ABSTRACT: UTILIZING NOVEL DOSE EQUIVALENCE METHODOLOGIES TO EXAMINE COCAINE'S EFFECTS ON THE VASCULATURE Neil S. Lamarre Doctor of Philosophy Temple University School of Medicine, 2013 Doctoral Advisory Committee Chair: Ronald J. Tallarida, Ph.D. Cocaine abuse and addiction is a serious health problem, resulting in thousands of emergency room visits and deaths each year in the United States. It is particularly toxic to the cardiovascular system, including deleterious effects on the peripheral vasculature. These effects are not well understood, but evidence suggests chronic cocaine use may lead to endothelial dysfunction, thereby increasing relative risk of a number of other cardiovascular diseases including stroke, aneurysm, myocardial infarction, hypertension, etc. Data from our lab, and others, suggest that the presence of a functional endothelium has a dramatic effect on the contractility of the rat aorta that is agonist-specific. Attenuation of this endothelium-dependent vasodilatory component of agonist action is a primary feature of endothelial dysfunction. We have utilized dose equivalence theory to calculate the dose response relationship for the endothelium-dependent vasodilatory component of an agonist causing overt vasoconstriction. This component cannot be measured directly, but our novel methodology allows us to quantitate agonist-specific impairment of vasodilation, and describe it using the familiar parameters of the dose response curve. Another strength of this method, relative to currently used in vitro methods, is that it also avoids the confounding variable of a second agonist used to produce the initial vasoconstriction. To validate the methodology, a pilot study was performed examining the endothelial dysfunction in STZ-induced diabetic rats, as a positive control for endothelial dysfunction. Interestingly, this treatment showed impairment in the endothelium-dependent vasodilatory component of action of norepinephrine, but not of angiotensin-II. Thus, our initial hypothesis was confirmed - that disruption of the vasodilatory components of various agonists are independent, and that agonist-specific information may prove useful. Next, we employed our new methodology utilizing the rat aorta as our vascular model to test the hypothesis that chronic cocaine administration causes endothelial dysfunction. We first examined the endothelium-dependent vasodilation component of a number of physiologically important vasoconstrictors, and attempted to determine which vasodilatory mediators contributed to the effect. We found the endothelium to have a profound effect on the dose response curve to three important endogenous agonists. These data suggest that under conditions of endothelial dysfunction exaggerated vasoconstriction could occur, even within normal plasma concentration ranges of these vasoconstrictors, resulting in elevated blood pressure and further damage to the endothelium over time. No endothelial dysfunction was observed with this treatment paradigm, using our methodology or the standard approach. This may be a result of insufficient duration of cocaine treatment, or a result of our selection of the rat aorta as a model. We wanted to further investigate which vasodilatory mechanisms were involved in this vasodilatory component of action. We inhibiting various endothelium-derived mediators of this vasodilatory component of action (such as nitric oxide or prostacyclin), which revealed differential activation of these mediators by the agonists examined. For example, inhibition of nitric oxide synthesis abolished the endothelium-dependent vasodilatory component of endothelin-1, but only partially attenuated that of angiotensin-II. Thus, the agonist-specific pattern of impairment may also prove useful in examining the underlying mechanisms of impaired vasodilation. Endothelial dysfunction is one reported consequence of long term cocaine abuse; however, there are conflicting reports on the acute vascular effects of cocaine, with some reports concluding that cocaine is a vasoconstrictor, and some reporting its action as a vasodilator. There are in vitro reports of cocaine causing release of vasoconstrictors from the endothelium, which supports the longstanding notion of cocaine as a vasoconstrictor. However, one recent report demonstrates a dose-dependent vasodilatory effect of cocaine in rat aorta that is independent of the endothelium. This complexity is perhaps due, in part, to cocaine's affinity for a number of molecular targets, acting in combination. In examining the acute action of cocaine in our preparation, we observed an "inverted-U" shaped dose response, also referred to as a hormetic dose response curve. We then applied dose equivalence methodology in order to derive the "unknown" second component contributing the vasodilatory action of cocaine at higher doses. This methodology lets us calculate this unknown component, and describe it with the familiar parameters of a dose response curve, which could potentially aid in the identification of the unknown component. The preliminary studies with acute cocaine utilized a sub-maximal dose of phenylephrine in order to observe tension changes in either direction. This prompted us to further characterize the interaction of cocaine with other alpha adrenoceptor agonists. Importantly, because cocaine alone had no effect at doses up to 100 µM, but potentiated the vasoconstriction of alpha agonists, the interaction is therefore synergistic. This constitutes evidence of a previously undescribed mechanism contributing to cocaine's vasoconstricting effect. In vivo, reuptake inhibition is a major mechanism for cocaine-induced vasoconstriction, but is excluded in this experiment by virtue of low levels of sympathetic innervation in the rat aorta, and the use of methoxamine, an alpha agonist not subject to the reuptake mechanisms. This interaction may contribute to cocaine-induced vasoconstriction in the coronary arteries, especially in circumstances of endothelial dysfunction. In summary, the work presented in this dissertation applies new methodologies utilizing dose equivalence theory to the study of cocaine's effects on peripheral vasculature, and presents novel findings of synergy with respect to cocaine's enhancement on the action of alpha adrenoceptor-mediated vasoconstriction. / Pharmacology

The Hulver laboratory has recently found that the neuropeptide Y Y1 receptor (NPY Y1R) mRNA expression is elevated in skeletal muscle of obese humans (Hulver, unpublished). The goal of this research is to study the role of the NPY Y1R in skeletal muscle lipid metabolism. Rat L6, mouse C2C12, and human primary myotubes were incubated in 14C palmitate labeled fatty acid oxidation medium containing 80ng/mL, 250ng/mL, and 500ng/mL of NPY and for a three hour period. Experiments were repeated with the addition of 17mg/mL diprotin A to each NPY treatment. Fatty acid oxidation (FAO) and the percentage of lipids stored within the myotubes as diacylglyceride (DAG) and triaclyglyceride (TAG) were measured. Analyses were repeated in rat L6 and mouse C2C12 following a three hour incubation in 14C palmitate labeled fatty acid oxidation medium containing 1µg/mL, 10µg/mL, and 50µg/mL of the NPY Y1R ligand, [Leu31, Pro34] neuropeptide Y (Bachem, Torrance, CA). Incubation of human primary myotubes in NPY treatments with the addition of diprotin A significantly increased TAG accumulation (p< 0.05). Mouse C2C12 mytoube incubation in 500ng/mL NPY with diprotin A increased FAO (p 0.05). All other NPY and NPY Y1R ligand treatments in had no significant effect on FAO or the accumulation of TAG and DAG. / Master of Science

Yes / Background: To determine the test-retest reproducibility of accommodative facility (AF) measures in an unselected sample of UK primary school children. Methods: Using 2.00 DS flippers and a viewing distance of 40 cm, AF was measured in 136 children (range 4–12 years, average 8.1 2.1) by five testers on three occasions (average interval between successive tests: eight days, range 1–21 days). On each occasion, AF was measured monocularly and binocularly, for two minutes. Full datasets were obtained in 111 children (81.6 per cent). Results: Intra-individual variation in AF was large (standard deviation [SD] = 3.8 cycles per minute [cpm]) and there was variation due to the identity of the tester (SD = 1.6 cpm). On average, AF was greater: (i) in monocular compared to binocular testing (by 1.4 cpm, p < 0.001); (ii) in the second minute of testing compared to the first (by 1.3 cpm, p < 0.001);(iii) in older compared to younger children (for example, AF for 4/5-year-olds was 3.3 cpm lower than in children ≥10 years old, p = 0.009); and (iv) on subsequent testing occasions (for example, visit-2 AF was 2.0 cpm higher than visit-1 AF, p < 0.001). After the first minute of testing at visit-1, only 36.9 per cent of children exceeded published normative values for AF (≥11 cpm monocularly and≥8 cpm binocularly), but this rose to 83.8 per cent after the third test. Using less stringent pass criteria (≥6 cpm monocularly and≥3 cpm binocularly), the equivalent figures were 82.9 and 96.4 per cent, respectively. Reduced AF did not co-exist with abnormal near point of accommodation or reduced visual acuity. Conclusions: The results reveal considerable intra-individual variability in raw AF measures in children. When the results are considered as pass/fail, children who initially exhibit normal AF continued to do so on repeat testing. Conversely, the vast majority of children with initially reduced AF exhibit normal performance on repeat testing. Using established pass/fail criteria, the prevalence of persistently reduced AF in this sample is 3.6 per cent. / UK College of Optometrists

Made available in DSpace on 2017-07-10T14:17:10Z (GMT). No. of bitstreams: 1 STEFANY_ PEDROTTI 2015.pdf: 2292343 bytes, checksum: 2d00d0ff8c02256c8040bdbd293e6156 (MD5) Previous issue date: 2015-02-27 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Periodontal disease is characterized by a chronic infection, which may cause systemic inflammation due to high levels of inflammatory cytokines such as tumor necrosis factor-alpha (TNF-&#945;), interleukin-6 (IL-6), interleukin-8 (IL-8) and interleukin-18 (IL-18), contributing to the development of endothelial dysfunction in various organs. The impairment of endothelial function can lead to erectile dysfunction, because it is characterized by vascular changes such as occlusion of cavernous arteries by atherosclerosis and reduction vascular reactivity due to decreased release of nitric oxide. In this sense, inflammation associated with periodontal disease may damage the penile endothelial layer, leading to occurrence of erectile dysfunctional. However, the potential role of chronic periodontitis in the pathophysiology of erectile dysfunction remains scarce. Therefore, the aim of the study was to evaluate the tissue morphology of periodontal and penis rats associated or not with experimentally induced periodontitis. For this, 18 Wistar rats were obtained and induced periodontal disease in 9 rats, whereas the remaining 9 rats served as controls. After 30 days of induction of periodontal disease, the sexual behavior of rats was recorded and later, the animals were sacrificed to obtain gingival sample, hemimandible and penis to the morphological analysis, morphometric, immunological and radiographic. The results showed that experimental periodontitis contributed to the reduction in the sexual behavior of rats (p<0.05) and caused alveolar bone loss with a significant increase in osteoclastic activity and gingival volume (p<0.05). Moreover, the group with periodontitis had a higher concentration of inflammatory cytokines such as IL-6 in the gingiva and TNF-&#945; in the penis (p<0.05). There was no statistically significant difference in ventral-dorsal and side-to-side distance of the penis (p> 0.05), however there was a higher thickness and a shorter area statistically significant of penile dorsal artery in the group with experimental periodontitis, as well as a smaller area of vascular spaces of the penile cavernous tissue (p<0.05). Based on these results, it is concluded that the experimental periodontitis caused structural and functional changes in the penis Wistar rats and sexual behavior changes, therefore systemic inflammation caused by periodontal disease can be an important risk factor for erectile dysfunction, that is why is necessary the development of interventions involving many health professionals for prevention and treatment of these two diseases that endanger the health and general well-being of individuals / A doença periodontal caracteriza-se por ser uma infecção crônica, a qual pode causar uma inflamação sistêmica devido aos elevados níveis de citocinas inflamatórias envolvidas, tal como fator de necrose tumoral-alfa (TNF-&#945;), interleucina-6 (IL-6), interleucina-8 (IL-8) e interleucina-18 (IL-18), o que contribui para o desenvolvimento da disfunção endotelial em diferentes órgãos. O comprometimento da função endotelial pode levar à disfunção erétil, pois a mesma é caracterizada por alterações vasculares, como oclusão das artérias cavernosas por aterosclerose e redução da reatividade vascular devido à diminuição da liberação do óxido nítrico. Nesse sentido, a inflamação associada com a doença periodontal pode danificar as camadas endoteliais penianas, induzindo à ocorrência da ereção disfuncional. Contudo, o papel potencial da periodontite crônica na fisiopatologia da disfunção erétil permanece escasso. Portanto, o objetivo deste estudo foi avaliar a morfologia tecidual do periodonto e pênis de ratos associados ou não com a periodontite induzida experimentalmente. Para isso, foram obtidos 18 ratos Wistar e induzida a doença periodontal em 9 ratos, sendo que os 9 ratos restantes serviram como controle. Aos 30 dias de indução da doença periodontal, foi gravado o comportamento sexual dos ratos e após, os animais foram sacrificados para a obtenção de amostra gengival, hemimandíbula e pênis para as análises morfológicas, morfométricas, imunológicas e radiográficas. Os resultados mostraram que a periodontite experimental contribuiu para a redução do comportamento sexual dos ratos (p<0.05), bem como causou perda óssea alveolar, com significante atividade osteoclástica e aumento de volume gengival (p<0.05). Além disso, o grupo com periodontite apresentou uma maior concentração de citocinas inflamatórias, tal como IL-6 nas gengivas e TNF-&#945; no pênis (p<0.05). Não houve diferença estatisticamente significativa nas dimensões ventral-dorsal e látero-lateral do pênis (p>0.05), no entanto verificou-se uma maior espessura e menor área estatisticamente significante da artéria dorsal do pênis no grupo com periodontite experimental, assim como uma menor área dos espaços vasculares dos corpos cavernosos (p<0.05). Com base nesses resultados, conclui-se que a peridontite experimental provocou alterações estruturais e funcionais no pênis de ratos Wistar, bem como alterações de comportamento sexual, portanto a inflamação sistêmica causada pela doença periodontal pode ser um importante fator de risco para a disfunção erétil, por isso se faz necessário o desenvolvimento de intervenções envolvendo vários profissionais da saúde para prevenção e tratamento dessas duas patologias que comprometem a saúde e bem-estar geral dos indivíduos

Background and objective: Major risk factors for congestive heart failure (CHF) are myocardial infarction, hypertension, diabetes, atrial fibrillation, smoking, left ventricular hypertrophy (LVH) and obesity. However, since these risk factors only explain part of the risk of CHF, we investigated whether persistent organic pollutants (POPs) might also play a role. Methods: In the Prospective Investigation of the Vasculature in Uppsala Seniors (PIVUS) study, left ventricular ejection fraction, (EF), E/A-ratio and isovolumic relaxation time (IVRT), were determined by echocardiography and serum samples of 21 POPs were analyzed in serum measured by high-resolution chromatography coupled to high-resolution mass spectrometry (HRGC/HRMS) in 998 subjects all aged 70 years. Results: In this cross-sectional analysis, high levels of several of the polychlorinated biphenyls (PCB congeners 99, 118, 105, 138, 153, and 180) and octachlorodibenzo-p-dioxin (OCDD) were significantly related to a decreased EF. Some POPs were also related to a decreased E/A-ratio (PCBs 206 and 209). All the results were adjusted for gender, hypertension, diabetes, smoking, LVH and BMI, and subjects with myocardial infarction or atrial fibrillation were excluded from the analysis. Conclusions: Circulating levels of POPs were related to impairments in both left ventricular systolic and diastolic function independently of major congestive heart failure risk factors, suggesting a possible role of POPs in heart failure.

名古屋大学博士学位論文 学位の種類 : 博士(医学)(論文) 学位授与年月日:平成5年2月19日 竹中晃氏の博士論文として提出された

Thesis (M.S.)--University of Michigan, 1988.

Thesis (doctoral)--Karolinska Institutet, Stockholm, 1990. / Extra t.p. with thesis statement inserted. Includes bibliographical references.

Made available in DSpace on 2015-07-22T20:49:11Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-27. Added 1 bitstream(s) on 2015-08-11T03:25:33Z : No. of bitstreams: 1 Publico-008.pdf: 1147299 bytes, checksum: c53fe718c40d4278be6155456942dfbe (MD5) / Contexto: a disfunção sexual é frequente entre pacientes com esquizofrenia, sendo relatada como um dos mais incômodos efeitos adversos dos antipsicóticos e esta diretamente relacionada com adesão ao tratamento. Objetivo: a) avaliar a frequência da disfunção sexual em uma amostra de pacientes do espectro da esquizofrenia em tratamento com antipsicóticos; b) investigar 0 efeito dos diferentes antipsicóticos na função sexual; e c) avaliar a acurácia da Escala de Experiência Sexual do Arizona (AS EX) para identificar disfunção sexual. Método: pacientes ambulatoriais com esquizofrenia ou transtorno esquizoafetivo foram entrevistados através de questionários: ASEX e Escala Dickson-Glazer (DGSFi) para avaliação do funcionamento sexual, em uma única entrevista. Resultados: 137 pacientes foram entrevistados. A sensibilidade e especificidade da ASEX em relação a DGSFi foram: 80.8% ( 95% IC= 70.0%-88.5%) e 88.1 % (95% IC=76.5%-94.7%), e a taxa de classificação incorreta foi 9.5%. A curva ROC comparando a pontuação da ASEX e DGSFi revelou valor de 0.93 (IC=0.879¬0.970) com 0 ponto de corte da ASEX encontrando sendo 14/15. A disfunção sexual foi mais alta entre as mulheres (79.2%) do que nos homens (33.3%) (X2=27.41, gl=1, p<0.001). Conclusão: pacientes em tratamento com antipsicóticos mostraram alta frequência de queixas sexuais e ASEX provou ser um instrumento eficaz para identificar disfunção sexual em amostra de pacientes ambulatoriais do espectro da esquizofrenia. Mulheres mostraram frequência mais alta de disfunção, e desejo sexual e habilidade para alcançar orgasmo foram áreas mais afetadas. 0 uso de antipsicóticos, principal mente 0 uso de combinações, foi associado com piora do funcionamento sexual.. / Background: sexual dysfunction is frequent in patients with schizophrenia, it is reported as one of the most distressing antipsychotic’s adverse effects and it is directly related to treatment compliance. Objective: a) to assess the frequency of sexual dysfunction in a sample of outpatients with schizophrenia and schizoaffective disorder under antipsychotic therapy; b) to investigate the effect of different antipsychotics on sexual function; and c) to evaluate the accuracy of the Arizona Sexual Experience Scale (ASEX) to identify sexual dysfunction. Method: Outpatients with schizophrenia or schizoaffective disorder were asked to fulfill both the ASEX and the Dickson Glazer Scale for the Assessment of Sexual Functioning Inventory (DGSFi) at a single interview. Results: 137 patients were interwied. The sensitivity and specificity of the ASEX in relation to DGSFi were: 80.8%, (95% CI= 70.0%-88.5%) and 88.1% (95% CI= 76.5%-94.7%), and the misclassification rate was 9.5%. The ROC curve comparing the ASEX and the DGSFi scores revealed a value of 0.93 (CI= 0.879-0.970), with the optimum cut-off point of ASEX being 14/15. Sexual dysfunction measured was higher in females (79.2%) than in males (33.3%) (2 = 27.41, d.f.=1, p<0.001). Discussion: Patients under antipsychotic treatment showed a high level of sexual complaints, and the ASEX proved to be an accurate instrument to identify sexual dysfunction in an outpatient sample of patients with schizophrenia spectrum. Females showed a higher frequency of sexual dysfunctions and sexual drive and ability to reach orgasm were the most affected areas. The use of antipsychotics, especially the combinations, was more likely to impair sexual functioning. / TEDE / BV UNIFESP: Teses e dissertações

Alzheimer’s disease (AD) is a neurodegenerative disorder characterized by two major pathological hallmarks, amyloid plaques and neurofibrillary tangles. The accumulation of amyloid-β protein (Aβ) is an early event associated with synaptic and mitochondrial damage in AD. Therefore, molecular pathways underlying the neurotoxicity and generation of Aβ represent promising therapeutic targets for AD. Recent studies have shown that actin severing protein, Cofilin plays an important role in synaptic remodeling, mitochondrial dysfunction, and AD pathogenesis. However, whether Cofilin is an essential component of AD pathogenesis and how Aβ induced neurotoxicity impinges its signals to Cofilin are unclear. In my dissertation studies, we found Aβ oligomers bind with intermediate activation conformers of β1-integrin to induce the loss of surface β1-integrin and activation of Cofilin via Slingshot homology-1 (SSH1) activation. Specifically, conditional loss of β1-integrin prevented Aβ induced Cofilin activation, and allosteric modulation or activation of β1-integrin significantly reduced Aβ binding to neurons and mitigated Aβ42-induced reactive oxygen species (ROS) generation, mitochondrial dysfunction, synaptic proteins depletion, and apoptosis. Furthermore, we found that SSH1 reduction, which mitigated Cofilin activation, prevented Aβ-induced mitochondrial Cofilin translocation and apoptosis, while AD brain mitochondria contained significantly increased activated/oxidized Cofilin. In mechanistic support in vivo, we demonstrated that APP transgenic mice brains contain decreased SSH/Cofilin and SSH1/14-3-3 complexes which indicates that SSH-Cofilin activation occurred by releasing of SSH from 14-3-3. We also showed that genetic reduction in Cofilin rescues APP/Aβ-induced synaptic protein loss and gliosis, as well as impairments in synaptic plasticity and contextual memory in vivo. Our lab previously found that overexpression of the scaffolding protein RanBP9 increases Aβ production in cell lines and in transgenic mice, while promoting Cofilin activation and mitochondrial dysfunction. However, how endogenous RanBP9 activates cofilin and whether endogenous RanBP9 accelerates Aβ-induced deficits in synaptic plasticity, cofilin-dependent pathology, and cognitive impairments were unknown. In my dissertation studies, we found that endogenous RanBP9 positively regulates SSH1 levels and mediates A-induced translocation of Cofilin to mitochondria. Moreover, we demonstrated that endogenous RanBP9 mediates A-induced formation of Cofilin-actin rods in primary neurons. Endogenous level of RanBP9 was also required for Aβ-induced collapse of growth cones in immature neurons and depletion of synaptic proteins in mature neurons. In vivo, we also found APP transgenic mice exhibit significantly increased endogenous RanBP9 levels and that genetic reduction in RanBP9 rescued APP/Aβ-induced synaptic protein loss, gliosis, synaptic plasticity impairments, and contextual memory deficits. These findings indicated that endogenous RanBP9 not only promotes Aβ production but also meditate Aβ induced neurotoxicity via positively regulating SSH1. Taken together, these novel findings implicate essential involvement of β1-integrin–SSH1/RanBP9–Cofilin pathway in mitochondrial and synaptic dysfunction in AD pathogenesis.