Highly accurate RNA and DNA sequencing: Application to longstanding questions in aging and cancer

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

Physical activity, exercise, and inflammatory markers in older adults: Findings from the health, aging and body composition study

OBJECTIVES: To examine the association between physical activity and inflammatory markers, with consideration for body fatness and antioxidant use. DESIGN: Cross-sectional study, using baseline data from the Health, Aging and Body Composition Study. SETTING: Metropolitan areas surrounding Pittsburgh, Pennsylvania, and Memphis, Tennessee. PARTICIPANTS: Black and white, well-functioning men and women (N=3,075), aged 70 to 79. MEASUREMENTS: Interviewer-administered questionnaires of previous-week household, walking, exercise, and occupational/volunteer physical activities. Analysis of covariance was used to examine the association between activity level and serum C-reactive protein (CRP), interleukin-6 (IL-6), and plasma tumor necrosis factor alpha (TNFalpha) with covariate adjustment. Antioxidant supplement use (multivitamin, vitamins E or C, beta carotene) was evaluated as an effect modifier of the association. RESULTS: Higher levels of exercise were associated with lower levels of CRP (P

Hearing sensitivity and bone mineral density in older adults: the Health, Aging and Body Composition Study

Bone mineral density (BMD) may be associated with hearing loss in older adults. Demineralization of the cochlear capsule has been associated with hearing loss in those with Paget's disease of the bone and otosclerosis. Osteoporosis may also result in cochlear capsule demineralization. We hypothesized that lower hip BMD and lower heel ultrasound measurements would be associated with hearing loss in a population-based sample of 2,089 older black and white men and women. Bone parameters and hearing function were measured at the fourth clinical follow-up visit. Audiometric threshold testing was used to measure air- and bone-conduction hearing sensitivity. BMD of the hip and its subregions was measured using dual-energy X-ray absorptiometry. Calcaneal bone measurements [broadband ultrasound attenuation (BUA), speed of sound (SOS) and the quantitative ultrasound index (QUI)] were obtained using heel ultrasound. After adjusting for known hearing loss risk factors, no association was found between hearing and any of the bone measurements in whites and black women. In black men, however, lower hip BMD was associated with higher odds of hearing loss; for each standard deviation decrease in total hip BMD, the odds of hearing loss were 1.41 (95% confidence interval 1.08, 1.83), 1.39 (95% CI 1.07, 1.82) for femoral neck BMD and 1.65 (95% CI 1.26, 2.16) for trochanter BMD. Conductive hearing loss was associated with lower heel ultrasound measurements, though only among white men. The results of this study are mixed and inconclusive. Lower BMD of the hip and its subregions was associated with hearing loss among black men, but not among whites or black women. Lower measurements on heel ultrasound were associated with conductive hearing loss, though only among white men. These results suggest that axial and appendicular bone parameters may be modestly associated with hearing loss in older men, but not in women.

Volumetric and areal bone mineral density measures are associated with cardiovascular disease in older men and women: The health, aging, and body composition study

The associations of volumetric (vBMD) and areal (aBMD) bone mineral density measures with prevalent cardiovascular disease (CVD) and subclinical peripheral arterial disease (PAD) were investigated in a cohort of older men and women enrolled in the Health, Aging, and Body Composition Study. Participants were 3,075 well-functioning white and black men and women (42% black, 51% women), aged 68-80 years. Total hip, femoral neck, and trochanter aBMD were measured using dual-energy X-ray absorptiometry. Quantitative computed tomography was used to evaluate spine trabecular, integral, and cortical vBMD measures in a subgroup (n = 1,489). Logistic regression was performed to examine associations of BMD measures with CVD and PAD. The prevalence of CVD (defined by coronary heart disease, PAD, cerebrovascular disease, or congestive heart failure) was 29.8%. Among participants without CVD, 10% had subclinical PAD (defined as ankle-arm index < 0.9). Spine vBMD measures were inversely associated with CVD in men (odds ratio of integral [ORintegral] = 1.34, 95% confidence interval [CI] 1.10-1.63; ORtrabecular = 1.25, 95% CI 1.02-1.53; ORcortical = 1.36, 95% CI 1.11-1.65). In women, for each standard deviation decrease in integral vBMD, cortical vBMD, or trochanter aBMD, the odds of CVD were significantly increased by 28%, 27%, and 22%, respectively. Total hip aBMD was associated with subclinical PAD in men (OR = 1.39, 95% CI 1.03-1.84) but not in women. All associations were independent of age and shared risk factors between BMD and CVD and were not influenced by inflammatory cytokines (interleukin-6 and tumor necrosis factors-alpha). In conclusion, our results provide further evidence for an inverse association between BMD and CVD in men and women. Future research should investigate common pathophysiological links for osteoporosis and CVD.

The interface between Alzheimer's disease, normal aging, and related disorders

Since the earliest descriptions of Alzheimer's disease (AD), the presence of senile plaques (SP) and neurofibrillary tangles (NFT) have been regarded as the typical pathological hallmarks of the disease. Studies over the last twenty years, however, have reported a considerable degree of heterogeneity within the AD phenotype and as a consequence, an overlap between the pathological features of AD not only with normal aging, but also with disorders related to AD. This review discusses: 1) the degree of heterogeneity within AD, 2) the concept of an 'interface' between disorders, 3) the nature and degree of the interface between AD and normal aging, vascular dementia (VD), the tauopathies, synucleinopathies, and prion disease, and 4) whether the original status of AD should be retained or whether AD, normal aging, and the related disorders should be regarded as representing a 'continuum' of neuropathological change.

Artificial intelligence and criminal evidence: A few topics

Abstract not available

Telomeres in aging and disease: lessons from zebrafish

Age is the highest risk factor for some of the most prevalent human diseases, including cancer. Telomere shortening is thought to play a central role in the aging process in humans. The link between telomeres and aging is highlighted by the fact that genetic diseases causing telomerase deficiency are associated with premature aging and increased risk of cancer. For the last two decades, this link has been mostly investigated using mice that have long telomeres. However, zebrafish has recently emerged as a powerful and complementary model system to study telomere biology. Zebrafish possess human-like short telomeres that progressively decline with age, reaching lengths in old age that are observed when telomerase is mutated. The extensive characterization of its well-conserved molecular and cellular physiology makes this vertebrate an excellent model to unravel the underlying relationship between telomere shortening, tissue regeneration, aging and disease. In this Review, we explore the advantages of using zebrafish in telomere research and discuss the primary discoveries made in this model that have contributed to expanding our knowledge of how telomere attrition contributes to cellular senescence, organ dysfunction and disease.

Aging and semantic integration in sentence processing: Testing the cognitive workload of wrap-up

The current study investigated the cognitive workload of sentence and clause wrap-up in younger and older readers. A large number of studies have demonstrated the presence of wrap-up effects, peaks in processing time at clause and sentence boundaries that some argue reflect attention to organizational and integrative semantic processes. However, the exact nature of these wrap-up effects is still not entirely clear, with some arguing that wrap-up is not related to processing difficulty, but rather is triggered by a low-level oculomotor response or the implicit monitoring of intonational contour. The notion that wrap-up effects are resource-demanding was directly tested by examining the degree to which sentence and clause wrap-up affects the parafoveal preview benefit. Older and younger adults read passages in which a target word N occurred in a sentence-internal, clause-final, or sentence-final position. A gaze-contingent boundary change paradigm was used in which, on some trials, a non-word preview of word N+1 was replaced by a target word once the eyes crossed an invisible boundary located between words N and N+1. All measures of reading time on word N were longer at clause and sentence boundaries than in the sentence-internal position. In the earliest measures of reading time, sentence and clause wrap-up showed evidence of reducing the magnitude of the preview benefit similarly for younger and older adults. However, this effect was moderated by age in gaze duration, such that older adults showed a complete reduction in the preview benefit in the sentence-final condition. Additionally, sentence and clause wrap-up were negatively associated with the preview benefit. Collectively, the findings from the current study suggest that wrap-up is cognitively demanding and may be less efficient with age, thus, resulting in a reduction of the parafoveal preview during normal reading.

Variable Recruitment in Bundles of Miniature Pneumatic Artificial Muscles and Space Qualified Pneumatic Artificial Muscles

The natural compliance and force generation properties of pneumatic artificial muscles (PAMs) allow them to operate like human muscles in anthropomorphic robotic manipulators. Traditionally, manipulators use a single PAM or multiple PAMs actuated in unison in place of a human muscle. However, these manipulators experience efficiency losses when operated outside their target performance ranges. The unidirectional actuation behavior of a miniature PAM bundle and bidirectional actuation behavior of an antagonistic pair of miniature PAM bundles are characterized and modeled. The results are used to motivate the application of a variable recruitment control strategy to a parallel bundle of miniature PAMs as an attempt to mimic the selective recruitment of motor units in a human muscle to improve the operating efficiency of the actuator. Additionally, the fabrication and quasi-static testing results for PAMs assembled from candidate space qualified bladder and braided sleeve materials for use in space robotics are assessed.

Effects of the spermatozoa: oocyte ratio, water volume and water temperature on artificial fertilization and sperm activation of cascudo-preto

The objective of this study was to evaluate the effects of water volume and water temperature on the sperm motility duration and the number of spermatozoa, and the water volume on the fertilization rates of oocytes of Rhinelepis aspera. Experiments were carried out to evaluate the effect of semen dilutions (1.74×10-5, 1.74×10-4, 1.74×10-3, 1.74×10-2, 1.74×10-1 and 1.00 mL of sperm.mL-1 of water) and water temperature (5, 10, 15, 20, 25, 30, 35, 40, 45, and 50 ºC) on spermatozoa motility duration. In addition, the effects of insemination dose (7×10³, 7×10(4), 7×10(5), 7×10(6) and 7×10(7) spermatozoa.oocyte-1) and water volume (1.0, 30.0, 60.0, 90.0 and 120.0 mL water.2.0 mL-1 oocytes) on the artificial fertilization rates of oocytes were evaluated. The longest sperm motility duration were observed for the semen dilution of 1.74×10-5 mL semen.mL-1 water and in water at 5 ºC. The highest fertilization rates were obtained for insemination doses between 7.00×10³ and 1.23×10(7) spermatozoa. oocyte-1 and water volume of 28.11 mL water.2.0 mL-1 oocytes.

The effect of cross-linker on endogenous enzymatic activity within radicular dentin and bond strength between intraradicular post and radicular dentin

The primary aim was to evaluate the effect of 1-ethyl-3-(3-dimethylamino-propyl) carbodiimide (EDC) on endogenous enzymatic activity within radicular dentin and push-out bond strength of adhesively luted fiber posts, at baseline and after artificial aging. Additionally, the effect of different cementation strategies on endogenous enzymatic activity and fiber post retention was evaluated. The experiment was carried out on extracted human teeth, following endodontic treatment and fiber post cementation. Three cementation strategies were performed: resin cement in combination with etch-and-rinse (EAR) adhesive system, with self-etch (SE) system and self-adhesive (SE) cement. Each of the mentioned strategies had a control and experimental (EDC) group in which root canal was irrigated with 0.3M EDC for 1 minute. The push-out bond strength test was performed 24h after cementation and after 40.000 thermocycles. In order to investigate the effect of EDC and different cementation strategies, in situ zymography analyses of the resin-dentin interfaces were conducted. Statistical analyses were conducted with the software Stata 12.0 (Stata Corp, College Station, Texas, USA) and the significance was set for p<0.05. The results of statistical analysis (ANOVA) showed that the variables “EDC”, “root region” and “artificial aging” significantly influenced fiber posts’ retention to root canal (p<0.05). The highest values were observed in coronal third. The mean values observed after artificial aging were lower when compared to baseline, however EDC was effective in preserving bond strength. The level of enzymatic activity varied between the groups and EDC had a beneficial effect on silencing the enzymatic activity. Within the limitations of the study, it was concluded that the choice of cementation strategy did not influence posts’ retention, while EDC contributed to the preservation of bond strength after artificial aging and reduced enzymatic activity within radicular dentin. In vivo trials are necessary to confirm the results of this in vitro study.

Metodologias para a avaliação do potencial fisiológico de sementes de tabaco

Despite tobacco being a culture propagated by seeds, there is little information concerning tests that allow the distinction of similar germination lots in different levels of vigor. The diversity of cultivars available in the market, and a few peculiarities of the species, such as uneven maturation of the flowers, fruits and seeds, small size and seed dormancy, are considered obstacles for obtaining lots of tobacco of high physiological potential. Thus, this research was developed with the objective of adapting feasibility and vigor tests for evaluating the physiological potential of tobacco seed lots. We used nine lots of tobacco seeds of cultivar CSC 447 and nine lots of seeds of cultivar BAT 2101, belonging to variety groups Virginia and Burley, respectively. Initially, germination test was conducted to characterize the profile of the lots. For determining the feasibility and vigor of the tobacco seeds, germination tests were conducted in distinct temperatures, controlled emergence conditions, electric conductivity, artificial aging and in tetrazolium. For determining the isoenzymatic marker for seed quality, analyses were conducted with enzymes catalase, esterase, malate dehydrogenase and alcohol dehydrogenase. In conclusion, the emergence tests at 25oC and artificial aging at 41oC for 72 hours, are efficient in discriminating the lots of tobacco seeds in different levels of vigor. The electric conductivity and germination tests in different temperatures have distinct responses in relation to the genotype of the tobacco seeds. The tetrazolium test using the methodology with pre-conditioning in 3.5% sodium hypochlorite solution and subsequent emersion in 1.0% tetrazolium solution for 18 hours is efficient for the quick evaluation of the feasibility of tobacco seeds. The analysis of the profiles of enzymes catalase, esterase, malate dehydrogenase and alcohol dehydrogenase is efficient as markers for tobacco seed quality.

Influence of Artificial Accelerated Aging on Dimensional Stability of Acrylic Resins Submitted to Different Storage Protocols

Purpose: The aim of this study was to evaluate the influence of artificial accelerated aging on dimensional stability of two types of acrylic resins (thermally and chemically activated) submitted to different protocols of storage. Materials and Methods: One hundred specimens were made using a Teflon matrix (1.5cmx0.5mm) with four imprint marks, following the lost-wax casting method. The specimens were divided into ten groups, according to the type of acrylic resin, aging procedure, and storage protocol (30 days). GI: acrylic resins thermally activated, aging, storage in artificial saliva for 16 hours, distilled water for 8 hours; GII: thermal, aging, artificial saliva for 16 hours, dry for 8 hours; GIII: thermal, no aging, artificial saliva for 16 hours, distilled water for 8 hours, GIV: thermal, no aging, artificial saliva for 16 hours, dry for 8 hours; GV: acrylic resins chemically activated, aging, artificial saliva for 16 hours, distilled water for 8 hours; GVI: chemical, aging, artificial saliva for 16 hours, dry for 8 hours; GVII: chemical, no aging, artificial saliva for 16 hours, distilled water for 8 hours; GVIII: chemical, no aging, artificial saliva for 16 hours, dry for 8 hours GIX: thermal, dry for 24 hours; and GX: chemical, dry for 24 hours. All specimens were photographed before and after treatment, and the images were evaluated by software (UTHSCSA-Image Tool) that made distance measurements between the marks in the specimens (mm), calculating the dimensional stability. Data were submitted to statistical analysis (two-way ANOVA, Tukey test, p = 0.05). Results: Statistical analysis showed that the specimens submitted to storage in water presented the largest distance between both axes (major and minor), statistically different (p < 0.05) from control groups. Conclusions: All acrylic resins presented dimensional changes, and the artificial accelerated aging and storage period influenced these alterations.