally, tyramine signaling through a GPCR in GABAergic neurons facilitates the asymmetric turning during forward viii locomotion. The combination of optical tools and genetics allows us to dissect a how a neural circuit converts sensory information into a compound behavior.

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