Thesis (Mech.E)--Massachusetts Institute of Technology, Dept. of Mechanical Engineering, 1980. / MICROFICHE COPY AVAILABLE IN ARCHIVES AND ENGINEERING. / Bibliography: leaves 355-373. / by Martin J. Forbes. / Mech.E

Many daily life products consist of mixtures of oil and water. When an immiscible material is dispersed an interface in-between the two phases is created which gives rise to rheological phenomena which can be exploited for product formulation; this is the case in products such as hand-creams and food products. Furthermore emulsions are used to transport hydrophobic materials, for example, many pharmaceuticals are injected as emulsions into the bloodstream. The performance of such products depends on their microstructure, which is determined by its formulation and how its constituents are mixed together; therefore the microstructure depends on the properties of the dispersed phases, the emulsifier used, the equipment used and its processing conditions. Emulsified products are seldom mono-dispersed due to the complex drop breakup mechanism in the turbulent fields inside the equipment in which the phases are forced together. The chaotic breakup mechanism of highly viscous dispersed phases yield complex and broad drop size distributions (DSD) as a result of the dominating viscous cohesive stresses inside the parent drop. Former studies have used the Sauter mean diameter and/or the size of the largest drop as the characteristic measure of central tendency of the DSD to correlate their results and to prove mechanistic or phenomenological models; however these parameters in isolation are insufficient to characterise the whole DSD of highly polydisperse emulsions. In this dissertation a vast amount of silicon oils of different viscosity were used as dispersed phase to study the effect of various processing conditions and formulations on the resulting DSD. The effect of several formulation and processing parameters were studied for two different mixing devices: stirred vessels and in-line high-shear mixers. (1) For stirred vessels, the effect of stirring speed, continuous phase viscosity and dispersed phase volume fraction were studied in combination with the viscosity of the dispersed phase for steady-state systems. (2) For in-line high-shear mixers a model that links batch and multi-pass continuous emulsification for multimodal DSD was derived from a transient mass balance. Processing parameters such as time and volume, flow rate and number of passes through the mixer, and stirring speed were studied for a wide dispersed phase viscosity range. The analytical methodology implemented included the use of one or more probability density functions to describe the shape of the DSD. The models proposed gave reasonable approximations of the Sauter mean diameter and allowed to study the drop size changes and the relative amount of different types of drops resulting from different breakup mechanisms.

Background: Cerebral small vessel disease (SVD) causes substantial cognitive, psychiatric and physical disabilities. Despite its common nature, SVD pathogenesis and molecular mechanisms remain poorly understood, and prevention and treatment are probably suboptimal. Identifying the genetic determinants of SVD will improve understanding and may help identify novel treatment targets. The aim of this thesis is to better understand genetic associations with SVD through investigating its pathological, radiological and clinical phenotypes. Methods: To unravel the genetic associations with SVD, I used three complementary approaches. First, I performed a systematic review looking at existing intracerebral haemorrhage (ICH) classification systems and their reliability, to help inform future studies of ICH genetics. Second, I performed a series of systematic reviews and meta-analyses, investigating associations between genetic polymorphisms and histopathologically confirmed cerebral amyloid angiopathy (CAA). Third, I performed meta-analyses of existing genome-wide datasets to determine associations of >1000 common single nucleotide polymorphisms (SNP) in the COL4A1/COL4A2 genomic region with clinico-radiological SVD phenotypes: ICH and its subtypes, ischaemic stroke and its subtypes, and white matter hyperintensities. Results: The reliability of existing ICH classification systems appeared excellent in eight studies conducted in specialist centres with experienced raters, although these existing systems have several limitations. In my systematic evaluation of CAA genetics, meta-analyses of 24 studies including 3520 participants showed robust evidence for a dose-dependent association between APOE ɛ4 and histopathological CAA. There was, however, no convincing association between APOE ɛ2 and presence of CAA in a meta-analysis of 11 studies including 1640 participants. Meta-analyses of five studies including 497 participants showed, contrary to an existing popular hypothesis, that while APOE 4 may increase the risk of developing severe CAA vasculopathy, there is no clear evidence to support a role of ɛ2. There were few data about the role of APOE in hereditary CAA, but in the three studies that had looked at this, there was no evidence for an association between APOE ɛ4 and CAA severity. There were too few studies and participants to draw firm conclusions about the effect of non-APOE ε2/ε3/ε4 genetic polymorphisms on CAA, but there were positive associations with TGF-β1, TOMM40 and CR1 genes in four studies. Finally, in my meta-analyses of the COL4A1/COL4A2 genomic region, three intronic SNPs in COL4A2 were associated with SVD phenotypes: significantly with deep ICH, and suggestively with lacunar ischaemic stroke and WMH. Conclusions: I have shown that while existing ICH classification systems appear to have very good reliability, further research is needed to determine their performance in different settings. For large population-based prospective studies of ICH genetics, anatomical systems are likely to be more feasible, scalable and appropriate, although they have limitations and will need to be further developed. Using systematic reviews and meta-analyses, I have confirmed a dose-related association between APOE ɛ4 and histopathological CAA, but also demonstrated that, despite popular acceptance, there is insufficient data to draw firm conclusions about the association with APOE ɛ2. I found some positive associations with CAA in other genes, which merit replication in further larger studies, and showed that there is currently insufficient data about the role of APOE in hereditary CAA. Finally, I identified a novel association between a locus in a known hereditary SVD gene – COL4A2 – and sporadic SVD. This highlights a new and successful approach for selecting candidate genes and can be expanded in future studies to include other known hereditary SVD genes.

Small vessel disease (SVD) is an important cause of stroke, cognitive decline, and age-related disability. The cause of SVD is unknown, increasing evidence from neuropathology and neuroimaging suggests that failure of the blood-brain barrier (BBB) precipitates or worsens cerebral SVD progression and its failure is associated with SVD features such as white matter hyperintensities (WMH), perivascular spaces (PVS) and lacunar infarcts. The BBB change mechanism may also contribute to other common disorders of ageing such as Alzheimer's disease (AD). Magnetic resonance imaging (MRI) has revolutionised our understanding of SVD features. The MRI contributes to better understanding of the SVD pathophysiology and their clinical correlates. The purpose of this project was to better understand the pathogenesis of SVD, which involves improved understanding of BBB structures and pathophysiology and accurate measurement of cerebral SVD imaging characteristics on MRI scans. We aimed to assess (1) structures related to the BBB and factors that affect the BBB; (2) efficient and consistent WMH measurement method; (3) effect of stroke lesions on WMH and cerebral atrophy progression; (4) development and optimisation of computational PVS measurement method; (5) the relationships between PVS and SVD, blood markers, and BBB permeability. Section one describes structures and pathophysiology of the BBB. I reviewed the BBB structural and functional components from the view of neurovascular unit, PVS, and junctional proteins. The PVS part was done in a systematic search. I also reviewed some common stimuli for BBB permeability including inflammation and ischemia. Ischemic triggers for the BBB permeability were summarized systematically. Based on the literatures above, I summarized changes in junctional proteins in ischemia, inflammatory pain and AD models. Section two describes accurate measurement of WMH progression and atrophy. I used data from 100 patients who participated in a stroke study about BBB permeability changes in lacunar versus cortical stroke. To find a most efficient and consistent WMH measurement method, we tested several computational methods and effect of common processing steps including bias field correction and intensity adjustment. To avoid the effect of artefacts, I did a systematic search about artefacts and tested methods of image segmentation to avoid WMH artefacts as much as possible. To investigate the effect of stroke lesions on WMH and atrophy progression, I did the WMH, atrophy segmentation and stroke lesion measurements in a subgroup of 46 patients with follow-up scans, and showed that stroke lesions distorted measurement of WMH and atrophy progression and should be excluded. Section three describes development and optimization of a computational PVS measurement method, which measures the count and volume for PVS based on a threshold method using AnalyzeTM software. We tested the observer variability and validated it by comparison with visual rating scores. We investigated the associations between PVS results with other SVD features (WMH, atrophy), risk factors (hypertension, smoking and diabetes), blood markers, and BBB permeability. In conclusion, MRI is a valuable tool for the investigation of cerebral SVD features and BBB permeability. Exclusions of artefacts and stroke lesions are important in accurate measurement of WMH. PVS are important features of BBB abnormalities, and they correlate and share risk factors with other SVD features, and they should be considered as a marker of SVD and BBB permeability. Further systematic histological and ultrastructural studies of BBB are desirable in understanding the BBB regarding to the different parts of the cerebral vascular tree.

Cerebral small vessel disease (SVD) is a vascular neurodegenerative disease which is the leading cause of vascular dementia and causes 20% of strokes. 20-30% of those over 80 show signs of the disease as white matter hyperintensities on MRI scans, doubling their risk of stroke and trebling their risk of dementia. Sporadic SVD is thought to be caused by hypertension but 30% of sufferers are normotensive and an alternative hypothesis implicates loss of integrity of the blood brain barrier (BBB). To investigate this, I studied brains from normotensive people with early stage SVD and found reduced capillary endothelial claudin-5 (a BBB tight junction protein), more oligodendrocyte precursor cells (OPCs; the precursors to myelinating oligodendrocytes), and more microglia/macrophages compared to controls. Furthermore, in a relevant rat model of spontaneous SVD, the Stroke Prone Spontaneously Hypertensive Rat (SHRSP; disease model; DM) I found that reduced endothelial claudin-5 was the earliest change, appearing at 3 weeks of age, followed by OPC proliferation, appearing at 4 weeks, and then increased number of microglia/macrophages, appearing at 5 weeks. Importantly, all these changes occurred at a young age (< 5 weeks), before any measurable hypertension. These changes were confirmed in an ex vivo slice culture model (i.e. removing blood flow), ruling out direct damage by leakage of blood components through an impaired BBB and suggesting an inherent endothelial cell dysfunction as the primary cause, with secondary BBB defects. This hypothesis of endothelial dysfunction is supported by increased endothelial cell proliferation in both human SVD tissue and the DM rats, and lower levels of endothelial nitric oxide synthase (eNOS) in brains of DM rats. To study this further I isolated primary brain microvascular endothelial cells (BMECs) from DM and control rats and found that those from DM rats formed less mature tight junctions (less membranous claudin-5) than control BMECs. I also found that conditioned media (CM) from DM BMECs causes OPCs in culture to proliferate more and mature less. This indicates that the endothelial dysfunction is inherent to the endothelial cells, rather than induced by other cell types, and through secreted factors causes OPC changes mirroring what is seen in vivo. Using an antibody array, I identified HSP90α as a candidate secreted factor and showed that it is necessary (by blocking the protein in CM) and sufficient (by adding recombinant HSP90α) to induce the maturation phenotype in OPCs, but not the proliferation phenotype. The idea that endothelial dysfunction causes SVD begs the question of what causes endothelial dysfunction, especially in our inbred DM rat strain. To establish this, I reanalysed sequencing data of the DM and control rats from a previously published study, searching for mutations which lead to truncated proteins in genes expressed in brain endothelial cells. We confirmed the candidate gene Atp11b, a phospholipid flippase, was mutated as predicted. I found that knocking down Atp11b using siRNA in a control endothelial cell line caused endothelial dysfunction and a loss of tight junction maturity, and that CM from these cells causes OPCs to proliferate more and mature less, mirroring what we see in primary DM BMECs and suggesting that Atp11b has a key function in promoting normal endothelial function. Furthermore, I showed that knocking down Atp11b causes cells to secrete increased levels of HSP90α. I propose a mechanism whereby ATP11B regulates the retention of HSP90α within endothelial cells, which in turns regulates eNOS levels and activity, as has been shown previously. In summary, this work shows that there are many pre-symptomatic changes which occur in the brain in the development of SVD in DM rats, and that these are ultimately caused by endothelial dysfunction. As these changes are similar to those found in spontaneous human SVD, I propose that endothelial dysfunction is a key mechanism of human SVD, which may in the future lead to new therapies.

Cerebrovascular disease (CVD) can result in cerebral small vessel disease (cSVD) and structural brain changes such as decreased cortical volume, brain atrophy and cerebral infarcts which are major causes of stroke and dementia. CVD is also associated with increased depression and depressive symptoms in later life. Midlife vascular disease and adult socioeconomic status (SES) are well established risk factors but less is known about the effect of factors from earlier in life on CVD and depressive symptoms in later life. A series of systematic reviews of current literature examining early life factors and stroke, cSVD and depression following stroke are presented at the beginning of this thesis. These reviews found that childhood IQ, education and childhood SES were associated with stroke and cSVD in later life. The reviews also found that education level was associated with depression following stroke. However few of the studies adjusted for vascular risk factors and adult SES. Therefore this thesis aimed to investigate associations between birth and childhood factors and cerebrovascular disease and depressive symptoms, after adjustment for vascular risk factors and adult SES, in four community dwelling cohorts: the Stratifying Resilience & Depression Longitudinally (STRADL) cohort (n=280, 45% male, mean age= 62.1 (SD=4.1) years), the Dutch Famine Birth cohort (n= 151, 44% male, mean age 67.6 (SD=0.9) years), the Lothian Birth Cohort 1936 (LBC 1936, n= 865, 50% male, mean age 72.7 (SD=0.7) years), and the Simpson cohort (n=130, 31% male, mean age 78.5, (SD=1.5) years). This Thesis first examined associations between (i) cSVD burden (ii) total and regional brain volumes and (iii) self-reported symptoms of depression and anxiety measured using the Hospital Anxiety and Depression Scale. All analyses were adjusted for age, sex, hypertension, smoking behaviour, adult SES and cognition. Neither cSVD nor brain volumes were associated with symptoms of anxiety. Higher white matter hyperintensity volumes, having one or more cerebral infarct and increased cerebral atrophy were associated with increased depressive symptoms independent of vascular risk factors and adult SES. Secondly, this thesis examined associations between birth and childhood factors and cSVD burden and total and regional brain volumes. Each cohort was analysed individually and then all available data meta-analysed. All analyses were adjusted for age, sex, hypertension, smoking behaviour, adult SES and other early life factors. Meta-analysis found that increasing birth weight was associated with decreased risk of lacunes across all cohorts. Placental weight, which was only available for the Simpson cohort, was associated with decreased risk total cSVD, WMH severity and volume and cerebral infarcts. In the LBC 1936 and Simpson cohort increasing childhood and premorbid IQ and more years of education were associated with fewer cortical infarcts. The association between premorbid and childhood IQ and infarcts was independent of education level. Across three cohorts low education level was associated with more microbleeds. These findings suggest that factors other than traditional vascular risk factors may contribute to cSVD and structural brain changes in later life. Thirdly, this thesis examined associations between birth and childhood factors and self-reported symptoms of depression and anxiety measured using the Hospital Anxiety and Depression Scale (HADS) and the Quick Inventory of Depressive Symptoms (QIDS-16). All analyses were adjusted for age, sex, hypertension, smoking behaviour, adult SES and cognition In the Dutch Famine Birth Cohort people born before the famine had lower scores of depression and anxiety on the HADS compared to those exposed to famine in early gestation and those conceived after the famine. In the LBC 1936 increasing ponderal index was associated with lower depressive symptoms, increasing childhood and premorbid IQ were associated with lower symptoms of anxiety and depression. Lower educational attainment and some indicators of childhood SES were associated with higher symptoms of depression and anxiety. Overall results suggest that early life factors, particularly childhood IQ, may contribute to structural brain changes and symptoms of depression and anxiety in later life, independent of vascular risk factors and other early life factors. Efforts to understand factors which may contribute to late life health, from the earliest stages of life, are important and may be used to inform changes in social policy. The effect sizes and potential impact of these findings suggest that larger sample sizes with more vascular disease and more depression are needed to robustly test these associations.

Abnormalities in the vascular pattern of a retina, such as morphologic changes in vessel shape, branching pattern, width, tortuosity, or the appearance of retinal lesions, may be associated with the occurrence of retinopathies or cardiovascular diseases. Thus, an automated quantitative analysis of changes in vessel morphology may help indicating the clinical signs of aforementioned retinopathies, describing their early occurrence or severity. The responses obtained from different types of retinal vessels, i.e., arteries and veins, may be variable to retinopathies and their measurement may lead to a more precise diagnosis compared to that by the average response accounted for the entire vessel network. I propose a set of automated methods in order to analyze the retinal vessel network and to quantify its morphologic properties with respect to arteries and veins, in two-dimensional color fundus images. The analytical methods include; 1) Forma- tion of a well connected vessel network, 2) Structural mapping of a vessel network, 3) Artery-venous classification, and 4) Blood vessel hemorrhage detection. The quan- tification methods include vessel morphology analysis based on the measurement of tortuosity, width, branching angle, branching coefficient, and fractal dimension. The aforementioned morphologic parameters are measured with respect to arteries and veins separately in a vessel network. The methods are validated with the manually annotated retinal fundus images as a ground truth. The major contribution of this thesis includes the development of automated methods for; 1) Identification and separation of retinal vessel trees for individual vessel analysis, 2) Automated quantification of morphologic characteristics of retinal vessels for quick and precise measurement, 3) Automated quantification of vessel morphology with respect to arteries and veins, and 4) Analysis of two datasets, a) malarial retinopathy subject dataset, b) longitudinal study dataset. The ability of the automated methods to quantify the retinal vessel specific properties may enable the individual vessel analysis as an alternative to a time- consuming and subjective clinical evaluation, or to a quantitative morphology char- acterization averaged over the entire vessel network. The objective evaluation may indicate the progression of retinopathies precisely and may help characterizing nor- mal and abnormal vascular patterns with respect to arteries and veins. This may enable a quick diagnosis, treatment availability, prognosis, and facilitation of clinical health-care procedures in remote areas.

Ischemic stroke is the leading cause of disability world-wide and affects over 800,000 people per year in the United States. The majority of these strokes are ischemic due to a blockage of blood flow to the brain. Damage to the brain occurs at the onset of stroke, neuronal cell death is irreversible and therefore, quick treatment to remove blockage is critical factor in the recovery from stroke. Mechanical thrombectomy as a treatment for ischemic stroke provides an ideal opportunity to collect blood distal and proximal to the cerebral thrombus to examine neurochemical changes occurring during stroke. The purpose of this dissertation was to explore the trajectory of neurochemical changes that occur in response to ischemic stroke during the first 72 hours and the physiological response from stroke patients to improve stroke outcomes. The specific aims were to: 1) to determine whether venous blood gases predict infarct volume and/or mortality in acute ischemic stroke in young male rats; 2) determine whether venous blood gases predict infarct and edema volume, and/or mortality in acute ischemic stroke in aged male and female rats; 3) compare the presence and relative concentrations of acid/base and electrolytes in static blood distal to thrombus and in peripheral blood drawn from adults who received thrombectomy for ischemic stroke and identify associations to postreperfusion functional outcomes. Specific Aim One was addressed by evaluation of young (three-month old) Sprague-Dawley rats that underwent permanent or transient middle cerebral artery occlusion (MCAO). Pre- and post-MCAO venous samples from permanent and transient models provided pH, carbon dioxide, oxygen, bicarbonate, glucose, hematocrit, hematocrit, and electrolyte values of ionized calcium, potassium and sodium. The analyses indicated that mean differences in the blood gas and electrolytes between pre- to post-MCAO and pH and iCa2+ were predictors of infarct volume in the permanent MCAO model. The second aim was addressed by evaluation of aged (18 month old) male and female rats pre-MCAO, post-MCAO, and at 72 hours of permanent MCAO venous blood gas samples (pH, carbon dioxide, oxygen, bicarbonate, glucose, hematocrit, hematocrit, and electrolyte concentrations of ionized calcium, potassium and sodium). Changes in pH (from pre-MCAO to post-MACO and post-MCAO to 72 hours) and changes in Na+ and iCa2+ (from post-MCAO to 72 hours) were predictors of infarct volume and edema volume, respectively in both sexes. Cox regression revealed there was a 3.25 times increased risk for mortality based on changes (cut-off range within -2.00 to - 7.00) in bicarbonate levels (pre- to post-MCAO). The third aim was addressed by evaluation of acid/base balance (pH, carbon dioxide, oxygen, bicarbonate, ionized calcium, potassium and sodium) of ischemic stroke patients who underwent mechanical thrombectomy. Our results suggests sex differences matter in ischemic stroke populations. Significant differences occur within proximal blood between the sexes. Additionally, females had approximately 2.5 hour increased time between stroke symptom onset to thrombectomy completion time (described as infarct time). Changes in bicarbonate and base deficit were predictors of infarct time, but only in our female population.

While trade relations between French colonists and indigenous peoples in New Orleans are well documented, there have been few in depth studies utilizing archaeological sites in the city to illuminate the ways in which such relations shaped the day to day lives of the peoples involved. This work has attempted to elucidate trade practices between these groups by utilizing archaeological data uncovered at 810 Royal Street during excavations from 2015 through 2018. A collection of hand-built ceramics typically associated with indigenous peoples found in French colonial contexts on the site may help explicate the nature of trade occurring within the city and the ways in which this trade was reflective of larger patterns of urban colonial adaptation and creolization. This work seeks to illuminate the motivations behind such trade and the ways in which economic motives and individual self-interests drove colonists to undermine the original French designs for the city.

Chronic obstructive pulmonary disease (COPD) is a type of obstructive lung disease characterized by chronically poor airflow, which is the result of breakdown of lung tissue (known as emphysema) and small airways disease. It typically worsens over time. Most treatments are limited to the management of symptoms, which makes early detection more valuable to treat the disease etiology itself. With the advancement of computed tomography (CT), it is able to provide high resolution structural and functional imaging to distinguish the lung anatomic structures, as well as characterize their changes over time. Previously, the majority of CT-based measures have focused on quantifying the extent of airway and parenchymal damage. Recent studies suggests that pulmonary vascular dysfunction is an early lesion in COPD and associated with an emphysematous phenotype. Few studies attempted to quantify pulmonary vessel morphology and compared those measures across COPD groups. However, the scope of examined vascular structures in these studies was limited, majorly due to the lack of a standardized method to quantify a broad range of vascular structures. In this thesis, we propose to use anatomically defined airway branches as references to locate and morphologically quantify central pulmonary arteries in different lung regions. Although pulmonary vessel trees have complex topologic and geometric structures, airway tree possesses much simpler and consistent branching patterns and standardized anatomic nomenclatures are available up to sub-segmental levels. It is also well-known that airway and arterial branches have a unique pairing that is established by their spatial proximity and parallel configuration. Therefore, anatomically labeled airway tree provides a robust benchmark to locate consistent arterial segments for both intra- and inter-subjects. New methods have been developed for quantitative assessment of arterial morphology matched and standardized by associated airways at different anatomic branches. First, the skeletons of airway and vessel trees are generated to provide simple and hierarchical representations. Then, topologic and geometric properties of airways and arteries, such as distance, orientation and anatomic positon information, are explored to locate the target arterial segments. Finally, the morphologic properties, e.g. cross-sectional area, of target arterial segments are robustly computed. The developed methods in this thesis provides a standardized framework to assess and compare the vascular measurements in intra- and inter- subjects from a broad range of vessel branches in different lung regions. The work also serves as a practical tool for large longitudinal or cross-sectional studies to explore the pulmonary vessel roles played at the early stage of COPD. The major contribution of this thesis include: (1) developing two novel skeletonization methods that are applicable to airway and pulmonary vessel trees; (2) developing a semi-automatic method to locate and quantify central pulmonary arterial morphology associate to anatomic airway branches; (3) developing a fully automatic method to identify and reconstruct central pulmonary arterial segments associated to anatomic airway branches and quantify their morphology; (4) validating the methods using computerized phantoms, physical phantoms and human subjects; (5) applying the developed methods to two human lung disease studies.