The central theme of the thesis is the current conceptualization of social classes and the issue of stratification in the Czech Republic in the period after 1989. In the first part I confront key theses of classical stratification concepts and follow their transformations in the history of exploration of the social structure. In the second part, by secondary analysis of quantitative data, I am trying to find the key factors of self- classification into social structures and to determine how this subjective self- classification correlates with objective (sociological) classification. An important part of the research is also the question of to what extent and in what sense people perceive the contemporary Czech society as a social-stratified. In the conclusion, on the basis of the results of analysis, I am trying to suggest which of the classical concepts of stratification best reflects the situation in contemporary Czech society. Key words: subjective and objective position, stratification, class, stratum, status, society, social structure

Charles University, Faculty of Pharmacy in Hradec Králové Department of Biophysics and Physical Chemistry Author: Lucia Dulanská Supervisor: Mgr. Petra Pullmannová, Ph.D Title of thesis: Optimization of the skin barrier model with isolated ceramides of human Stratum corneum Stratum corneum (SC), the uppermost layer of the skin, regulates transcutaneous water loss and protects against outer conditions and harmful substances. It consists of cornified cells - corneocytes and extracellular lipid matrix, which is responsible for the barrier functions. Corneocytes are covered with covalently bound lipids creating the corneocyte lipid envelope (CLE). CLE is considered to interconnect the extracellular lipids with corneocytes and to have a templating effect. We aimed to optimize a skin lipid model simulating also the presence of CLE. The lipidic part of the model was prepared from an equimolar mixture of isolated human skin ceramides (hCer), cholesterol and free fatty acids (FFA, either protonated or deuterated) with 5 weight % of cholesteryl sulfate. hCer were extracted from the isolated human SC and purified by the column chromatography. The composition of hCer was determined by the high- performance thin-layer chromatography. The reverse-phase and normal phase silica gel particles served as the CLE...

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal. / Une étude a été entreprise dans le but de déterminer les interactions pouvant survenir entre certains lipides impliqués dans les fonctions de barrière de perméabilité et de desquamation du stratum corneum. Cette étude a consisté à étudier le thermotropisme de diverses dispersions composées de sphingomyéline, céramide 3, acide palmitique perdeutéré, cholestérol et sulfate de cholestérol en proportions variées, par spectroscopie infrarouge par transformée de Fourier. Cette étude a été également effectuée aux pH 5.2 et 7.4 afin d'estimer l'influence de l'environnement ionique sur 1'interaction de ces lipides. Le thermotropisme des dispersions de céramide 3 et de sphingomyéline a indiqué que le céramide 3 adopte un empilement plus serré que la sphingomyéline aux deux pH. Les résultats ont également indiqué que l'acide palmitique rigidifie la bicouche de sphingomyéline et fluidifie la dispersion de céramide 3, et que la miscibilité de cet acide gras est plus élevée avec la sphingomyéline qu'avec le céramide 3. Toutefois, la miscibilité de l'acide palmitique et du céramide 3 est augmentée par le cholestérol et le sulfate de cholestérol à pH 5.2, ce qui semble affecter leur thermotropisme en induisant la formation d'une phase très ordonnée à 37 °C. Par contre, le cholestérol et le sulfate de cholestérol semblent accentuer la déprotonation de l'acide palmitique en présence du céramide 3 à pH 7.4, ce qui semble provoquer une séparation de phase de l'acide gras et du céramide 3. Le cholestérol induit également la formation de la phase liquide ordonnée au sein de la bicouche de sphingomyéline en présence de l' acide palmitique aux pH 5.2 et 7.4. Toutefois, le sulfate de cholestérol semble être moins efficace que le cholestérol à abolir la transition de phase de la sphingomyéline et de l 'acide palmitique. L'analyse des résultats nous a permis de déterminer que l'acide palmitique, le cholestérol et le sulfate de cholestérol interagissent avec la sphingomyéline et le céramide 3 par l'intermédiaire d' interactions hydrophobes et de van der Waals impliquant leurs portions hydrophobes, et de liaisons hydrogène qui impliquent leurs groupements d'interface et de tête iv polaire. Il ressort également que les implications de ces lipides et du pH dans les fonctions de barrière de perméabilité et de desquamation pourraient être purement structurales. En fait, la structure de la tête polaire du céramide 3 permet l'empilement serré des lipides ainsi que l'imperméabilité des membranes, le cholestérol et le sulfate de cholestérol augmentent la miscibilité de l'acide palmitique et du céramide 3 à pH 5.2, et l'environnement à pH 5.2 semble être nécessaire au maintien de l'intégrité du stratum corneum.

No / This work explored the role of skin appendages (shunt route) in passive and iontophoretic drug and liposome penetration. The technique used an epidermis and stratum corneum sandwich from the same skin donor with the additional stratum corneum forming the top layer of the sandwich. Penetration was monitored during occluded passive and iontophoretic (0.5 mA cm-2) delivery of mannitol and estradiol solutions, and ultradeformable liposomes containing estradiol. The shunt route had a significant role during passive penetration of mannitol (hydrophilic compound), but was negligible during penetration of estradiol (lipophilic drug) and liposomes. In iontophoresis, the shunt route significantly contributed to the overall flux of all preparations, being highest for mannitol. However, shunts were not the only pathway for iontophoretic drug delivery and evidence was observed for the creation of new aqueous pathways via disorganization of the intercellular lipid domain of stratum corneum. The skin sandwich technique should prove valuable for general studies on routes of skin penetration.

Zielsetzung dieser Arbeit war es, das Verhalten der in modernen Sonnenschutzmitteln eingesetzten Titandioxid-Mikropartikel mit quantitativen Methoden zu bestimmen, um Aussagen über ihre Eignung zu erhalten. Dabei stand die Frage nach der Verteilung der Substanz innerhalb des Stratum corneum im Mittelpunkt der Untersuchungen. Insbesondere war wegen der bekannten photokatalytischen Aktivität von Titandioxid zu klären, ob ein Kontakt mit den lebenden Bereichen der Haut ausgeschlossen werden kann. Für die Messungen wurden zwei kommerziell genutzte, unterschiedlich beschichtete Titandioxid-Mikropartikel eingesetzt. Eine wesentliche Voraussetzung für diese Untersuchungen war die Anwendung der Abrissmethode (Tape stripping) in Kombination mit der spektroskopischen Bestimmung der Extinktion im sichtbaren Bereich zur Berechnung des Hornschichtprofils. Die Konzentration der Titandioxid-Partikel wurde mit Hilfe von Röntgenfluoreszenz-Messungen bestimmt. So war es erstmals möglich, in vivo standardisierte und reproduzierbare Untersuchungen zum Penetrationsverhalten von beschichteten Titandioxid-Mikropartikeln in die Hornschicht der menschlichen Haut durchzuführen. Durch Langzeitapplikation der Substanzen und die Beobachtung der Titandioxid-konzentrationen in der Hornschicht über mehrere Tage konnten auch Aussagen zum Penetrationsverhalten der applizierten Mikropartikel gemacht werden. Es wurde eindeutig gezeigt, dass die untersuchten Substanzen unabhängig von ihrer Struktur, von ihrer Beschichtung und vom Probanden hauptsächlich in den obersten Schichten des Stratum corneum lokalisiert sind. Nach Klärung dieser grundsätzlichen Fragen war es notwendig, die Ursache für das Auftreten extrem geringer Titandioxid-Konzentrationen auf Abrissen zu bestimmen, die aus tieferen Schichten des Stratum corneum entnommen wurden. Durch die Kombination der Abrissmethode mit einem speziellen Färbeverfahren und der Laser-Scan-Mikroskopie ergaben sich deutliche Hinweise auf die Bedeutung der Follikelkanäle für das beobachtete Phänomen. Röntgenspektroskopische Untersuchungen an Biopsien zeigten, dass diese Mikropartikel in die Haarfollikel eindringen und damit Bereiche unterhalb des Stratum corneum erreichen. Hierbei wurden die Mikropartikel in dieser Region nur in Follikelkanälen, nicht aber im Bereich der lebenden Zellen nachgewiesen. Diese Ergebnisse belegen, dass Titandioxid nur in einzelne Follikelkanäle penetriert, eine Aussage, die im Rahmen der vorliegenden Untersuchungen erstmals beschrieben wurde. In Übereinstimmung mit der Zielsetzung der Arbeit konnte unter Einsatz unterschiedlicher Untersuchungsmethoden gezeigt werden, dass die beschichteten Titandioxid-Mikropartikel im oberen Bereich der Hornschicht lokalisiert sind und damit als hocheffiziente Lichtschutzfilter den Schutz der darunter liegenden lebenden Bereiche der Haut garantieren. Der sichere Nachweis der Titandioxid-Mikropartikel innerhalb einzelner Follikelkanäle besitzt grundsätzliche Bedeutung für das Verständnis von Penetrationswegen. / The objective of this work was to determine the behaviour of titanium dioxide microparticles, used in modern sunscreen agents, with quantitative methods in order to receive statements about their suitability. The focal point of the investigations was the question on the distribution of the substance within the stratum corneum. Because of the well-known photocatalytic activity of titanium dioxide, our aim was to clarify whether a contact with the living cells of the skin could be excluded. For these measurements, two commercial titanium dioxide micro particles were used, covered with different coating materials. The calculation of the stratum corneum profile was a substantial prerequisite for these investigations. This was done by application of the tape stripping method in combination with the spectroscopic determination of the absorption in the visible range. The concentration of the titanium dioxide particles on the tape strips was determined by X-ray fluorescence measurements. In this manner it was possible for the first time, to perform standardized and reproducible in vivo measurements to investigate the penetration behaviour of coated titanium dioxide micro particles into the horny layer of the human skin. Predications could be made about the penetration behaviour of the applied micro particles by observation of the titanium dioxide concentrations in the stratum corneum during several days after long-term application of the substances. It could be shown that the examined substances were localized mainly in the upper layers of the stratum corneum, independently of their structure, coating and the volunteers. After clarifying these basic questions, it was necessary to explain the occurrence of extremely small concentrations of titanium-dioxide on tape strips, which were taken from deeper layers of the stratum corneum. This observed phenomenon was investigated by combining tape stripping with a selective staining of the tapes and with laser scanning microscopy. Investigations of a biopsy with x ray fluorescence measurements showed that the micro particles penetrate into the hair follicles, and in this manner reached areas below the stratum corneum. In this region, the micro particles were found only in follicle channels, but not within the area of the living cells. These results prove that titanium dioxide micro particles only penetrate into single follicle channels, a statement, which has been described here for the first time. In agreement with the objective of this thesis, it was shown, that the coated titanium dioxide micro particles were localized within the upper area of the horny layer and as high efficient sunscreen filters consequently guarantee, the protection of the living areas of the skin below. The proof of the titanium dioxide micro particles within individual follicle channels has basic importance for the understanding of the penetration pathways.

Energy use in buildings account for about one third of the total global energy supply and contributes as much as 30% of the anthropogenic greenhouse gas emissions. It is estimated that energy use in buildings will increase to 67% by 2030. The need for better thermal comfort and air quality in indoor environments is the leading cause for high energy use in buildings.  Heating, ventilation and air conditioning systems take up about 50% of the total energy use in buildings which is about 10-20% of the national energy use in most developed countries. The development and adoption of sustainable ventilation systems is a viable solution to mitigate climate change and curtail carbon emissions. The experimental study was conducted in a room resembling a modern office in a laboratory environment. The study involved investigating the ability of the system to provide cooling and heating. Concentration decay tracer gas technique using Sulphur hexafluoride (SF6) gas was used to determine the local air change index and air change efficiency in the room. Low-velocity omni-directional thermistor anemometer type CTA88 were used to measure the air velocity and temperature in the room. Smoke was used to visualise the flow patterns created in the room.  The climate chamber was used to mimic climatic conditions in winter. Fifteen cases were investigated with five air flow rates set points (30, 40, 50, 60 and 70 l/s) at three supply air temperatures, i.e., 17.6 °C, 21.0 °C and 25.3 °C. The results of the local air change index and air change efficiency for the nominal supply temperature of 17.6 °C showed that the system had strong characteristics of a mixing ventilation system. At the supply air temperature of 21.0 °C, the performance of the system deteriorated slightly to below that of a mixing ventilation system and could not satisfactorily provide heating at supply temperature of 25.3 °C. Better performance of the system at all supply air temperature setpoints was observed at lower airflow rates. At all supply air temperature setpoints, relatively higher degree of temperature stratification was observed at lower supply. The draught rate levels decreased with increase in supply air temperature and height. The location of the air inlet terminals in relation to the workstations had significant effect on the performance of the system. The stratum ventilation system did not work efficiently because the air streams were heavily mixed before reaching the occupants.

Skin, more specifically the outermost skin layer, the stratum corneum (SC), forms an extremely effective barrier, preventing both the loss of heat and water, and the ingress of micro-organisms and chemicals. Assessing the rate and extent of drug permeation into or through the skin is important both to evaluate the usefulness of a drug for topical or transdermal delivery, and to compare different formulations to assess their bioequivalence. Prediction of drug permeation is logistically, ethically and economically preferable to in vivo measurements. The recent progress that has been made with empirical and mechanistic mathematical models, along with in vitro diffusion cell techniques has been reviewed. However, currently, in vivo measurements, in man, are still required. For new chemical entities, the need for clinical trials is clear. In the case of generic products, however, there is considerable effort currently being expended to replace expensive, subjective clinical trials with objective, validated measurements of drug permeation, in vivo, in particular to assess bioequivalence. The tape stripping technique has emerged as a promising technique to objectively measure drug permeation through skin, and is the focus of this thesis. After formulation application and removal, layers of SC are sequentially removed by adhesive tapes. As the SC performs the main barrier function of the skin, measuring the rate and extent of drug permeation through this layer is assumed to be related to overall topical bioavailability. The work in this thesis concentrates on performing tape stripping studies such that all tapes are analysed individually, and drug concentration as a function of SC depth is measured. The concentration depth profiles across the SC may be fitted to an appropriate solution of Fick's second law of diffusion to obtain estimates of the vehicle-SC partition coefficient and the drug's diffusivity in the membrane. These dermato-pharmacokinetic parameters may be compared for different formulations.

Atopic eczema is an incurable disorder of the skin. Sufferers are afflicted with hypersensitivity to environmental agents such as soaps (detergents), animal dander, pollen, specific foods and sometimes even water. Genetic mutations in atopic eczema compromise the development of the stratum corneum resulting in xerotic skin that is prone to cracking and increased permeability which leads to irritation due to the influx of exogenous material through the skin. The causes of atopic eczema are due to a combination of genetic and environmental factors and it is, therefore, a difficult disease to manage. Emollients and topical corticosteroids are the mainstay treatments for eczema. However, they do not treat the underlying cause of the flare-ups frequently seen in the condition; the damaged skin barrier. Defects in the skin barrier arise from premature desquamation of the stratum corneum. The main contributors to barrier breakdown are the up-regulation of skin proteases that are located in the skin. Since zinc is a known protease inhibitor, it would thus follow that a topical treatment for skin barrier repair should be developed. Therefore, the main objectives of this thesis are to successfully incorporate zinc into a formulation to develop a novel class of treatment for eczema and to assess the delivery of the element into the skin. In this thesis, methods to assess and characterise changes to skin barrier function and to extract and quantify zinc in the stratum corneum have been established. The development of two novel topically applied formulations containing zinc lactate as the active ingredient (1% w/w zinc lactate cream and a 2% w/v zinc lactate formulation) has been achieved and the uptake of zinc from the preparations in vitro determined. Further, the in vitro percutaneous penetration of zinc from three commercially available preparations has been investigated and compared to that recovered from the stratum corneum after passive diffusion with the novel zinc formulations. Additionally, in vivo uptake of zinc into human stratum corneum from Sudocremis ® reported. Scanning electron microscopy has revealed the distribution of zinc on the surface of skin treated with various formulations and has also allowed the efficiency of two cleaning procedures to be ascertained. The delivery of zinc from the novel topical formulation; 1% w/w zinc lactate cream, was more efficient than that the three commercial formulations and shows promise as a new approach to treat atopic eczema.

A pesquisa tem por objetivo, analisar as influências da urbanização concentrada (compacta), e de como essa característica altera as condições ambientais e o equilíbrio das formas de relevo local , modificando o estrato geográfico da paisagem. Para essa análise, t i vemos como supor te o mapeamento da evolução do uso da terra urbana em Taboão da Serra em dois momentos distintos, no período compreendido entre 1962 e 2007. O uso da terra tem contornos distintos no contexto intra-urbano, fato este que é traduzido pela evolução e pelo tipo de uso e ocupação dado à paisagem urbana em Taboão da Serra. Os processos geoecológicos encontram- se modificados na sua gênese e na sua estrutura, o que resulta em formas mais antropizadas do que em décadas passadas. A cidade de Taboão da Serra, é a sexta cidade mais compacta do Brasil , fato este que tem reflexos negativos sobre os processos naturais onde, a morfogênese é dominante por conta do intenso uso da terra seja ele para fins imobiliários como também pelo processo de intensificação da cobertura da superfície por materiais que interrompem a percolação das águas pluviais associado à retirada da cobertura vegetal . A escolha da cidade como área de estudo deu-se pelos motivos explicitados acima mas também, pelo fato de que através da comparação de dois momentos da ocupação urbana (1962 e 2007) que são caracterizadas pela intensificação do uso da terra (utilizando fotografias aéreas ) , é possível mapear e representar como o tipo de uso e ocupação pode comprometer em maior ou menor grau as condições geoecológicas de um geossistema. As unidades de paisagem urbana serão compartimentadas tendo como final idade a comparação para diagnosticar a relação atual entre relevo/ solo e uso da terra nesta c idade, que serão representadas na carta de Uso da Terra. / The research aims to examine the influences of urbanization concentrated (compact) , and how this feature changes the environmental conditions and the balance of local landforms by modi fying the geographic stratum of the landscape. For this analysis, we had to support the mapping of the evolution of urban land use in Taboão da Serra at two different times in the period between 1962 and 2007 . The use of land has different contours in the intra-urban context , a fact that is translated by the change in type of use and occupation since the urban landscape in Taboão da Serra. Geoecological processes are modified in their genesis and struc ture, resulting in forms more disturbed than in decades past . The city of Taboao da Serra, is the sixth most compact city of Brazil , a fact that has negative effects on natural processes where the dominant morphogenesis is due to the intense use of land be it for housing but also by the process of intensification of surface coverage by materials that disrupt the percolation of rainwater associated with the removal of the canopy. The choice of the city as the study area took place for the reasons explained above but also by the fact that by comparing two moments of urban occupation (1962 and 2007) which are characterized by the intensification of land use (using aerial photographs) it is possible to map and represent the type of use and occupation can affect a greater or lesser degree the conditions of a geoecological geosystem. The units of the urban lands cape will be compartmentalized view to the comparison to diagnose the current relationship between topography/soil and land use in this city, which will be represented in the land use map.

The world-wide transdermal drug delivery market is quite large, but only a small number of agents have FDA approval. The primary reason for such limited development is the difficulty in permeating the stratum corneum layer of human skin. In our study, we developed a novel percutaneous delivery enhancing approach. Magainin peptide was previously shown to disrupt vesicles from stratum corneum lipid components and this ability of magainin allows us to propose that magainin can increase skin permeability. Therefore, we tested the hypothesis that magainin, a pore-forming peptide, can increase skin permeability by disrupting stratum corneum lipid structure and that magainin¡¯s enhancement requires co-administration of a surfactant chemical enhancer to increase magainin penetration into the skin. In support of these hypotheses, synergistic enhancement of transdermal permeation can be observed with magainin peptide in combination of N-lauroyl sarcosine (NLS) in 50% ethanol-PBS solution. The exposure to NLS in 50% ethanol solution increased in vitro skin permeability to fluorescein 15 fold and the addition of magainin synergistically increased skin permeability 47 fold. In contrast, skin permeability was unaffected by exposure to magainin without co-enhancement by NLS-ethanol. To elucidate the mechanism of this synergistic effect, several characterization methods such as differential scanning calorimetry, Fourier transform infrared spectroscopy, and X-ray diffraction were applied. These analyses showed that NLS-ethanol disrupted stratum corneum lipid structure and that the combination of magainin and NLS-ethanol disrupted stratum corneum lipids even further. Furthermore, confocal microscopy showed that magainin in the presence of NLS-ethanol penetrated deeply and extensively into stratum corneum, whereas magainin alone penetrated poorly into the skin. Together, these data suggest that NLS-ethanol increased magainin penetration into stratum corneum, which further increased stratum corneum lipid disruption and skin permeability. Finally, skin permeability was enhanced by changing the charge of magainin peptide via pH change. We modulated pH from 5 to 11 to change the magainin charge from positive to neutral, which decreased skin permeability to a negatively charged fluorescein and increased skin permeability to a positively charged granisetron. This suggests that an attractive interaction between the drug and magainin peptide improves transdermal flux.