Introduction: Aging has been shown to diminish bone blood flow in rats and humans. The purpose of this study was to determine whether blood flow to regions of the femur perfused primarily through the principal nutrient artery (PNA) are diminished with aging and whether this putative reduction in flow is associated with impaired endothelium‐dependent vasodilation. Materials and Methods: Blood flow was measured in conscious young adult (4–6 mo old) and aged (24–26 mo old) male Fischer‐344 rats using radiolabeled microspheres. Endothelium‐dependent vasodilation of the PNA was assessed in vitro using acetylcholine (ACh), whereas the contribution of the NO synthase (NOS) and cyclooxygenase (COX) signaling pathways to endothelium‐dependent vasodilation was determined using the NOS and COX inhibitors L‐NAME and indomethacin, respectively. Results: Femoral blood flow in the aged rats was 21% and 28% lower in the proximal and distal metaphyses, respectively, and 45% lower in the diaphyseal marrow. Endothelium‐dependent vasodilation was reduced with old age (young: 83 ± 6% maximal relaxation; aged: 62 ± 5% maximal relaxation), whereas endothelium‐independent vasodilation (sodium nitroprusside) was unaffected by age. The reduction in endothelium‐dependent vasodilation was mediated through impairment of the NOS signaling pathway, which resulted in lower NO bioavailability (young: 168 ± 56 nM; aged: 50 ± 7 nM). Conclusions: These data show that reductions in metaphyseal bone and diaphyseal marrow perfusion with old age are associated with diminished endothelium‐dependent vasodilation through an impairment of the NOS mechanism. Such age‐related changes in bone perfusion and vascular NO signaling could impact clinical bone loss, increase risk of fracture, and impair fracture healing in the elderly.

Vascular disease is the leading cause of mortality and morbidity in the western world, and account for the 1 of every 3 death’s in the US, but a cure for vascular disease is yet to be realized. Hematopoietic stem progenitor cells (HSPCs) are mobilized from bone marrow and have the innate propensity to accelerate vascular repair by reendothelialization and revascularization of ischemic areas. The vasoreparative ability of HSPCs is largely due to their capacity to home to the areas of hypoxia and their sensitivity to hypoxia plays a critical role in the vasoreparative functions of these cells. The discovery of vasoreparative potential of HSPCs resulted in a breakthrough approach of cell-based therapies for the treatment of ischemic vascular diseases. However, success of this approach is essentially dependent on the number of cells that could be collected from an individual. Therefore, novel mechanism-based strategies are needed to enhance the outcomes of autologous cell-based therapies in poor mobilizers and older adults. Recent evidence of a potential role of the vasoprotective axis of the renin angiotensin system (RAS) in HSPCs functions offers a breakthrough. Angiotensin-(1-7), the primary mediator of the protective functions which acts on Mas receptor (MasR), is generated by angiotensin converting enzyme-2 (ACE2). In this study, we tested the effects of hypoxia on stimulation of vasoreparative potential of HSPCs and in upregulation of ACE2 and MasR. Importantly, we delineated the molecular mechanism of hypoxic exposure in regulation of ACE2 and MasR in a HIF1α- dependent manner and hypoxic exposure induced shedding of the membrane bound ACE2 in HSPCs. We used luciferase, a reporter assay, cell-based assays, gene/protein expression studies and pharmacological strategies in human and mouse HSPCs to test our hypotheses. To verify the biological significance of hypoxia, we performed in vivo studies in mice and humans, which recapitulated the in vitro observations on vascular protective axis of RAS in HSPCs. Collectively, these studies provided mechanistic insights into hypoxic regulation of vascular protective axis of RAS in HSPCs and also provided compelling evidence for the clinical use of hypoxia as a promising approach for enhancing the vasoreparative outcomes of cell-based therapies. / American Heart Association grant, 13SDG16960025 / National Institutes of Health, National institute of Aging (NIA), 1R01AG056881

Cette thèse porte sur l’étude de la séquence d’imagerie IRM IVIM (« Intravoxel incoherent motion »). Cette séquence permet l’étude des microvaisseaux sanguins tels que les capillaires, artérioles et veinules. Pour être sensible seulement aux groupes de spins non statiques dans les tissus, des gradients de diffusion sont ajoutés avant et après l’impulsion 180° d’une séquence d’écho de spin. La composante du signal correspondant aux spins qui diffusent dans le tissu peut être séparée de celle des spins en mouvement dans les vaisseaux sanguins qui est appelée signal IVIM. Ces deux composantes sont pondérées par f IVIM qui représente la fraction volumique du sang à l’intérieur du tissu. Le signal IVIM est en général modélisé par une fonction mono-exponentielle (ME) caractérisée par un coefficient de pseudo-diffusion D*. Nous proposons un modèle IVIM bi-exponentiel formé d’une composante lente caractérisée F slow et D* slow qui correspondrait aux capillaires comme dans le modèle ME, et d’une composante rapide caractérisée par F fast et D* fast qui correspondrait à des vaisseaux plus gros comme des artérioles et veinules. Ce modèle a été validé expérimentalement et des informations supplémentaires ont été obtenues en comparant les signaux expérimentaux avec un dictionnaire de signaux IVIM simulés numériquement. L’influence de la séquence d’impulsions, du temps de répétition et du temps d’encodage de diffusion a également été étudiée. Enfin, la séquence IVIM a été appliquée à l’étude d’un modèle animal de la maladie d’Alzheimer. / This PhD thesis is centered on the study of the IVIM (“Intravoxel Incoherent Motion”) MRI sequence. This sequence allows for the study of the blood microvasculature such as the capillaries, arterioles and venules. To be sensitive only to moving groups of spins, diffusion gradients are added before and after the 180° pulse of a spin echo (SE) sequence. The signal component corresponding to spins diffusing in the tissue can be separated from the one related to spins travelling in the blood vessels which is called the IVIM signal. These two components are weighted by f IVIM which represents the volume fraction of blood inside the tissue. The IVIM signal is usually modelled by a mono-exponential (ME) function and characterized by a pseudo-diffusion coefficient, D*. We propose instead a bi-exponential IVIM model consisting of a slow pool, characterized by F slow and D* slow corresponding to the capillaries as in the ME model, and a fast pool, characterized by F fast and D* fast, related to larger vessels such as medium-size arterioles and venules. This model was validated experimentally and more information was retrieved by comparing the experimental signals to a dictionary of simulated IVIM signals. The influence of the pulse sequence, the repetition time and the diffusion encoding time was also studied. Finally, the IVIM sequence was applied to the study of an animal model of Alzheimer’s disease.

This thesis sought to evaluate whether the increased risk of heat-related illness observed in black-African descendants stems from impairements in local- and whole-body heat loss responses in this ethnic group. To evaluate this, in separate studies local- (study 1) and whole–body (study 2) heat loss responses were compared in young men (18-30 y) of black-African (n=21) and Caucasian (n=21) descent, matched for physical characteristics and fitness and born and raised in the same temperate environment. In study 1, we compared nitric oxide-dependent skin blood flow and sweating responses in young men of black-African (n=10) and Caucasian (n=10) descent during rest, exercise, and recovery in the heat. Both groups rested for 10-min, and then performed 50-min of moderate-intensity exercise at 200 W/m2, followed by 30-min of recovery in hot-dry heat (35°C, 20% RH). Local cutaneous vascular conductance (CVC%max) and sweat rate (SR) were measured at two forearm skin sites treated with a) lactated-Ringer (Control), or b) 10 mM NG-nitro-L-arginine methyl ester (L‐NAME, NO synthase-inhibitor). L-NAME significantly reduced CVC%max throughout rest, exercise, and recovery in both groups (both p<0.001). However, there were no significant main effects for the NO contribution to CVC%max between groups (all p>0.500). L-NAME significantly reduced local SR in both groups (both p<0.050). The NO contribution to SR was similar between groups such that L-NAME reduced SR relative to control at 40 and 50 min into exercise (both p<0.050). In study 2, we assessed whole-body total heat loss (evaporative + dry heat exchange) in black-African (n=11) and Caucasian (n=11) men using direct calorimetry. Participants performed three, 30-min bouts of semi-recumbent cycling at fixed metabolic heat productions (and therefore matched heat loss requirements between groups) of 200 (light), 250 (moderate), and 300 W/m2 (vigorous), each followed by 15-min recovery, in dry heat (40°C, ~13% relative humidity). Across all exercise bouts, dry (p=0.435) and evaporative (p=0.600) heat exchange did not differ significantly between groups. As such, total heat loss during light, moderate and vigorous exercise was similar between groups (p=0.777), averaging ((mean (SD)); 177 (10), 217 (13) and 244 (20) W/m2 in men of black-African descent, and 172 (13), 212 (17) and 244 (17) W/m2 in Caucasian men. Accordingly, body heat storage across all exercise bouts (summation of metabolic heat production and total heat loss) was also similar between the black-African (568 (142) kJ) and Caucasian groups (623 (124) kJ; p=0.356). This thesis demonstrates that ethnicity does not influence NO-dependent cutaneous vasodilation and sweating in healthy, young black-African descent and Caucasian men during exercise in the heat. Furthermore, we extend upon these observations by showing no differences in whole-body dry and evaporative heat exchange and therefore body heat storage.

PURPOSE: This study aims to quantify the retinal vascular blood flow in eyes affected by unilateral central retinal vein occlusions (CRVO) or branch retinal vein occlusions (BRVO). We created and explored a new, unitless metric for the severity of these diseases: relative blood flow (RBF). We then contextualized RBF in terms of patient demographics, ocular presentation and other systemic conditions, as well as explored its efficacy as a predictor of future outcomes. METHODS: Data was collected from 20 control subjects and 32 patients with clinically diagnosed retinal vein occlusions (15 CRVO and 17 BRVO). Laser speckle flowgraphy was then used to quantify retinal vascular blood flow in terms of mean blur rate, a metric shown to be highly heterogeneous between patients but fairly consistent in intra-patient repeated measurements over time. After confirming this and establishing a strong correlation between a healthy patient’s two eyes, we used an RVO patient’s fellow eye as a nondiseased expectation and presented relative blood flow as the ratio between their diseased and healthy eye. We then correlated this data with demographic variables and disease characteristics from patients’ medical history. RESULTS: We found an average blood flow decrease of 26% in CRVO eyes relative to healthy eyes in the same patients and an average decrease of 7% in BRVO eyes. In CRVO, duration of occlusion, central macular thickness, intraocular pressure, diabetes, previous laser and injection treatments, and an injection within three months after blood flow measurement were significantly associated with relative blood flow. In BRVO, no demographic variables or disease characteristics were significantly associated with relative blood flow. CONCLUSIONS: Relative blood flow represents a promising new, consistent and informative metric for quantifying the severity of unilateral retinal vein occlusions. With both descriptive and predictive properties in eyes with CRVO, future work should explore its great potential.

With high energy consumption and a low capacity for energy storage, the brain is highly dependent on a continuous supply of oxygen and nutrients from the bloodstream. Ischemic stroke, caused by the occlusion of a cerebral blood vessel, compromises cerebral blood flow (CBF), resulting in detrimental effects on brain homeostasis, vascular function, and neuronal health. Sex differences in ischemic stroke are known, with women having lower rates of stroke due to a protective role of estrogens on vascular health, and more severe strokes following reduced estrogen production after menopause. Rho-associated protein kinase (ROCK), an important regulator of vascular tone, also regulates vascular function in a sex-specific manner, and its deletion is neuroprotective following ischemic stroke. The current study explores the overlapping roles of ROCK and endogenous hormone influence on the acute CBF response to a photothrombotic (PT) model of ischemic stroke in mice. CBF was measured following stroke in the somatosensory cortex in mice with a heterozygous deletion of the ROCK2 isoform (ROCK2+/-) and in wild-type (WT) littermates. To remove endogenous hormones, male mice were gonadectomized (Gdx) and female mice were ovariectomized (Ovx), and control animals received a sham surgery (“intact”) prior to stroke induction. Intact WT males showed a delayed CBF drop compared to intact WT females, where peak drop in CBF wasn’t observed until 48 hours following stroke. Gonadectomy in males did not alter this response, however ovariectomy in females produced a “male-like” response. ROCK2+/- males also showed such phenotypic response, and Gdx did not alter this response, suggesting ROCK2 deletion or endogenous male hormones do not alter CBF response in males in this stroke model. Alternatively, intact ROCK2+/- females showed a striking difference in CBF values compared to intact WT females, where they displayed higher CBF values immediately post-stroke and also showed a peak drop in CBF at 48 hours post-stroke. Ovx did not change the CBF response in ROCK2+/- females. Overall, there is a marked difference between males and females in their acute CBF responses to PT stroke, which appears to be mediated by endogenous female sex hormones and ROCK2. All groups except for intact WT females show a delayed drop in CBF values, reaching a maximal drop in CBF at 48 hours following stroke induction. This may be due to hyperreactivity of female platelets and upregulation of RhoA/ROCK signaling in female platelets. Further research is required to confirm this speculation. This study reveals important sex-differences and the involvement of ROCK2 in acute CBF responses to PT stroke in mice.

No description available.

Indiana University-Purdue University Indianapolis (IUPUI) / Over 5 million older adults have Alzheimer's disease (AD) in the US, and this number is projected to double by 2050. Clinical trials of potential pharmacological treatments for AD have largely shown that once cognitive decline has occurred, targeting AD pathology in the brain does not improve cognition. Therefore, it is likely that the most effective treatments for AD will need to be administered before cognitive symptoms occur, necessitating a biomarker for the early, preclinical stages of AD. Cerebral blood flow (CBF) is a promising early biomarker for AD. CBF is decreased in individuals with AD compared to their normally aging counterparts, and it has been shown that CBF is altered in mild cognitive impairment (MCI) and earlier stages and may occur prior to amyloid or tau aggregation. In addition, CBF can be measured using arterial spin labeled (ASL) MRI, a noninvasive imaging technique that can be safely repeated over time to track prognosis or treatment efficacy. The complex temporal and spatial patterns of altered CBF over the course of AD, as well as the relationships between CBF and AD-specific and -nonspecific factors, will be critical to elucidate in order for CBF to be an effective early biomarker of AD. Here, we begin to characterize the relationships between CBF and risk factors, pathologies, and symptoms of AD. Chapter 1 is a systematic review of published literature that compares CBF in individuals with AD and MCI to CBF in cognitively normal (CN) controls and assesses the relationship between CBF and cognitive function. Chapter 2 reports our original research assessing the relationships between CBF, hypertension, and race/ethnicity in older adults without dementia from the the Indiana Alzheimer’s Disease Research Center (IADRC) and Alzheimer’s Disease Neuroimaging Initiative (ADNI). Chapter 3 reports our original research assessing the relationships between CBF and amyloid beta and tau aggregation measured with PET, as well as whether hypertension or APOEε4 positivity affects these relationships, in older adults without dementia from the IADRC. Chapter 4 reports our original research assessing the relationship between the spatial distribution of tau and subjective memory concerns. / 2023-05-24

Orientador: Franciele Marques Vanderlei / Resumo: Estudos sobre o treinamento resistido de baixa intensidade associado à restrição de fluxo sanguíneo (RFS) atualmente tem ganhado destaque, porém ainda existem lacunas que podem ser exploradas em relação aos seus efeitos fisiológicos quando associado a treinamentos excêntricos, principalmente na força e hipertrofia muscular. Objetivo: Realizar uma revisão sistemática com metanálise e um estudo aleatorizado controlado a fim de esclarecer os reais efeitos do TRBI associado à RFS nos desfechos de força, hipertrofia muscular e desempenho funcional, e se a RFS quando associada a treinamentos excêntricos de baixa e alta intensidade é capaz de aumentar as respostas desses desfechos dos extensores de joelho em homens adultos saudáveis. Métodos: A revisão sistemática foi registrada no PROSPERO (número do registro #CRD42018104065). Os estudos foram selecionados por meio de sete bases de dados. Todos os estudos incluídos foram avaliados quanto à qualidade metodológica, utilizando a Escala PEDro, bem como foi avaliada a qualidade da evidência. Somente ensaios clínicos randomizados que avaliaram a eficácia do treinamento com RFS na força e hipertrofia muscular dos músculos extensores de joelho em adultos do sexo masculino, saudáveis e/ou fisicamente ativos, com idade de 18 a 35 anos, contra algum grupo de intervenção foram considerados elegíveis. Todas as metanálises foram conduzidas por meio do software Review Manager – RevMan e descritos como diferenças médias padronizadas (standardized ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Studies on low intensity resistance training associated with blood flow restriction (RFS) are currently gaining prominence, however there are still gaps that can be explored in relation to their physiological effects when associated with eccentric training, especially in muscle strength and hypertrophy. Objective: To carry out a systematic review with meta-analysis and a randomized controlled study in order to clarify the real effects of TRBI associated with RFS on the outcomes of strength, muscle hypertrophy and functional performance, and whether RFS when associated with low and high intensity eccentric training is able to increase the responses of these knee extensor outcomes in healthy adult men. Methods: The systematic review was registered in PROSPERO (registration number # CRD42018104065). The studies were selected through seven databases. All included studies were assessed for methodological quality, using the PEDro Scale, as well as the quality of evidence was assessed. Only randomized controlled trials that assessed the effectiveness of RFS training on muscle strength and hypertrophy of knee extensor muscles in healthy, male and / or physically active adults, aged 18 to 35, against any intervention group were considered eligible. All meta-analyzes were conducted using the Review Manager - RevMan software and described as standardized mean differences (SMD) or mean differences (mean difference - MD) with 95% confidence intervals (CI). The randomized controlled study ... (Complete abstract click electronic access below) / Mestre

Indiana University-Purdue University Indianapolis (IUPUI) / Glaucoma is a multifactorial ocular disease progressively leading to irreversible blindness. There is clear evidence of correlations between alterations in ocular hemodynamics and glaucoma; however, the mechanisms giving rise to these correlations are still elusive. The objective of this thesis is to develop mathematical models and methods to help elucidate these mechanisms. First, we develop a mathematical model that describes the deformation of ocular structures and ocular blood flow using a reduced-order fluid-structure interaction model. This model is used to investigate the relevance of mechanical and vascular factors in glaucoma. As a first step in expanding this model to higher dimensions, we propose a novel energy-based technique for coupling partial and ordinary differential equations in blood flow, using operator splitting. Next, we combine clinical data and model predictions to propose possible explanations for the increase in venous oxygen saturation in advanced glaucoma patients. We develop a computer-aided manipulation process of color Doppler images to extract novel waveform parameters to distinguish between healthy and glaucomatous individuals. The results obtained in this work suggest that: 1) the increase in resistance of the retinal microcirculation contributes to the influence of intraocular pressure on retinal hemodynamics; 2) the influence of cerebrospinal fluid pressure on retinal hemodynamics is mediated by associated changes in blood pressure; 3) the increase in venous oxygen saturation levels observed among advanced glaucoma patients depends on the value of the patients’ intraocular pressure; 4) the normalized distance between the ascending and descending limb of the ophthalmic artery velocity profile is significantly higher in glaucoma patients than in healthy individuals.