Orientador: Marcelo da Costa Ferreira / Banca: Adeney de Freitas Bueno / Banca: Júlio César Galli / Resumo: Objetivou-se avaliar o espectro, a deposição e cobertura das gotas de pulverização em função de equipamentos, volumes de calda, adjuvantes e dosagem de inseticida na mortalidade de Pseudoplusia includens. No laboratório realizou-se uma aplicação sobre as lagartas com os tratamentos: 2 equipamentos (bico rotativo e hidráulico) 2 volumes de calda (17 e 50 L ha-1 com o bico rotativo e 50 e 100 L ha-1 com o bico hidráulico), 2 dosagens do inseticida endosulfan (0,5 e 1,0 L pc ha-1), e 1 testemunha, seguindo o delineamento inteiramente casualizado. O experimento de campo foi realizado na fazenda da UNESP/Jaboticabal nas safras 2006/07 e 2007/08, em blocos casualizados com: 2 equipamentos (bico hidráulico e rotativo), 3 volumes de aplicação (50; 75; e 100 L ha-1 com o bico hidráulico e 17; 35; e 50 L ha-1 com o bico rotativo), aplicação com e sem adjuvante e 1 testemunha. Avaliou-se o número de lagartas vivas, a cobertura das gotas no papel hidrossensível, a deposição nas folhas de soja, e o espectro das gotas aspergidas utilizando um aparelho medidor de tamanho de partículas em tempo real do Laboratório de Análise do Tamanho de Partículas, do Departamento de Fitossanidade UNESP/Jaboticabal. Verificou-se que o volume de calda pode ser reduzido sem prejudicar o controle de P. includens, a dosagem de 0,5 L pc ha-1 não controla a lagarta-falsa-medideira em laboratório. Em campo, a melhor cobertura, deposição e mortalidade das lagartas foram alcançadas utilizando-se volume de 75 L ha-1 com o bico TT 11001, com a adição de adjuvante. O bico rotativo produz gotas de maior uniformidade e menor porcentagem de gotas suscetíveis à deriva, em relação ao bico hidráulico. A adição de adjuvantes promove gotas de maior tamanho e menos suscetíveis á deriva. / Abstract: The aim of this work was to evaluate the spectrum, the deposition and coverage of droplets of spray due equipment, spray volumes, surfactans and dosage of insecticide in mortality of Pseudoplusia includens. The application in laboratory was done over the caterpillars with the tr~atments: 2 equipment (atomizer and hydraulic nozzles) 2 spray volumes (17 and 50 L ha-1 with the atomizer and 50 and 100 L ha-1 with the hydraulic nozzle), 2 doses of the insecticide endosulfan (0.5 and 1.0 L pc ha-1), and 1 untreated check, following the randomized plots. Was evaluated, the mortality of caterpillars, and the spectrum of droplets. The experiment was carried out in field with randomized blocks: 2 equipment (hydraulic nozzle and atomizer), 3 spray volumes (50, 75, and 100 L ha-1 with hydraulic nozzle and 17, 35 and 50 L ha-1 with atomizer), with and without adjuvant application and 1 untreated check. Was evaluated, the number of alive caterpillars, the coverage of droplets on sensitive paper, deposition on leaves of soybean, and the spectrum of droplets sprayed using in a particle size analyzer in real time. It was observed that spray volume in the laboratory can be reduced without harming the control of P. includens. The dos age of 0.5 L pc ha-1 does not controled satisfactorily the soybean looper caterpillar. On the field, the best coverage, deposition and controlled of caterpillar is achieved using a volume of 75 L ha-1 with hydraulic nozzle TI 11001, with addition of adjuvant. The atomizer produces droplets of more uniformity and less susceptibility to the percentage of droplets to drift, than hydraulic nozzle. The addition of adjuvants results in increase of droplets size and reducing susceptibility to drift. / Mestre

Acoustophoresis is a label free method where the acoustic radiation force is used to manipulate microparticles inside microfluidic channels. The magnitude of the force is dependent of several parameters, which include the density, speed of sound and size of the microparticles, as well as the amplitude of the pressure waves. Recently, acoustophoresis has been used in microfluidics to manipulate microparticles inside moving droplets. In this Master's thesis project, two microfluidic chip designs are used to enrich droplets with polystyrene beads (10 μm in diameter) using acoustophoresis. The microchips have been fabricated with two different fabrication methods; crystalline dependent wet etching and crystalline independent dry etching. In the microchips, water droplets in oil are generated with microparticles suspended in them. By using a channel width that is half a wavelength of the incoming acoustic waves, pressure nodal lines are created in the middle of the channel in which the microparticles align. The droplets then enters a droplet splitting feature, where they are divided into three daughter droplets. Since the majority of the incoming particles are recovered in the center daughter droplet while some of the droplet volume is removed, the center droplet is enriched with the microparticles. For the wet etched design stable droplet splitting was observed when the volumetric flow was 18 μL/min and the incoming droplets had a length-to-width ratio larger than 3. The maximum recovery for this design was 81.1% ± 13.8% with an applied voltage at 10 Vpp. Stable droplet splitting was observed for the dry etched chip at 10.5 μL/min and 18 μL/min at 10 and 20 Vpp, when the incoming droplet had a length-to-width ratio of 3. In this chip the maximum recovery was 93.2% ± 8.3% at the volumetric flow of 10.5 μL/min and an applied voltage of 20 Vpp.

Doctor of Philosophy / Department of Mechanical and Nuclear Engineering / Melanie M. Derby / Power plants are significant water users, accounting for 15% of water withdrawals worldwide. To reduce water usage, compact condensers are required to enable air-cooled condensers and reduce infrastructure costs. Steam flow condensation was studied in 0.952-mm and 1.82-mm hydraulic diameter mini-gaps in an open loop experimental apparatus. The apparatus was validated with single-phase flow. Flow condensation experiments were conducted for a wide range of steam mass fluxes (i.e., 35–100 kg/m²s) and qualities (i.e., 0.2–0.9) in hydrophilic copper and hydrophobic Teflon-coated channels. Water contact angles were 70° and 110° on copper and Teflon, respectively, and in general, filmwise condensation was the primary condensation mode in the hydrophilic channel and dropwise condensation was the primary mode observed in the hydrophobic channel. Pressure drops were reduced by 50–80% in the hydrophobic channels. Condensation heat transfer was enhanced by 200–350% in hydrophobic mini-gaps over hydrophilic mini-gap due to dropwise condensation. Droplet dynamics (e.g., nucleation, coalescence and departure) were quantified during dropwise condensation. A model was created which includes droplet adhesion and drag forces for droplet departure diameters which were then correlated to heat transfer coefficients. An overall mean absolute error of 9.6% was achieved without curve fitting. Noncondensable gases can reduce heat transfer in industrial systems, such as power plants due to the additional layer of thermal resistance from the gas. Condensing steam-nitrogen experiments were conducted for nitrogen mass fractions of 0–30%; the addition of nitrogen reduced heat transfer coefficients by up to 59% and 30% in hydrophilic and hydrophobic mini-gaps, respectively. It was found that during dropwise condensation, the noncondensable layer was perturbed by cyclical droplet motion, and therefore heat transfer coefficients were increased by 2–5 times compared with filmwise condensation of the same mass fraction of nitrogen.

Les anticorps sont des protéines en forme de Y, trouvées comme composant du sérum circulant, qui aident le système immunitaire à cibler et à répondre aux agents pathogènes et aux molécules étrangères, mais peuvent aussi contribuer à la maladie en réagissant aux protéines constitutives. Les anticorps sont produits par des Plasmocytes, qui les sécrètent dans la circulation. Parce qu'il n'y a pas de lien physique entre les plasmocytes et leurs anticorps sécrétés, la détection d'anticorps spécifiques d’un antigène est problématique. Dans cette thèse, j'explore l'utilisation de la microfluidique en gouttelettes pour générer et manipuler des compartiments aqueux homogènes dans lesquels des cellules sécrétant anticorps peuvent être isolées et analysées à haut débit a'échelle d'une seule cellule. Pour caractériser les cellules sécrétant des anticorps à l'intérieur des gouttelettes, j'utilise un nouveau test qui permet d'interroger les cellules en fonction de la spécificité de leur sécrétion. Ces compartiments de gouttelettes peuvent être triés pour le séquençage d'anticorps, ou analysés au cours du temps pour obtenir des informations cinétiques de l'interaction anticorps-antigène à l'intérieur de chaque gouttelette. En utilisant une nouvelle technologie, j'ai pu obtenir le répertoire d'anticorps de souris immunisées contre deux antigènes différents à partir de cellules sécrétant des anticorps spécifiques d’un antigène, avec des capacités égales ou supérieures aux technologies disponibles actuelles. Aussi, j'ai pu suivre le processus de maturation d'affinité des anticorps à l'échelle de la cellule unique, à la fois dans l'immunisation et la maladie auto-immune. Avec ces outils, je démontre comment la microfluidique peut être utilisée pour caractériser les réponses immunitaires et auto-immunes à travers l'évaluation de cellules sécrétant des anticorps. / Antibodies are Y shaped proteins, found as a component of circulating serum, that help the immune system target and respond to pathogens and foreign molecules, but can also contribute to disease when reacting to constitutive self-proteins. Antibodies are produced Plasma Cells, which secrete them into circulation. Because there’s no physical link between Plasma Cells and their secreted antibodies, the detection of antigen-specific antibodies is problematic. In this thesis I explore the use of droplet microfluidics to generate and manipulate homogeneous aqueous compartments in which single antibody secreting cells can be isolated and analyzed in a high-throughput manner. To characterize single antibody secreting cells inside the droplets I use a novel assay that allows to interrogate cells based on the specificity of their secretion. These droplet compartments can be sorted for single cell antibody sequencing, or analyzed over time to obtain kinetic information of the antibody-antigen interaction inside each droplet. Using new established technology I was able to obtain the antibody repertoire of mice immunized against two different antigens from single antigen-specific antibody secreting cells, with equal or better capacities than current available technologies. Also, I was able to follow the affinity maturation process of antibodies at the single-cell level, both in immunization and autoimmune disease. With these tools I demonstrate how microfluidics can be used to characterize the immune and the autoimmune responses through the evaluation of single antibody secreting cells.

The drug development process is notoriously long and expensive. During preclinical studies, inaccurate prediction of pharmacokinetic properties such as the ability of a drug candidate to passively permeate cell plasma membranes contributes to the high failure rate of drug candidates during clinical trials. Passive drug permeability is currently predicted using in vitro techniques such as parallel artificial membrane permeability assays, or PAMPA. In PAMPA, drug transport is predicted between aqueous compartments via a synthetic filter filled with a phospholipid solution in an organic solvent. The lack of translatability of preclinical predictions to humans can be attributed, in part, to lack of biological similarly between models used for permeability prediction and cell plasma membranes in vivo. Here, I demonstrate a new method for pharmacokinetic prediction, built by using droplet interface bilayers (DIBs) as human-mimetic artificial cell membranes. DIBs are bilayer sections created at the interface of two aqueous droplets. In the literature, DIBs have been used as artificial cell plasma membranes to study, for example, electrophysiological properties, protein insertion, water permeability, and molecular transport. DIBs can be formed between droplets of differing composition such that one droplet can be used as a donor compartment and the other as an acceptor compartment for the quantification of molecular transport across the artificial cell membrane. DIBs have previously been used to measure the passive permeability of numerous fluorophores as well as the drugs caffeine and doxorubicin. However, the extent to which DIBs have been tuned to mimic human cell plasma membranes and transport across them is limited. I present here the use of microfluidic platforms for bespoke DIB formation, where variables such as temperature, bilayer composition, and droplet contents are customized to create biomimetic cells-on-a-chip. These artificial cells are then used to measure molecular transport with the aim of predicting permeability. In Chapter 2, I investigate the effectiveness of literature methods for the modification of polydimethylsiloxane (PDMS) microfluidic device channels for aqueous droplet formation and storage. While numerous techniques have been presented as mitigation strategies for common challenges in droplet microfluidics, it is not clear from the literature if any of these methods would be effective or necessary for the formation and analysis of DIBs. With the aim of facilitating aqueous droplet formation, I tested the effect of PDMS silanization using trichloro(1H,1H,2H,2H-perfluorooctyl)silane (PFOS) on surface hydrophobicity and oleophobicity. To assess their effect on reducing the rate of aqueous droplet evaporation, I tested surface treatment of PDMS with Teflon AF or Aquapel. I also tested modifications to the device fabrication process by bonding a glass coverslip to the surface of the device and soaking the device in water overnight. To quantify changes in PDMS surface chemistry, I performed contact angle measurements, aqueous droplet formation experiments, and measurements of droplet size during on-chip storage. I determined that baking PDMS microfluidic devices at 65 C overnight produced channel surfaces which allowed for aqueous droplet formation and storage. In Chapter 3 I present a systematic study on the role of temperature in DIB formation using naturally derived phospholipids. The use of increased temperature to form DIBs using total lipid extracts has previously been demonstrated, but has never before been investigated systematically using naturally derived phospholipids and bespoke formulations thereof. I hypothesized that, in order to form complete phospholipid monolayers and DIBs, the microfluidic device must be held at the phase transition temperature of the phospholipids. Using a custom-built heating platform, I tested DIB formation over a range of temperatures to determine conditions which allowed DIB formation rather than droplet coalescence. I show that temperature is a key parameter for DIB formation using naturally derived phospholipids in a microfluidic device. In Chapter 4, I demonstrate the use of DIBs as a new type of pharmacokinetic compartment model for intestinal absorption. Using three-droplet networks, the components of which were designed to mimic the intestinal space, the enterocyte cytosol, and the blood, I measured fluorescein permeability across intestine-mimetic DIBs. The model was able to predict the transport of fluorescein more accurately than the current state-of-the-art technique, PAMPA. Chapter 5 describes the development of complex DIB models for pharmacologically relevant membranes as well as an investigation into novel methods of drug transport detection on-chip. I created a new DIB model for the small intestine, incorporating more components of the enterocyte plasma membrane such as cholesterol. Measurement of calcein permeability served as a control experiment, as calcein does not cross cell plasma membranes. Measurement of fluorescein permeability yielded a significantly shorter permeation half-life than was determined in Chapter 4, indicating an increase in permeability with the more complex, biomimetic phospholipid formulation. I also developed sex-specific models for intestinal absorption to investigate the effect of sex-based membrane differences on permeability. This relationship has never before been explored in the literature. In comparison to the initial intestinal phospholipid formulation, the sex-specific formulations contained acyl chain tail groups which have been found in different ratios in male and female cells. A significantly longer half-life for fluorescein permeability was found in female intestine-mimetic DIBs, mirroring the slower drug absorption observed in female patients. I also used DIBs to model blood-brain barrier permeability. I demonstrate this application using two different brain lipid extracts, polar and total brain lipids. Polar brain lipids have previously been used in PAMPA to predict blood-brain barrier permeability, but have been found to overpredict the permeation of charged molecules in comparison to custom lipid formulations which mimic the composition of human brain endothelial cells. Permeability measurements in DIBs formed using polar brain lipids gave results which agree with PAMPA, as DIBs formed using polar brain lipids were permeable to fluorescein, but those formed using total brain lipids were not. Blood-brain barrier-mimetic DIBs formed using either lipid extract are impermeable to calcein and FITC-dextrans (both 40 and 500 kDa). I also show the formation of the first DIBs to be created using a total lipid extract from human cells as well as their impermeability to calcein. The extract tested was prepared from testicular Sertoli cells, which exhibit properties similar to the blood-brain barrier, but future work will focus on extracts prepared from human brain endothelial cells. Finally, I explore new options for the on-chip detection of the transport of nonfluorescent molecules. To move away from reliance on fluorescent molecules for permeability measurements, I selected three fluorogenic molecular recognition agents (fluorescamine, Chromeo P540, and DimerDye 4) whose fluorescence signal is activated by amine groups. None of the tested methods proved to be viable in DIBs, potentially due to slow permeation, low quantum yield, and side reactions with phospholipids. Overall, I demonstrate here the microfluidic formation and application of several novel types of biomimetic DIBs to permeability prediction. My work shows that DIBs can be used to predict permeability and mirror effects observed in vivo. Future work will focus on the development of new methods for the detection of drug transport and the application of the pharmacokinetic compartment models presented to predicting drug permeability. Further work using total lipid extracts prepared from human cells will also be vital to enhancing the use of biomimetic DIBs as pharmacokinetic permeability prediction tools. As their biological similarity and capacity to accurately predict transport increase, so will the potential of DIBs to improve the accuracy and translatablity of preclinical drug development. / Graduate / 2023-08-09

No description available.

Pre-ignition is a type of irregular combustion that occurs in boosted direct injection gasoline engines when one or more auto-ignition events occur before to spark ignition. Due to the direct injection of fuel into the cylinder, some liquid fuel may splash off the walls, dragging along lubricating oil. The self-ignition of liquid fuel/lubricant droplets is one of the pre-ignition sources studied. To test this stochastic behavior in a controlled manner, we examined the auto-ignition of a single droplet of a hexadecane-fuel mixture, with hexadecane serving as a surrogate for the lub oil. This experiment involved suspending a single hexadecane-fuel mixture droplet on a thermocouple bead in preheated air at temperatures ranging from 150 to 300 ° C over a wide range of pressures (4-30 bar). Various fuels with RON values ranging from 0 to 120 were blended with hexadecane at varying volume percentages of fuel in hexadecane from 0% to 100% to determine the droplet's time to ignition, denoted by TI. TI was determined by concurrently recording the history of the droplet temperature and imaging it at high speed. The ignition of the droplet is triggered by the self-ignition of the combustible mixture created by the vapor of the hexadecane-fuel mixture reacting with the heated ambient air surrounding the droplet. The increase in RON increased the TI as high RON fuels are difficult to ignite. However, the TI of the mixture depended on the fuel mixture properties even when the RON of the mixture was relatively high. Furthermore, the metal additives were added to the oil surrogate to investigate their effect on getting a pre-ignition event. The lubricant oil additives were phosphate, magnesium, and calcium. These additives were mixed with hexadecane at different concentrations. The experiments were conducted in a constant volume combustion chamber at 300 ⁰C temperature and the pressure was varied from 5 to 15 bar. The resulting TI were then compared with the TI of pure hexadecane. The results showed that addition of phosphate reduces the chances of getting a pre-ignition event, magnesium showed neutral effect while calcium enhanced the chances of getting a pre-ignition event.

An aircraft may experience inlight ice accretion and corresponding reductions in performance and control when the vehicle encounters clouds of super-cooled water droplets. The EADS-IW Surface Engineering Group is investigating passive anti-icing possibilities, such as functional and ice phobic coatings. Ice-resistant coatings require investigating droplet impact on dry surfaces and wet films, including microscopic effects such as droplet splashing. To investigate droplet impacts, a volume of fluid (VOF) flow solver was used for droplets impacting dry and wetted hydrophobic and superhydrophobic surfaces, focusing on meso-scale simulations. The effects of structured, micro-scale surface roughness and the effects of a thin wet film on the surface, corresponding to a saturated surface under high humidity conditions, were investigated. Axisymmetric domains produced acceptable results for smooth, dry surfaces. It was determined that in order to properly predict behavior of droplets impacting surfaces with structured micro-scale roughness, three-dimensional simulations are recommended.

Experiments were performed in which liquid droplets were videographically recorded impacting horizontal superhydrophobic surfaces. The superhydrophobic surfaces were micropatterned with alternating ribs and cavities and coated with a hydrophobic coating. The following surface types were also tested for comparison: smooth uncoated, micropatterned uncoated, and smooth coated surfaces. Three liquid types were used: pure water, ethanol, and a 50/50 water/glycerine mixture. Acquired data demonstrated that the maximum droplet spread diameter exhibited a greater Weber number dependence than that previously reported in the literature. The time delays between impact and maximum spread and between impact and ejection of a vertical jet were characterized, and it was found that experiments with hydrophilic surface behavior follow somewhat different trends than those with hydrophobic behavior, and that there are modest differences between superhydrophobic and hydrophobic surfaces. When analyzing the velocity of the issuing vertical jet, a region of micro-jets was observed with velocities that, under certain conditions, can exceed 15 times the impact velocity. The experimental data acquired were also compared to two recent models from the literature and it was determined that the models do not adequately account for surface anisotropy or apparent slip at the solid-liquid interface. The experiments also showed that instabilities resulting in fingering are dependent upon surface and fluid type, but not contact angle. The onset of peripheral splashing was observed, in general, to occur at a lower Weber number as contact angle increased for the differing surfaces. For surfaces with rib and cavity features, the droplet spread and retraction were generally observed to be asymmetric with spread and retraction faster along the length of the ribs. The occurrence of two-pronged and oscillating jets for water/glycerine tests was also observed for all patterned surfaces. Lastly, an interesting spread pattern with four liquid droplets clustered at about 30° from the perpendicular direction was observed for all fluid types on patterned surfaces for high Weber numbers.

The hydrophobicity of a surface is defined as the degree to which it repels water molecules, and the internal contact angle that the droplet makes with the surface is a measure of the hydrophobicity. Contact angles less than 90° occur on hydrophilic surfaces, while contact angles greater than 90° occur on hydrophobic surfaces. If a surface's contact angle is greater than 120° the surface is commonly defined as superhydrophobic (SH). Superhydrophobicity is accomplished through a combination of microscale surface roughness and water repellant surface chemistry. The roughness creates cavities, or pockets, of vapor underneath the droplet which act to increase the effects of surface tension and lead to increased contact angles. The cavity fraction, F_c, of a surface is a measure of the surface roughness and is defined as the ratio of the projected cavity area to the projected total area of the surface. This thesis investigates the effects of varying cavity fraction, F_c, and substrate temperature, T_s, on heat transfer to evaporating water droplets. Distilled water droplets of nominally 3 mm in diameter were placed on heated SH substrates of varying F_c (0.5, 0.8, and 0.95). A smooth hydrophobic surface was included in the experiments for comparative purposes. The temperature of the surface was held constant at temperatures ranging from 60 to 230°C while the droplet evaporated. Measurements of droplet temperature and size were taken throughout the evaporation process using CCD and infrared camera images. These images were analyzed to yield heat transfer rates for the various surface types and surface temperatures studied. At temperatures below the saturation point of water, average droplet temperatures and heat transfer rates decrease with increasing cavity fraction. Differences in heat transfer rate between substrates increase with substrate temperature. Nusselt number decreases as cavity fraction is increased. Cavity fractions less than about 0.5 show only modest differences in Nusselt number between surfaces. As cavity fraction approaches unity, differences in Nusselt number become amplified between surfaces. At temperatures above the saturation point of water, boiling behavior on SH surfaces deviates dramatically from that of smooth untextured surfaces. Average heat transfer rates decrease with increasing cavity fraction. Nucleate boiling is delayed to highter superheats than normal or is not observed. The Liedenfrost point is advanced to lower superheats as cavity fraction is increased. Similar heat transfer rates are observed beyond the Leidenfrost point.