The DHR1 domain of DOCK180 binds to SNX5 and regulates cation-independent mannose 6-phosphate receptor transport

DOCK180 is the archetype of the DOCK180-family guanine nucleotide exchange factor for small GTPases Rac1 and Cdc42. DOCK180-family proteins share two conserved domains, called DOCK homology region (DHR)-1 and -2. Although the function of DHR2 is to activate Rac1, DHR1 is required for binding to phosphoinositides. To better understand the function of DHR1, we searched for its binding partners by direct nanoflow liquid chromatography/tandem mass spectrometry, and we identified sorting nexins (SNX) 1, 2, 5, and 6, which make up a multimeric protein complex mediating endosome-to-trans-Golgi-network (TGN) retrograde transport of the cation-independent mannose 6-phosphate receptor (CI-MPR). Among these SNX proteins, SNX5 was coimmunoprecipitated with DOCK180 most efficiently. In agreement with this observation, DOCK180 colocalized with SNX5 at endosomes. The RNA interference-mediated knockdowns of SNX5 and DOCK180, but not Rac1, resulted in the redistribution of CI-MPR from TGN to endosomes. Furthermore, expression of the DOCK180 DHR1 domain was sufficient to restore the perturbed CI-MPR distribution in DOCK180 knockdown cells. These data suggest that DOCK180 regulates CI-MPR trafficking via SNX5 and that this function is independent of its guanine nucleotide exchange factor activity toward Rac1.

Salmeterol enhances the cardiac response to gene therapy in Pompe disease.

Enzyme replacement therapy (ERT) with recombinant human (rh) acid α-glucosidase (GAA) has prolonged the survival of patients. However, the paucity of cation-independent mannose-6-phosphate receptor (CI-MPR) in skeletal muscle, where it is needed to take up rhGAA, correlated with a poor response to ERT by muscle in Pompe disease. Clenbuterol, a selective β2 receptor agonist, enhanced the CI-MPR expression in striated muscle through Igf-1 mediated muscle hypertrophy, which correlated with increased CI-MPR (also the Igf-2 receptor) expression. In this study we have evaluated 4 new drugs in GAA knockout (KO) mice in combination with an adeno-associated virus (AAV) vector encoding human GAA, 3 alternative β2 agonists and dehydroepiandrosterone (DHEA). Mice were injected with AAV2/9-CBhGAA (1E+11 vector particles) at a dose that was not effective at clearing glycogen storage from the heart. Heart GAA activity was significantly increased by either salmeterol (p<0.01) or DHEA (p<0.05), in comparison with untreated mice. Furthermore, glycogen content was reduced in the heart by treatment with DHEA (p<0.001), salmeterol (p<0.05), formoterol (p<0.01), or clenbuterol (p<0.01) in combination with the AAV vector, in comparison with untreated GAA-KO mice. Wirehang testing revealed that salmeterol and the AAV vector significantly increased performance, in comparison with the AAV vector alone (p<0.001). Similarly, salmeterol with the vector increased performance significantly more than any of the other drugs. The most effective individual drugs had no significant effect in absence of vector, in comparison with untreated mice. Thus, salmeterol should be further developed as adjunctive therapy in combination with either ERT or gene therapy for Pompe disease.

Propriedades Psicométricas da Figura Complexa de Rey numa Amostra de Adultos da População Portuguesa

A Figura Complexa de Rey-Osterrieth (FCR-O) é um teste de avaliação neuropsicológica realizado em três momentos, que avalia a habilidade visuoespacial, organização percetiva, aptidão visuoespacial construtiva, memória visual, atenção, planificação e função motora. Esta investigação tem como objetivo avaliar as propriedades psicométricas da FCR-O, numa amostra de adultos da população portuguesa. O protocolo de investigação incluiu uma bateria de testes neuropsicológicos: Figura Complexa de Rey (FCR-O), Teste do Desenho do Relógio (TDR), Bateria de Avaliação Frontal (FAB), Rey 15 Item, Matrizes Progressivas de Raven (MPR) e a Escala de Auto-Avaliação da Ansiedade de Zung. A amostra deste estudo abrangeu 453 sujeitos, 192 homens e 261 mulheres, com idades compreendidas entre os 18 e os 90 anos (M = 40,44; DP = 19,78). Os resultados principais deste estudo indicam que a FCR-O é influenciada por diversas variáveis, tais como a idade, sexo, escolaridade, profissão, residência e regiões, apresentando uma validade convergente adequada, com correlações positivas com o TDR, FAB e MPR. A título conclusivo, verificámos que a FCR-O tem caraterísticas psicométricas satisfatórias, especificamente da consistência interna, concordância entre juízes e estabilidade temporal, sugerindo a sua utilização em populações não clínicas. / The Rey–Osterrieth complex figure (ROCF) is a neuropsychological test which was conducted in three phases to assess visuospatial ability, perceptive organization, constructive visuospatial ability, visual memory, attention, planning and motor function. This investigations aims to evaluate the psychometric properties of ROCF, in an adult sample of the Portuguese population. The investigation protocol includes a battery of neuropsychological tests: Complex Figure of Rey, Clock Drawing Test, Frontal Assessment Battery, Rey Item 15, Raven Progressive Matrices and the Zung Self-Rating Anxiety Scale. The sample of this study consisted of 453 subjects, 192 men and 261 women, aged between 18 and 90 years old (M = 40.44, SD = 19.78). The result of this study indicate that the ROCF is influenced by several variables, such as age, sex, education, profession, residence and regions. It also has adequate convergent validity, with positive correlations with TDR, FAB and MPR. In conclusion, we verified that the ROCF is a useful instrument to early detect some neuropsychological deficits. It revealed satisfactory psychometric characteristics, specifically in internal consistency, agreement between judges and temporal stability, suggesting its usage in no clinical populations.

Estudo Preliminar das Propriedades Psicométricas e dos Dados Normativos da Forma Geral das Matrizes Progressivas de Raven numa Amostra da Comunidade

Objetivo: Estudar as propriedades psicométricas e os dados normativos da Forma Geral das Matrizes Progressivas de Raven numa amostra da comunidade da população portuguesa. Método: A amostra é constituída por 697 pessoas (314 homens e 383 mulheres), com idades compreendidas entre os 12 e os 90 anos. Todos os participantes preencheram uma declaração de consentimento informado e uma bateria de testes neuropsicológicos, incluindo a Forma Geral das Matrizes Progressivas de Raven (FG-MPR), Teste de Memória de 15-Item de Rey, Escala de Autoavaliação de Ansiedade de Zung, Bateria de Avaliação Frontal e Figura Complexa de Rey. Resultados: A média na FG-MPR foi de 44,47 (DP = 10,78). Os resultados demonstraram que todas as variáveis sociodemográficas (idade, sexo, escolaridade, profissão, regiões e tipologia de áreas urbanas), exceto o estado civil, apresentaram ter influência significativa nas pontuações da FG-MPR. A confiabilidade e a estabilidade temporal da FG-MPR revelaram-se adequadas. A análise fatorial exploratória e confirmatória mostrou que o modelo para um fator não é adequado. Um modelo a quatro fatores continua a não ser adequado. Conclusão: Os dados do presente estudo sugerem que se trata de um instrumento com potencialidades na sua utilização junto da população portuguesa. / Purpose: To study the psychometric properties and date normative of the Raven’s Standard Progressive Matrices in a Portuguese community sample. Method: The sample consists of 697 people (314 men and 383 women), aged between 12 and 90 years. All participants filled an informed consent form and a battery of neuropsychological tests, which included Raven’s Standard Progressive Matrices (RSPM), Rey 15-Item Memory Test, Zung Self-Rating Anxiety Scale, Frontal Assessment Battery, and Rey Complex Figure Test. Results: The average in RSPM was 44.47 (SD = 10.78). The results showed that all of the sociodemographic variables (age, sex, education, profession, region, and typology of urban areas), with the exception of civil status, showed significant influence on RSPM scores. The reliability and temporal stability of RSPM were adequate. Exploratory and Confirmatory factor analysis showed that the model is not better explained by one factor. A two-factor model was not also suitable. Conclusion: The data from this study suggest that it is an instrument with potential for its use among the Portuguese population.

Random Access over Multi-Packet Reception Channels: A Mean-Field Approach

Thesis (Ph.D.)--University of Washington, 2016-06

Studies of antihypertensive drug persistence and adherence in the Glasgow Blood Pressure Clinic

Hypertension (HTN) is a major risk factor for cardiovascular diseases including stroke, coronary heart disease (CHD), chronic renal failure, peripheral vascular disease, myocardial infarction, congestive heart failure and premature death. The prevalence of HTN in Scotland is very high and although a high proportion of the patients receive antihypertensive medications, blood pressure (BP) control is very low. Recommendations for starting a specific antihypertensive class have been debated between various guidelines over the years. Some guidelines and HTN studies have preferred to start with a combination of an antihypertensive class instead of using a single therapy, and they have found greater BP reductions with combination therapies than with monotherapy. However, it has been shown in several clinical trials that 20% to 35% of hypertensive patients could not achieve the target BP, even though they received more than three antihypertensive medications. Several factors were found to affect BP control. Adherence and persistence were considered as the factors contributing the most to uncontrolled hypertension. Other factors such as age, sex, body mass index (BMI), alcohol intake, baseline systolic BP (SBP), and the communication between physicians and patients have been shown to be associated with uncontrolled BP and resistant hypertension. Persistence, adherence and compliance are interchangeable terms and have been used in the literature to describe a patient’s behaviour with their antihypertensive drugs and prescriptions. The methods used to determine persistence and adherence, as well as the inclusion and exclusion criteria, vary between persistence and adherence studies. The prevalence of persistence and adherence have varied between these studies, and were determined to be high in some studies and low in others. The initiation of a specific antihypertensive class has frequently been associated with an increase or decrease in adherence and persistence. The tolerability and efficacy of the initial antihypertensive class have been the most common methods of explaining this association. There are also many factors that suggest a relationship with adherence and persistence. Some factors in previous studies, such as age, were frequently associated with adherence and persistence. On the other hand, relationships with certain factors have varied between the studies. The associations of age, sex, alcohol use, smoking, baseline systolic blood pressure (SBP) and diastolic BP (DBP), the presence of comorbidities, an increase in the number of pills and the relationship between patients and physicians with adherence and persistence have been the most commonly investigated factors. Most studies have defined persistence in terms of a patient still taking medication after a period of time. A medication possession ratio (MPR) ≥ 80 has been used to define compliance. Either of these terminologies, or both, have been used to estimate adherence. In this study, I used the same definition for persistence to identify patients who have continued with their initial treatment, and used persistence and MPR to define patients who adhered to their initial treatment. The aim of this study was to estimate the prevalence of persistence and adherence in Scotland. Also, factors that could have had an effect on persistence and adherence were studied. The number of antihypertensive drugs taken by patients during the study and factors that led to an increase in patients being on a combination therapy were also evaluated. The prevalence of resistance and BP control were determined by taking the BP after the last drug had been taken by persistent patients during five follow-up studies. The relationship of factors such as age, sex, BMI, alcohol use, smoking, estimated glomerular filtration rate (eGFR), and albumin levels with BP reductions for each antihypertensive class were determined. Information Services Division (ISD) data, which includes all antihypertensive drugs, were collected from pharmacies in Scotland and linked to the Glasgow Blood Pressure Clinic (GBPC) database. This database also includes demographic characteristics, BP readings and clinical results for all patients attending the GBPC. The case notes for patients who attended the GBPC were reviewed and all new antihypertensive drugs that were prescribed between visits, BP before and after taking drugs, and any changes in the hypertensive drugs were recorded. A total of 4,232 hypertensive patients were included in the first study. The first study showed that angiotensin converting enzyme inhibitor (ACEI) and beta-blockers (BB) were the most prescribed antihypertensive classes between 2004 and 2013. Calcium channel blockers (CCB), thiazide diuretics and angiotensin receptor blockers (ARB) followed ACEI and BB as the most prescribed drugs during the same period. The prescription trend of the antihypertensive class has changed over the years with an increase in prescriptions for ACEI and ARB and a decrease in prescriptions for BB and diuretics. I observed a difference in antihypertensive class prescriptions by age, sex, SBP and BMI. For example, CCB, thiazide diuretics and alpha-blockers were more likely to be prescribed to older patients, while ACEI, ARB or BB were more commonly prescribed for younger patients. In a second study, 4,232 and 3,149 hypertensive patients were included to investigate the prevalence of persistence in the Scottish population in 1- and 5-year studies, respectively. The prevalence of persistence in the 1-year study was 72.9%, while it was only 62.8% in the 5-year study. Those patients taking ARB and ACEI showed high rates of persistence and those taking diuretics and alpha blockers had low rates of persistence. The association of persistence with clinical characteristics was also investigated. Younger patients were more likely to totally stop their treatment before restarting their treatment with other antihypertensive drugs. Furthermore, patients who had high SBP tended to be non-persistent. In a third study, 3,085 and 1,979 patients who persisted with their treatment were included. In the first part of the study, MPR was calculated, and patients with an MPR ≥ 80 were considered as adherent. Adherence rates were 29.9% and 23.4% in the 1- and 5-year studies, respectively. Patients who initiated the study with ACEI were more likely to adhere to their treatments. However, patients who initiated the study with thiazide diuretics were less likely to adhere to their treatments. Sex, age and BMI were different between the adherence and non-adherence groups. Age was an independent factor affecting adherence rates during both the 1- and 5-year studies with older patients being more likely to be adherent. In the second part of the study, pharmacy databases were checked with patients' case notes to compare antihypertensive drugs that were collected from the pharmacy with the antihypertensive prescription given during the patient’s clinical visit. While 78.6% of the antihypertensive drugs were collected between clinical visits, 21.4% were not collected. Patients who had more days to see the doctor in the subsequent visit were more likely to not collect their prescriptions. In a fourth study, 3,085 and 1,979 persistent patients were included to calculate the number of antihypertensive classes that were added to the initial drug during the 1-year and 5-year studies, respectively. Patients who continued with treatment as a monotherapy and who needed a combination therapy were investigated during the 1- and 5-year studies. In all, 55.8% used antihypertensive drugs as a monotherapy and 44.2% used them as a combination therapy during the 1-year study. While 28.2% of patients continued with treatment without the required additional therapy, 71.8% of the patients needed additional therapy. In all, 20.8% and 46.5% of patients required three different antihypertensive classes or more during the 1-year and 5-year studies, respectively. Patients who started with ACEI, ARB and BB were more likely to continue as monotherapy and less likely to need two more antihypertensive drugs compared with those who started with alpha-blockers, non-thiazide diuretics and CCB. Older ages, high BMI levels, high SBP and high alcohol intake were independent factors that led to an increase in the probability of patients taking combination therapies. In the first part of the final study, BPs were recorded after the last drug had been taken during the 5 year study. There were 815 persistent patients who were assigned for this purpose. Of these, 39% had taken one, two or three antihypertensive classes and had controlled BP (controlled hypertension [HTN]), 29% of them took one or two antihypertensive classes and had uncontrolled BP (uncontrolled HTN), and 32% of the patients took three antihypertensive classes or more and had uncontrolled BP (resistant HTN). The initiation of an antihypertensive drug and the factors affecting BP pressure were compared between the resistant and controlled HTN groups. Patients who initiated the study with ACEI were less likely to be resistant compared with those who started with alpha blockers and non-thiazide diuretics. Older patients, and high BMI tended to result in resistant HTN. In the second part of study, BP responses for patients who initiated the study with ACEI, ARB, BB, CCB and thiazide diuretics were compared. After adjusting for risk factors, patients who initiated the study with ACEI and ARB were more respondent than those who took CCB and thiazide diuretics. In the last part of this study, the association between BP reductions and factors affecting BP were tested for each antihypertensive drug. Older patients responded better to alpha blockers. Younger patients responded better to ACEI and ARB. An increase in BMI led to a decreased reduction in patients on ACEI and diuretics (thiazide and non-thiazide). An increase in albumin levels and a decrease in eGFR led to decreases in BP reductions in patients on thiazide diuretics. An increase in eGFR decreased the BP response with ACEI. In conclusion, although a high percentage of hypertensive patients in Scotland persisted with their initial drug prescription, low adherence rates were found with these patients. Approximately half of these patients required three different antihypertensive classes during the 5 years, and 32% of them had resistant HTN. Although this study was observational in nature, the large sample size in this study represented a real HTN population, and the large pharmacy data represented a real antihypertensive population, which were collected through the support of prescription data from the GBPC database. My findings suggest that ACEI, ARB and BB are less likely to require additional therapy. However, ACEI and ARB were better tolerated than BB in that they were more likely to be persistent than BB. In addition, users of ACEI, and ARB have good BP response and low resistant HTN. Linkage patients who participated in these studies with their morbidity and mortality will provide valuable information concerning the effect of adherence on morbidity and mortality and the potential benefits of using ACEI or ARB over other drugs.

El ejercicio y el desempeño musical: doce semanas de actividad física mejoran la capacidad de ejecución de un instrumento musical en estudiantes de conservatorio

Con el fin de evaluar los cambios que produce un programa de actividad física en la percepción que tienen los músicos sobre su capacidad de ejecución de un instrumento musical, se realizó una intervención con un programa de actividad física basado en la técnica Pilates, durante 12 semanas, en el Conservatorio de Música de la Universidad Nacional de Colombia, con estudiantes del programa de música instrumental. Se midieron parámetros de la aptitud física (capacidad cardiorrespiratoria, fuerza, flexibilidad y composición corporal) y de la percepción de la capacidad de ejecución (fatiga muscular, nivel de esfuerzo, dolor y fluidez) antes y después de la intervención. Los resultados arrojaron cambios positivos en la aptitud física logrando un aumento significativo en la flexibilidad y resistencia de los miembros inferiores en 14 participantes (70% de la muestra), y en la percepción de la capacidad de ejecución instrumental con el retraso en la aparición de la fatiga muscular mientras se está ejecutando el instrumento (30 minutos en promedio). Esto permite a los músicos abordar un repertorio extenso con menor fatiga, minimizando el riesgo de lesión o alteraciones musculo-esqueléticas que influyan directamente en su desempeño técnico y artístico.

Supervivencia y mortalidad por tipo de terapia de reemplazo renal, en pacientes con enfermedad renal crónica terminal, en una cohorte de pacientes asegurados en Colombia

En Colombia se ha podido establecer que la incidencia y mortalidad de la Enfermedad Renal Crónica Terminal continúan en aumento en los últimos 6 años a pesar de las estrategias de intervención para prevención y control de la enfermedad implementadas nivel nacional. Este trabajo busca establecer la línea de base para la población asegurada en Colombia, frente a la supervivencia de pacientes en terapia de remplazo renal (TRR).