Dry powder inhalers (DPIs) are a common therapeutic modality for lung diseases such as asthma, but they are also used to treat systemic diseases such as diabetes. Advantages of DPIs include their portable design and low manufacturing costs. Another advantage of DPIs is their breath activation, which makes them popular among patients. In a passive DPI drug is only released when the patient inhales. When the patient inhales, air flows through the device. The flow of air entrains a dry powder formulation inside the device and carries it to the lung. Currently, no DPI exists which can deliver drug independent of the patient to the desired target site in the lung. This is because drug release depends on the patient’s inhalation manoeuvre. To maximize the effect of the treatment it is necessary to optimize DPIs to achieve drug delivery that (A) is independent of the inhalation manoeuvre and (B) is targeted to the correct site in the lung. Therefore, this thesis aims to apply numerical and experimental methods to optimize DPIs systematically. First, two clinically justifiable cost functions have been developed corresponding to the DPI design objectives (A) and (B). An Eulerian-Eulerian (EE) computational fluid dynamics (CFD) approach has then been used to optimize a DPI entrainment geometry. Three different optimized entrainment geometries have been found corresponding to three different therapeutic applications. Second, the CFD approach has been validated experimentally. This is the first experimental study to validate an EE CFD approach for DPI modelling. Third, a personalized medicine approach to DPI design has been proposed. The development of this approach makes it possible to achieve the design objectives for patients with highly different lung functions. Finally, an adaptive DPI with a variable bypass element has been developed. This DPI achieves design objectives (A) and (B) for patients with highly different lung functions with a single device. In contrast to the personalized medicine approach, there is no need to select the optimal amount of bypass, since the device adapts automatically.

This study is an exposition of research into the effect of operational efficiency to global competitiveness, with the Richards Bay Dry Bulk Terminal (DBT) being the case study. The Richards Bay DBT is the main dry bulk terminal in the South African ports, operated by Transnet Ports Terminals (TPT), which is a division of the state-owned Transnet Ltd. Because of the strategic position of the DBT as the main dry bulk terminal of the country and also that it is a state owned entity, its competitiveness or otherwise is a reflection of South Africa the country as a trade destination. The research seeks to investigate the problem of the declining volumes handled by the DBT, which is traced to the possible operational inefficiencies and is likely to impede competitiveness of the DBT. While this study will not propose the ultimate solutions to the DBT’s problems, it will identify the root causes and symptoms of the problems, which will ultimately guide what needs to be tested as a better solution. Based on this, it is argued that this topic should generate a lot of interest to the stakeholders both locally and internationally.

Typescript (photocopy). / Digitized by Kansas Correctional Industries

Cryoconite holes are vertical columns of meltwater within the shallow subsurface of glaciers. In the McMurdo Dry Valleys (MDV) of Antarctica cryoconite holes are a source of meltwater and harbor microbial communities in an otherwise arid environment with low biologic activity. The holes form as sediments on the ice surface, which are darker than the surrounding ice, are preferentially heated by solar radiation. The warm sediments melt the underlying ice and migrate downwards. An ice lid forms, isolating them from the below-freezing atmosphere enabling them to remain thawed. In this study, field observations, laboratory experiments, and numerical modeling are used to characterize the fundamental variables controlling cryoconite hole development. Field and laboratory results show that solar radiation drives cryoconite hole melting by controlling the energy available to the cryoconite and to warm the surrounding ice. Holes deepen further in warmer ice. Laboratory results show that at temperatures of -10º C at least 405 (W m-2) are needed to warm the cryoconite sufficiently to melt surrounding ice. Numerical modeling shows that increased radiation flux into the subsurface and warmer air temperatures cause cryoconite to descend deeper and the meltwater-filled holes to enlarge, while increased surface ablation decreases their average depth. Cryoconite holes thaw sooner and refreeze later when the optical properties of the ice facilitate greater radiation transmission. Cryoconite warms the ice significantly more than ice without cryoconite. Within the melt-filled hole, the heat capacity of the water keeps the surrounding ice warm for several weeks after the cryoconite-free ice has cooled. The cryoconite itself is last to completely freeze.

Extreme environments provide a unique source of often highly adapted and tolerant organisms. Research on organisms in these habitats has led to the discovery of novel and useful compounds and may assist in understanding the impact of global change on biodiversity. The Dry Valleys of Eastern Antarctica are vast, ice-free regions believed to be the coldest, driest desert on Earth. Despite these harsh conditions, there is an increasing amount of evidence demonstrating that the soil ecosystems of the Dry Valleys sustain a wide diversity of microorganisms. The research presented is an inter-valley comparison study which aims to scrutinize microbial communities and environmental factors driving their distribution in the Dry Valleys. Automated ribosomal intergenic spacer analysis (ARISA) was used to provide a snapshot of bacterial and cyanobacterial communities living in the mineral sands in Miers Valley, Beacon Valley, Upper Wright Valley and at Battleship Promontory. Rigorous analysis of physico-chemical differences between the soils of these four valleys was undertaken in hope to understand the environmental parameters driving the distribution and biodiversity of microbial communities present. Multivariate statistical analysis and ordination of ARISA and physico-chemical data revealed that bacterial communities from each valley form distinctive clusters. Conversely, cyanobacterial communities showed less diversity and a more even distribution between valleys.

Isotropic ice samples containing measured concentrations of solutes and debris similar to basal material found in several cold-based glaciers in the McMurdo Dry Valleys, Antarctica, were manufactured in a laboratory and tested for peak shear strength at constant strain rates with a direct-shear device. The shear tests show that differences in rheology and shear strength appear to be related to impurity content and concentration. Debris-laden ice becomes more ductile with greater concentrations of solutes, whereas, low solute-concentrations and high debris-concentrations are associated with increases in shear strength and brittle behaviour. Stress exponents from Glen�s flow law calculated for isotropic solute and debris-laden ice ranged between 4 and 5, leading to the conclusion that higher rates of deformation may be expected in dirty basal ice than predicted for glacial ice models that use stress exponents where, n = 3. Observations of both natural and synthetic samples tested over a range of temperatures between -25�C and -5�C showed that natural basal ice samples containing high solute and debris concentrations were highly sensitive to temperature change. These tests showed an approximate 10 % loss in shear strength for every 1�C increase in temperature between -25�C and -10�C. In addition, contrasts in rheology and rates of deformation within basal ice are responsible for the development of debris-laden ice structures in the basal zones of cold-based glaciers that flow over unconsolidated substrates. As layered sedimentary bedding was preserved in frozen blocks within the deforming basal ice of several of these glaciers, the evidence suggests that at some point each glacier has interacted with its bed and entrained portions of the substrate material. Empirical shear strength data and observations of rheological changes attributed to composition together with evidence acquired during fieldwork in Antarctica help to support the argument that cold-based glaciers flowing over unconsolidated sediment are capable of affecting geomorphic change. Hence, isotropic ice models that exclude basal processes may need to be adjusted, especially where small increases in the temperature of the basal zones of cold glaciers may occur. In conclusion, palaeo-climate inferences based purely upon small amounts of geomorphic evidence, which suggest warmer climate conditions, may need to be re-evaluated in order to portray more accurate renditions of formerly glaciated landscapes.

The composition of ice from four case studies based on the facies, solute, stable isotope, and debris content reveals compositional differences reflective of different modes of ice formation. In Southern McMurdo Sound, there is a distinctive geochemical signature that differentiates between meteoric-origin and marine-origin ice. Analysis of the basal ice of three glaciers from the McMurdo Dry Valleys shows that liquid water does contribute to its formation. The basal ice sequences are structurally and compositionally different and are reflective of different modes of formation or entrainment active at the glacier margins. In the cases of the Rhone and Wright Lower glaciers marginal sediments and liquid water are key to understanding the accretion of debris-rich ice and both have basal facies consistent with refreezing in subzero conditions. The liquid water is formed by ephemeral melt during the summer. In the Rhone Glacier, melt water refreezes on the apron and is entrained into the advancing glacier. By contrast, by the Wright Lower Glacier adjacent streams or ponds saturate unconsolidated sediments which are entrained during ice advance. In the Taylor Glacier, the basal ice is comprised of a thick sequence of intercalated layers of clean clear ice and fine-grained debris layers. These laminated facies have a solute composition consistent with evaporites formed from a relict seawater intrusion. The combination of entrained debris, high solutes and laminations is consistent with interaction at the glacier bed and regelation. Interpreting empirically derived co-isotopic slopes is problematic, as highlighted in the case study of the Taylor Glacier where laminated facies have all the hallmarks of refrozen ice, yet plot on a co-isotopic slope that is typically interpreted as meteoric. Similarly, ice from the McMurdo Ice Shelf shows a clear difference in absolute isotope values which is interpreted as being refrozen from seawater, yet its co-isotopic plot is statistically indistinguishable from the meteoric water line. The ice compositional approach has highlighted several shortcomings. Firstly, solutes deposited in inland areas have limited solute pathways and do not distinguish between different types of ice but are useful in distinguishing between marine and continental salts. Secondly, co-isotopic analysis to reconstruct freezing history is dependent on statistically-derived interpretations which do not explain slopes that lie between physically-based models of meteoric and freezing slopes. In empirical studies, slopes between 5 and 8 are common, and are probably cosmopolitan samples. Finally, ice composition is inconsistent between similar ice types in the McMurdo Dry Valleys, as similar facies have different ice compositions, and origins. This underlines the problem with the premise that structurally similar ice facies are formed by the same process.

Introduction Menopause which is defined as a permanent physiological, or natural, cessation of menstrual cycle, plays an important role in the development of ocular surface dryness symptoms and there is an increased prevalence of dry eye in women, especially those aged over 50. Despite the high prevalence of dry eye in post-menopausal women (PMW), very few studies have been undertaken to understand dry eye disease in a group of PMW who are not on Hormone Replacement Therapy (HRT). Studies in the past on PMW have primarily focused on the relationship between HRT and dry eye. Hence, a series of studies were undertaken to understand the clinical aspects of dry eye and their relationship to a variety of tear film components, in a group of PMW with and without symptoms of dry eye. The specific aims of each chapter were as follows: • Chapter 4: To characterize symptoms of dry eye using questionnaires, namely Ocular Surface Disease Index Questionnaire© (OSDI) and the Indiana Dry Eye Questionnaire (DEQ). • Chapter 5: To characterize clinical signs and symptoms in participants who present with and without symptoms of dry eye. • Chapter 6: To compare tear osmolality and ferning patterns in participants with and without dry eye symptoms. • Chapter 7: To investigate the potential relationship between subjective symptoms and clinical signs with tear film lipocalin and lysozyme concentrations in participants with and without dry eye symptoms. • Chapter 8: To optimize a technique for the isolation of total RNA (ribo nucleic acid) and total protein derived from conjunctival epithelial cells collected via conjunctival impression cytology (CIC). • Chapter 9: To quantify the expression of MUC1 (mucin1) and MUC16 (mucin16) mRNA and protein and to investigate the potential relationship between mucin expression and tear film breakup time in a group of participants with and without dry eye symptoms. Methods • Chapter 4: Participants were categorized as being symptomatic or asymptomatic of dry eye based on their response to the OSDI questionnaire. These results were then compared to the DEQ, which has questions related to the frequency of ocular surface symptoms and their diurnal intensity. • Chapter 5: Non invasive tear breakup time (NITBUT) was evaluated using the ALCON Eyemap®. Tear volume was assessed using the Phenol Red Thread (PRT) test and bulbar conjunctival hyperemia was measured using objective (SpectraScan PR650© Spectrophotometer) and subjective (slit lamp) methods. • Chapter 6: Tears were collected via capillary tube. A freezing point depression osmometer was used to measure the osmolality of the tear film. The tear ferning test was performed and evaluated for the quality of ferning, based on the Rolando grading system. • Chapter 7: Tears were collected via capillary tube and an eye wash method. Tear lysozyme and lipocalin concentrations were determined via Western blotting. • Chapter 8: CIC was collected using either Millipore (MP) or Poly Ether Sulfone (PES) membranes. RNA and protein isolation was performed using two different RNA isolation techniques. Two methods of protein isolation from CIC discs were evaluated. RT-PCR of mRNA for MUC1 and western blotting of lipoxygenase type 2 protein (LOX2) was performed to confirm the collection of intact RNA and total protein respectively. • Chapter 9: Tears were collected via capillary tube and an eye wash method. CIC was collected using MP membrane. Expression of MUC1 and MUC16 mRNA was assessed via real time PCR. Expression of both membrane-bound and soluble MUC1 and MUC16 were quantified via Western blotting. Results • Chapter 4: The OSDI total score and sub scores for the Non Dry Eye (NDE) and Dry Eye (DE) groups were significantly different (NDE =7.43 ± 7.71 vs DE = 24.87 ± 13.89; p<0.001). The DEQ scores showed that the DE group exhibited a higher frequency and intensity of symptoms than the NDE group, which worsened as the day progressed (p<0.001). • Chapter 5: The DE group exhibited a significantly shorter NITBUT (5.3 ± 1.7 vs 7.0 ± 2.7 secs; p=0.0012). Tear volume was significantly lower for the DE group (19.3 ± 5.1mm vs. 16.3 ± 5.6mm; p=0.031). Bulbar hyperemia was significantly higher in the DE group for both objective (u’ = 0.285 ± 0.006 vs. 0.282 ± 0.006; p=0.005) and subjective techniques (48.4 ± 10.0 vs 40.6 ± 10.4; p=0.0011). • Chapter 6: Osmolality values in DE individuals were significantly higher than the NDE (328.1 ± 20.8 vs. 315.1 ± 11.3 mOsm/kg; p = 0.02). There was a significant difference between the DE and NDE participants for the ferning patterns (p = 0.019). No significant correlation between tear osmolality and tear ferning was noted (DE: r = 0.12; p > 0.05, NDE: r = -0.17; p > 0.05). • Chapter 7: No difference in tear lysozyme or lipocalin concentration was found between DE and NDE groups, irrespective of tear collection method. Method of collection significantly influenced absolute concentrations (p<0.008). • Chapter 8: There was no significant difference between the two procedures used to isolate RNA and protein from CIC membranes (p>0.05). Total RNA yield was greater with the MP membrane. The mean yield of protein extracted from MP membrane using the two protein isolation techniques also did not show a significant difference. • Chapter 9: No difference was found in the expression of either MUC1 or MUC16 protein or mRNA expression between symptomatic DE and NDE (p>0.05). Weak correlations were found between the NITBUT values compared with either soluble or membrane bound MUC1 and MUC16 expression. Conclusions • Chapter 4: Questionnaires are useful tools to symptomatically divide participants into dry eyed and non dry eyed candidates. However, the questionnaire used to categorise patients can impact on the outcome variables determined. • Chapter 5: Post-menopausal women with dry eye symptoms demonstrate shorter NITBUT, lower tear volume and increased bulbar conjunctival hyperemia than those who have no symptoms. • Chapter 6: Tear osmolality in DE is higher than in NDE. There is a tendency towards less ferning in persons over 50 years of age, regardless of their symptoms. • Chapter 7: Comparison of clinical data with lipocalin and lysozyme concentrations failed to reveal statistically significant correlations. The concentration of either protein was not associated with tear stability or secretion. • Chapter 8: The total RNA yield was greater with the MP membrane. RNeasy Mini (RN) (Qiagen) method is recommended due to enhanced speed as well as on-column isolation and DNase digestion capabilities. CIC with MP membranes followed by immediate freezing and then extraction and processing facilitates the collection of total protein from human conjunctival cells. • Chapter 9: No difference was found in the expression of either MUC1 or MUC16 protein or mRNA expression between symptomatic PMW and asymptomatic controls.

Sjogren’s syndrome (SS) is a systemic autoimmune disease that presents to eye care practitioners with the hallmark symptom of “dry eye.” Stratifying dry eye patients as Sjogren’s positive or negative is a critical differential diagnosis, as SS patients have numerous systemic complications and a forty times greater risk of developing lymphoma. As such, management of this relatively common dry eye sub-population requires specialized care. Since a firm diagnosis requires testing that is both invasive and expensive, patients should be protected from these tests if they are not warranted. In this thesis, studies were therefore undertaken to determine if SS dry eye could be differentiated from other forms of dry eye using two methods: 1) standard clinical tests used in a multi-disciplinary Sjogren’s syndrome clinic and 2) subsequent biological evaluation of collected tear samples and cells from the ocular surface. The former would allow eye care practitioners to conduct appropriate tests and pose suitable questions to ifferentiate these subgroups, and the latter might serve in the future as a relatively non-invasive quantitative means of differentiating such groups through biomarkers.

Introduction: Tear film osmolality is a product of the varying concentrations of dissolved solutes (proteins, lipids and mucins) in the tear fluid. Research suggests that a hyperosmotic tear film is a trait common to all forms of dry eye, and it may be the driving force causing the discomfort, ocular surface damage and inflammation found in both evaporative and tear deficient forms of dry eye disease. Tear film osmolality has been proposed to be the “gold standard” diagnostic test for the evaluation of dry eye disease, as a distinct separation between tear film osmolalities in normal and dry-eyed (aqueous deficient or evaporative) populations has become evident. Historically, tear film osmolality could only be measured in a laboratory setting and required a highly skilled technician to use the instrumentation. The recent development of easy-to-use, small volume osmometers has made it possible for tear film osmolality to be measured clinically. As these instruments are quite new, there has been very little research completed with them. Therefore, a series of studies was conducted to investigate the utility of one of these new osmometers – the Advanced Instruments Model 3100 Nanolitre Osmometer. The specific aims of each chapter were: - Chapter 3: To determine if the Advanced Instruments Model 3100 Nanolitre Osmometer was capable of quantitatively measuring tear film osmolality in a normal population, using 0.5μL tear samples. - Chapter 4: Previous studies have shown the Advanced Instruments Model 3100 Nanolitre Osmometer not significantly different from another commercially available osmometer (Wescor Vapor Pressure Osmometer) for the measurement of human tears. This chapter examined the repeatability of the new instrument over multiple measurements on the same sample and over multiple days. - Chapter 5: To determine if tear film osmolality values varied significantly over the course of a normal working day in a population that was primarily free from symptoms of dry eye. - Chapter 6: To investigate the relationships between tear film osmolality and other commonly used clinical tests for dry eye disease. The clinical tests examined included various questionnaires designed to assess patient symptoms (Single Item Dry Eye Questionnaire (SIDEQ), the Ocular Surface Disease Index (OSDI), and the McMonnies Dry Eye Questionnaire (MMDEQ) and a linear analogue comfort scale (LACS)), a non-invasive tear break-up time test (NIBUT), and examination of ocular surface redness and tear ferning (TF). Secondarily to determine if the other clinical tests demonstrated significant diurnal variations over the course of a normal working day. - Chapter 7: To measure tear film osmolality in a population with mild to moderate symptoms of dry eye disease, and to compare this value with the osmolality of a population of age-matched controls without the disease. Secondarily, to investigate the relationship between tear film osmolality and patient comfort in a population with mild to moderate symptoms of dry eye disease. Methods: - Chapter 3: Tears were collected from 40 volunteer participants with a capillary tube. Some participants were non-contact lens wearers (Non-CL), while others wore either soft or rigid contact lenses (CL). Tear film osmolality was measured with the Advanced Instruments Model 3100 Nanolitre Osmometer. - Chapter 4: Tears were collected from 10 volunteer participants using two different collection techniques. Collections were repeated on three separate days (6 study visits total); three osmolality measurements per collection were taken using the Advanced Instruments Model 3100 Nanolitre osmometer. - Chapter 5: Tears were collected from 40 volunteer participants in two separate studies (n=80 in total). Tears were collected with a capillary tube three times a day (morning, mid-day and afternoon), on two separate days (6 study visits total). Tear film osmolality was measured with the Advanced Instruments Model 3100 Nanolitre Osmometer. - Chapter 6: Clinical tests were administered and tear samples were collected using a capillary tube from 40 volunteer participants. Measurements were taken three times a day (morning, mid-day and afternoon), on two separate days (6 study visits total). Tear film osmolality was measured with the Advanced Instruments Model 3100 Nanolitre Osmometer. - Chapter 7: Participants were classified as either having dry eye disease (DE) or not having dry eye disease (NDE) based on a clinical examination that included a case history, phenol red thread test and biomicroscopy (white light and sodium fluorescein assessment). Tear samples were then collected from all participants using a capillary tube and tear film osmolality was measured with the Advanced Instruments Model 3100 Nanolitre Osmometer. Participants also completed the SIDEQ, the OSDI, and the MMDEQ. Results: - Chapter 3: The mean tear film osmolality of the population was 298.7±11.4mOsm/Kg. CL wear (soft or rigid) did not appear to have a significant effect on tear film osmolality (CL: 298.5±11.2mOsm/Kg vs. Non-CL: 298.9±11.5mOsm/Kg), although this study was not designed to specifically look at the effects of contact lens wear on tear film osmolality. - Chapter 4: There was reasonably good concordance between measurements of tear film osmolality taken with the Advanced Instruments Model 3100 Nanolitre Osmometer (intraclass correlations range from 0.6497 (F= 0.0582) to 0.9550 (F = 0.5893)). Repeatability appeared to be affected by significant changes in ambient humidity (>10% per day). Concordance was similar with both sampling techniques. - Chapter 5: In the first study, no significant diurnal change in tear film osmolality was found (p>0.05), although a significant difference in measurements taken on Day 1 compared to Day 2 was found (p=0.040). When the first and last 10 participants enrolled were compared, the difference between days was present in the first 10 participants, but not in the last 10; it is likely that the investigator underwent a learning process during the period of the study, and that reflex tearing occurred more often in the early portion of the study compared with the latter portion. In the second study, no significant diurnal change in tear film osmolality was found (p>0.05) and no significant difference in measurements taken on Day 1 compared to Day 2 was found (p>0.05). When tear film osmolality was compared with the number of hours participants were awake, no significant correlation was found (r = 0.07044). - Chapter 6: Significant correlations were not found between tear film osmolality and SIDEQ (r = 0.1347), OSDI (r = 0.0331), MMDEQ (r = 0.2727), LACS (r = -0.1622), NIBUT (r = -0.2280), subjectively graded redness (r=-0.2280), or objectively measured redness (r = 0.1233). A weakly significant correlation was found between TF and tear film osmolality (r = 0.3978). None of the clinical measures (LACS, NIBUT, subjective or objective redness or TF) varied significantly over the course of the day. - Chapter 7: Tear film osmolality was higher in both the right (DE = 311.1±12.4mOsm/Kg, NDE = 306.2±11.2mOsm/Kg) and left eyes (DE = 313.2±11.9mOsm/Kg, NDE = 304.0±7.5mOsm/Kg) of participants, but the difference was only statistically significant in the left eye. Tear film osmolality did not correlate significantly with DE patient symptoms using any of the questionnaires (SIDEQ, OSDI, MMDEQ). Conclusions: - Chapter 3: The Advanced Instruments Model 3100 Nanolitre Osmometer appeared to be capable of measuring tear film osmolality in a normal population. Our population mean was slightly lower than what is reported to be normal (305mOsm/Kg), but it still fell within the range of values reported as normal (297 – 318mOsm/Kg). - Chapter 4: The Advanced Instruments Model 3100 Nanolitre Osmometer demonstrated reasonably good repeatability for the measurement of human tear samples. Unfortunately, the instrumentation appeared to be affected by dramatic weather changes. Maintaining the instrument in a humidity controlled environment may resolve this problem. - Chapter 5: Tear film osmolality did not appear to vary significantly over a normal working day. Inducing reflex tearing, perhaps with an unskilled investigator collecting the tears, can be a significant source of error (as demonstrated in the first study). - Chapter 6: Tear film osmolality did not correlate well with other clinical instruments designed to assess either patient symptoms or signs of dry eye disease in a normal population. Tear film osmolality and tear ferning did demonstrate a weakly significant positive correlation. None of the clinical measures assessed demonstrated a significant diurnal variation over the course of a normal working day. - Chapter 7: Tear film osmolality appeared to be higher in participants with mild to moderate symptoms of dry eye when compared with age matched, asymptomatic controls. Tear film osmolality did not correlate well with patient symptoms in a population of mild to moderate severe dry eyed individuals.