A comprehensive review of the nasal route of administration, in particular the nasal drug delivery system has been presented. The physicochemical properties, mode of action and pharmacology of H1-receptor antagonists, in particular cyclizine HCl, have been highlighted. The techniques for the assessment of toxicity (in-vitro ciliary beat frequency (CBF) studies for human nasal explants and morphology studies of the rat nasal mucosa), synthesis of cyclizine lactate, solubility studies of both cyclizine HCI and cyclizine lactate, viscosity determination of the gel formulated and assessment of the deposition and distribution of the hydroxypropylmethyl cellulose (HPMC) dispersions within the human nasal cavity model were conducted. In this study, preliminary studies on the toxicity of the various formulation components (excipients and active ingredient) were carried out. Results from these studies indicated that for both the excipients and the drug, pH significantly affects the ciliary motility hence all ciliary beat frequency determinations were conducted at nasal pH. Furthermore, effects of the various concentrations (0.0625%(w/v), 0.125%(w/v), 0.25%(w/v), 0.5%(w/v) and l%(w/v)) of the excipients on ciliary motility were investigated. Transmission electron microscopy (TEM) studies proved useful in evaluating the integrity and changes in the surface morphology of the rat nasal mucosa post treatment with the various excipients (carboxymethyl cellulose, hydroxypropylmethyl cellulose, trimethyl chitosan 36.3% DQ, Carbopol P934 and polysorbate-80) at varying concentrations. Of the excipients investigated, hydroxypropylmethyl cellulose (HPMC) showed ciliofriendliness since there was no apparent ultra structural damage, although a slight decrease in ciliary beat frequency (CBF) was observed at the highest viscosity. Moreover, hydroxypropylmethyl cellulose (HPMC) is said to be a bioadhesive excipient, which would therefore confer its bioadhesive properties to the intranasal preparation to enhance the retention time between the absorbing mucosa and the drug and hence increase nasal drug absorption. This excipient was therefore selected as the ideal for use in the formulation of the intranasal preparation. The aqueous solubility of a drug plays an important role in nasal administration since it is required that the drug component be applied in a limited volume of about 200pl. To enhance the aqueous solubility of the sparingly water-soluble cyclizine HCl, a lactate salt was synthesised and characterised. This compound was found to be highly soluble in water. The intranasal preparation was therefore manufactured using the lactate form of cyclizine. A single blind study was conducted to determine and compare the pharmacokinetic parameters for both Valoid oral tablets containing 100mg cyclizine HCl (reference drug) and cyclizine lactate intranasal preparation 125mglml (study drug). The results obtained indicated a significant improvement in the bioavailability of cyclizine. For oral administration Cmax = 200.79ng/ml at tmax = 5.57h and for the intranasal preparation Cmax = 5354.22ng/ml at tmax = 1.59h. A 19.2-fold increase in drug bioavailability was observed after intranasal administration (AUCin = 122860.70ng/ml/h) compared with oral administration (AUCpo = 5943.48ng/ml/h). This enhanced bioavailability through nasal administration indicated that enhanced nasal drug absorption and hence increased bioavailability not only depends on the favourable anatomical and physiological characteristics of the nasal mucosa but possibly on the inherent physico-chemical characteristics of the drug molecule and the formulation components. Thus chemical modification of the sparingly water-soluble cyclizine HCl to the highly water-soluble cyclizine lactate facilitated the dissolution of more solute in a limited volume of solvent. This new feature therefore may have impacted positively to the transport of cyclizine across the nasal mucosa. Furthermore, the hydroxypropylmethyl cellulose (HPMC), component of the formulation, could have conferred its mucoadhesive properties to the preparation. Perhaps it increased the retention time of the dosage form within the nasal passages through bond formation with the nasal mucosa thereby increasing the contact time between the absorbing mucosa and the dosage form. This interaction between the mucoadhesive and the nasal mucosa may have resulted in the modification of tissue permeability (possibly transient opening of the tight junctions) and eventual increase in the drug penetration/absorption. / Thesis (Ph.D. (Pharmaceutics))--North-West University, Potchefstroom Campus, 2004.

A comprehensive review of the nasal route of administration, in particular the nasal drug delivery system has been presented. The physicochemical properties, mode of action and pharmacology of H1-receptor antagonists, in particular cyclizine HCl, have been highlighted. The techniques for the assessment of toxicity (in-vitro ciliary beat frequency (CBF) studies for human nasal explants and morphology studies of the rat nasal mucosa), synthesis of cyclizine lactate, solubility studies of both cyclizine HCI and cyclizine lactate, viscosity determination of the gel formulated and assessment of the deposition and distribution of the hydroxypropylmethyl cellulose (HPMC) dispersions within the human nasal cavity model were conducted. In this study, preliminary studies on the toxicity of the various formulation components (excipients and active ingredient) were carried out. Results from these studies indicated that for both the excipients and the drug, pH significantly affects the ciliary motility hence all ciliary beat frequency determinations were conducted at nasal pH. Furthermore, effects of the various concentrations (0.0625%(w/v), 0.125%(w/v), 0.25%(w/v), 0.5%(w/v) and l%(w/v)) of the excipients on ciliary motility were investigated. Transmission electron microscopy (TEM) studies proved useful in evaluating the integrity and changes in the surface morphology of the rat nasal mucosa post treatment with the various excipients (carboxymethyl cellulose, hydroxypropylmethyl cellulose, trimethyl chitosan 36.3% DQ, Carbopol P934 and polysorbate-80) at varying concentrations. Of the excipients investigated, hydroxypropylmethyl cellulose (HPMC) showed ciliofriendliness since there was no apparent ultra structural damage, although a slight decrease in ciliary beat frequency (CBF) was observed at the highest viscosity. Moreover, hydroxypropylmethyl cellulose (HPMC) is said to be a bioadhesive excipient, which would therefore confer its bioadhesive properties to the intranasal preparation to enhance the retention time between the absorbing mucosa and the drug and hence increase nasal drug absorption. This excipient was therefore selected as the ideal for use in the formulation of the intranasal preparation. The aqueous solubility of a drug plays an important role in nasal administration since it is required that the drug component be applied in a limited volume of about 200pl. To enhance the aqueous solubility of the sparingly water-soluble cyclizine HCl, a lactate salt was synthesised and characterised. This compound was found to be highly soluble in water. The intranasal preparation was therefore manufactured using the lactate form of cyclizine. A single blind study was conducted to determine and compare the pharmacokinetic parameters for both Valoid oral tablets containing 100mg cyclizine HCl (reference drug) and cyclizine lactate intranasal preparation 125mglml (study drug). The results obtained indicated a significant improvement in the bioavailability of cyclizine. For oral administration Cmax = 200.79ng/ml at tmax = 5.57h and for the intranasal preparation Cmax = 5354.22ng/ml at tmax = 1.59h. A 19.2-fold increase in drug bioavailability was observed after intranasal administration (AUCin = 122860.70ng/ml/h) compared with oral administration (AUCpo = 5943.48ng/ml/h). This enhanced bioavailability through nasal administration indicated that enhanced nasal drug absorption and hence increased bioavailability not only depends on the favourable anatomical and physiological characteristics of the nasal mucosa but possibly on the inherent physico-chemical characteristics of the drug molecule and the formulation components. Thus chemical modification of the sparingly water-soluble cyclizine HCl to the highly water-soluble cyclizine lactate facilitated the dissolution of more solute in a limited volume of solvent. This new feature therefore may have impacted positively to the transport of cyclizine across the nasal mucosa. Furthermore, the hydroxypropylmethyl cellulose (HPMC), component of the formulation, could have conferred its mucoadhesive properties to the preparation. Perhaps it increased the retention time of the dosage form within the nasal passages through bond formation with the nasal mucosa thereby increasing the contact time between the absorbing mucosa and the dosage form. This interaction between the mucoadhesive and the nasal mucosa may have resulted in the modification of tissue permeability (possibly transient opening of the tight junctions) and eventual increase in the drug penetration/absorption. / Thesis (Ph.D. (Pharmaceutics))--North-West University, Potchefstroom Campus, 2004.

[pt] Este trabalho apresenta um método teórico e experimental para medir Perda Dependente de Polarização (PDL) em sistemas ópticos. Este método basea-se na forma como duas componentes de polarização ortogonais da luz moduladas em amplitude por duas frequências distintas são acopladas no detector após a transmissão pelo dispositivo sob caracterização. No presente trabalho é realizado um estudo acerca das propriedades da luz e das técnicas tradicionais utlizadas para medir PDL. Antes da caracterização das medidas de PDL, foi realizado o alinhamento do dispositivo de teste (DUT), ou seja, do sistema que vai emular a PDL, a fim de garantir que o mesmo estivesse com as perdas ópticas minimizadas. Em seguida, conectou-se o DUT à configuração montada no laboratório, sendo realizado os ajustes de polarização e supressão das portadoras ópticas. Neste método é possível calcular o valor da PDL e da na orientação na Esfera de Poincaré. / [en] This work presents a theoretic and experimental method for Polarization Dependent Loss (PDL) measurement in optical systems. This method is based on how two orthogonal polarization components both modulated in amplitude by two different frequency are coupled in the detector. In this present work a study is performed about light properties and traditional techniques used to peform PDL measurement. Before the characterization of PDL measures the alignment of the device under test (DUT) was performed to make sure that optical losses were minimized in the system which will emulate the PDL. Then, the DUT was connected to the experimental setup to optical carriers suppression and polarization alignment. In this method is possible to calculate the PDL value and its orientation on the Poincaré sphere.

Eukaryotic organisms rely on the coordinated beating of motile cilia for a multitude of fundamental reasons. In smaller organisms, such as Paramecium and the single cell alga Chlamydomonas reinhardtii, it is a matter of propulsion, to swim towards a higher concentration of nutrients or away from damaging environments. Larger organisms use instead the coordinated motion of cilia to push fluid along an epithelium: examples common to mammals are the circulation of cerebrospinal fluid in the brain, the transport of ovules in the fallopian tubes, and breaking the left/right symmetry in the embryo. Another notable example, and one that is central to this thesis, is mucociliary clearance in human airways: A carpet of motile cilia helps keeping the cell surface free from pathogens and foreign particles by constantly evacuating from lungs, bronchi, and trachea a barrier of mucus. The question of how motile cilia interact with one another to beat in a coordinated fashion is an open and pressing one, with immediate implications for the medical community. In order for the fluid propulsion to be effective, the motion of cilia needs to be phase-locked across significant distances, in the form of travelling waves (``metachronal waves''). It is still not known how this long-range coordination emerges from local rules, as there is no central node regulating the coordination among cilia. In the first part of this thesis I will focus on studying the coordination in carpets of cilia with a top-down approach, by proposing, implementing, and applying a new method of analysing microscope videos of ciliated epithelia. Chapter 1 provides the reader with an introduction on motile cilia and flagella, treating their structure and motion and reporting the different open questions currently tackled by the scientific community, with particular interest in the coordination mechanisms of cilia and the mucociliary clearance apparatus. Chapter 2 introduces Differential Dynamic Microscopy (DDM), a powerful and versatile image analysis tool that bridges the gap between spectroscopy and microscopy by allowing to perform scattering experiments on a microscope. The most interesting aspects of DDM for this work are that it can be applied to microscope videos where it is not possible to resolve individual objects in the field of view, and it requires no user input. These two characteristics make DDM a perfect candidate for analysing several hundred microscope videos of weakly scattering filaments such as cilia. In Chapter 3 I will present how it is possible to employ DDM to extract a wealth of often-overlooked information from videos of ciliated epithelia: DDM can successfully probe the ciliary beat frequency (CBF) in a sample, measure the direction of beating of the cilia, and detect metachronal waves and read their direction and wavelength. In vitro ciliated epithelia however often do not show perfect coordination or alignment among cilia. For the analysis of these samples, where the metachronal coordination might not be evident, we developed a new approach, called multiscale DDM (multiDDM), to measure a coordination length scale, a characteristic length of the system over which the coordination between cilia is lost. The new technique of multiDDM is employed in Chapter 4 to study how the coordination among cilia changes as a response to changes in the rheology of the mucous layer. In particular, we show that cilia beating under a thick, gel-like mucus layer show a larger coordination length scale, as if the mucus acted as an elastic raft effectively coupling cilia over long distances. This is corroborated by the coordination length scale being larger in samples from patients affected by Cystic Fibrosis than in healthy samples, and much shorter when the mucus layer is washed and cilia therefore beat in a near-Newtonian fluid. We then show how it is possible to employ multiDDM to measure the effectiveness of drugs in recovering, in CF samples, a coordination length scale typical of a healthy phenotype. In the second part I will focus instead on the single cilium scale, showing how we can attempt to link the beating pattern of cilia to numerical simulations studying synchronisation in a model system. In particular in Chapter 5 I will describe our approach to quantitatively describe the beating pattern of single cilia obtained from human airway cells of either healthy individuals or patients affected by Primary Ciliary Dyskinesia. Our description of the beating pattern, and the selection of a few meaningful, summary parameters, are then shown to be accurate enough to discriminate between different mutations within Primary Ciliary Dyskinesia. In Chapter 6 instead I report the results obtained by coarse-graining the ciliary beat pattern into a model system consisting of two ``rotors''. The rotors are simulated colloidal particles driven along closed trajectories while leaving their phase free. In my study, the trajectories followed by the rotors are analytical fits of experimental trajectories of the centre of drag of real cilia. The rotors, that are coupled only via hydrodynamics interactions, are seen to phase-lock, and the shape of the trajectory they are driven along is seen to influence the steady state of the system.

This document describes the design and deployment of a first generation water vapor density sensing unit, the HumiSense. This device is based on an open, air-filled capacitor which is part of a resonant circuit. The frequency of the resonant circuit mixed with a fixed frequency oscillator is the basis of the method to generate a signal that is associated to the change in water vapor density within the open capacitor with time. The physical testing results were inconclusive given that there were many unresolved artifacts in the data. Several suggestions for improving the device for future device generations were provided. / Graduation date: 2013

Over the past several decades, the rate of pulmonary nontuberculous my- cobacterial (PNTM) disease has been increasing. PNTM patients gener- ally consist of lean and tall women presenting with symptoms in the sixth decade of life. They have a de nitive morphophenotype, but no consistent immunological abnormalities despite extensive investigation. I hypothesized that respiratory epithelial dysfunction might play a critical role in PNTM disease predisposition because diseases with defects of mucociliary transport have high rates of PNTM disease that increase with age, suggesting a direct connection between airway epithelial function and PNTM disease. I found that PNTM patients have a distinct respiratory epithelial phenotype ex vivo and decreased nasal nitric oxide levels in vivo. The PNTM ex vivo phenotype consists of an abnormally low resting ciliary beat frequency (CBF) and abnormal CBF response to toll-like receptor (TLR) agonists. The depressed baseline CBF response in PNTM patient cells can be normalized ex vivo by augmenting the nitric oxide-cyclic guanosine monophosphate pathway without appreciable e ect on CBF in healthy controls. In healthy controls, bacterial TLR agonists increase CBF and viral TLR agonists decrease CBF. In PNTM patients these responses are impaired and are not normalized with the normalization of the resting CBF rate. Inhibitor-induced disruption of signalling pathways associated with CBF regulation demonstrated that the majority of the CBF response to TLR agonists involves the PI-3K pathway and PKC. Inhibition of the PI-3K pathway (PI-3K , Akt1, and PDK1) closely mimicked the ex vivo phenotype seen in PNTM patient respiratory epithelia. These data identify a novel aspect of PNTM disease with in vivo and ex vivo correlates that suggest that PNTM infection is associated with abnormal function at both the CBF and TLR response levels. This phenotype is novel, reproducible, and provide a foundation with which to determine the genetic basis of PNTM infection.

Many research studies have analyzed the effect that whole-body vibration (WBV) has on sleep, and some have sought to use vibration to treat sleep disorders such as insomnia. It has been shown that low frequencies (f < 2Hz) are generally sleep inducing, but oscillations of this frequency are typically difficult to achieve using electromagnetic vibration drives. In the research that has been performed, optimal vibration parameters have not been determined, and the effects of multiple vibration sources vibrating at different frequencies to induce a low frequency traveling wave have not been explored. Insomnia affects millions of people worldwide, and non-pharmacological treatment options are limited. A bed excited with multiple vibration sources was used to explore beat frequency vibration as a non-pharmacological treatment for insomnia. A repeated measures design pilot study of 14 participants with mild-moderate insomnia symptom severity was conducted to determine the effects of beat frequency vibration, and traditional standing wave vibration on sleep latency and quality. Participants were monitored using high-density electroencephalography (HD-EEG). Sleep latency was compared between treatment conditions. Trends of a decrease in sleep latency due to beat frequency vibration were found (p ≤ 0.181 for AASM latency, and p ≤ 0.068 for unequivocal sleep latency). Neural complexity during wake, N1, and N2 stages were compared using Multi-Scale Sample Entropy (MSE), which demonstrated significantly lower MSE between wake and N2 stages (p ≤ 0.002). Lower MSE was found in the transition from wake to N1 stage sleep but did not reach significance (p ≤ 0.300). During N2 sleep, beat frequency vibration shows lower MSE than the control session in the left frontoparietal region. This indicates that beat frequency vibration may lead to a decrease of conscious awareness during deeper stages of sleep. Standing wave vibration caused reduced Alpha activity and increased Delta activity during wake. Beat frequency vibration caused increased Delta activity during N2 sleep. These preliminary results suggest that beat frequency vibration may help individuals with insomnia symptoms by decreasing sleep latency, by reducing their conscious awareness, and by increasing sleep drive expression during deeper stages of sleep. Standing wave vibration may be beneficial for decreasing expression of arousal and increasing expression of sleep drive during wake, implying that a dynamic vibration treatment may be beneficial. The application of vibration treatment as part of a heuristic sleep model is discussed.

The thesis is focused on the development of a radioamateur short-wave transceiver. The basic functions, features and parameters are described in the introduction. The bandplan and appropriate types of emission are also included in the introductory part. The frequency synthesis module is discussed in the second part of the document. Emphasis is placed on the direct digital synthesis method (DDS) using modern Analog Devices circuits. The proposed DDS module includes a high-speed clock source. The description of an intermediate frequency module with a demodulator is also placed there. The final part in devoted to the design of a transceiver control panel with a graphical display, a keyboard and a rotary encoder. The firmware for an ATmega128 microcontroller is described in detail at the end of the thesis.

La neteja del moc i dels patògens dels pulmons, i el transport de gàmets i embrions en els òrgans reproductius de les femelles són funcions clau en els epitelis ciliats, tals com aquells que es troben presents en les vies respiratòries i l'oviducte. La taxa de transport mucociliar és funció de la freqüència de batut ciliar (CBF) i aquesta freqüència és augmentada per increments en la concentració de Ca2+ intracelul·lar. El canal catiònic "transient potential vanilloid 4" (TRPV4) intervé en l'entrada de Ca2+ en resposta a estímuls mecànics i osmòtics. L'expressió del TRPV4 en l'epiteli ciliat de les vies respiratòries i de l'oviducte és confirmada mitjançant la localització per immunofluorescència del canal iònic a la membrana apical de l'epiteli ciliat i polaritzat, allà on la senyalització de Ca2+ és requerida per la regulació de la CBF. Cèl·lules ciliades de la tràquea de ratolins TRPV4-/- no expressen el canal TRPV4, no responen a l'activador específic del TRPV4, el 4&#945;-phorbol 12,13-didecanoate (4&#945;-PDD) i presenten respostes de Ca2+ reduïdes a temperatures mitjanes (~25ºC- 8ºC), un altre estímul dels canals TRPV4. L'activació dels canals TRPV4 per solucions altament viscoses i per hypotonicitat depèn de l'activació de la via de la fosfolipasa A2(PLA2)i la subseqüent producció de àcid epoxieicosatrienoic (EET). En condicions de baixa activació de la PLA2, estímuls mecànics i hipotònics alliberen ATP per a l'activació de la via de la fosfolipasa C (PLC)-inositol trifosfat (IP3) per contribuir a l'activació dels canals TRPV4. Descrivim que el metabòlit IP3 sense ser un agonista per ell mateix, sensibilitza el TRPV4 per a l'activació de EET, essent aquest un mecanisme general. L'acoblament funcional entre els canals TRPV4 de la membrana plasmàtica i els receptors de IP3 (IP3R) és necessari tant per iniciar com mantenir la senyalització oscil·latòria del Ca2+ desencadenada per estímuls viscosos i hipotònics. Un dels principals activadors de la CBF, la adenosina-5'-trifosfat (ATP), desencadena una resposta cel·lular mediada per Ca2+ en la que es desencadena tant l'alliberament de Ca2+ des dels dipòsits intracel·lulars com l'entrada de Ca2+. És destacable la contribució de el TRPV4 en l'augment de la CBF mediada per ATP. És més, el nostre treball implica als canals TRPV4 exclusivament en l'entrada de Ca2+ activada per receptor (ROCE). Tot plegat, aquesta tesi doctoral mostra el paper dels canals TRPV4 en l'acoblament d'estímuls fisiològics tipus mecànic, osmòtic i químic a la regulació de la CBF en l'epiteli ciliat destinat al transport mucociliar. / Clearance of mucus and pathogenic agents from lungs and the transport of gametes and embryos in the female reproductive organs are key functions of ciliated epithelia such as those present in the airways and the oviduct. The rate of mucociliary transport is a function of ciliary beat frequency (CBF) and this, in turn, is increased by increases in intracellular calcium. Transient potential vanilloid 4 (TRPV4)cation channel mediates Ca2+ influx in response to mechanical and osmotic stimuli. TRPV4 expression in ciliated epithelia from airways and oviduct is confirmed by immunofluorescence localization of the channel at the apical membrane of the polarized ciliated epithelia, where the Ca2+ signalling is required for CBF regulation. Ciliated tracheal cells from TRPV4-/-mice show no TRPV4 expression, neither increases in intracellular Ca2+ and CBF in response to the TRPV4-specific activator 4&#945;- phorbol 12,13- idecanoate (4&#945;-PDD), and reduced responses to mild temperatures (~25ºC - 38ºC), another TRPV4-activating stimulus. TRPV4 gating by high viscous loads and hypotonicity depends on phospholipase A2 (PLA2) pathway activation and subsequent production of epoxyeicosatrienoic acid (EET). Under conditions of low PLA2 activation, mechanical and hypotonic stimuli use extracellular ATP release-mediated activation of phospholipase C (PLC)-inositol triphosphate(IP3)signalling to support TRPV4 gating. We describe that IP3, without being an agonist itself, sensitizes TRPV4 to EET activation. Besides, the functional coupling between plasma membrane TRPV4 channels and IP3 receptors (IP3R) is required to initiate and maintain the cellular oscillatory Ca2+ signal triggered by high viscous loads and hypotonic stimuli. One of the main CBF activators, adenosine-5'-triphosphate (ATP), triggers both Ca2+ release from intracellular Ca2+ stores and Ca2+ entry. Interestingly, TRPV4 contributes to ATP-induced increase in CBF. Furthermore, our work implicates TRPV4 channel exclusively in receptor-operated Ca2+ entry. Collectively, this PhD thesis shows the role of TRPV4 channels coupling physiologically relevant mechanical, hypotonic and chemical stimuli to CBF regulation in motile ciliary epithelia.