This dissertation involved a theoretical and experimental investigation of the adhesive forces between spherical particles of four different diameters and two selected flooring materials under different air velocities. Previous theoretical work and experiments described in the literature tended to be conducted with idealized surfaces, and therefore have limited applicability to indoor environments. Controlled experiments were designed, constructed and executed to measure the air velocity required to overcome adhesion forces. The diameters of the particles investigated were 0.5, 3.0, 5.0 and 9.9 [mu]m, and the flooring materials were linoleum and wooden flooring. The critical velocity, the flow at which 50% of the particles detached, is presented as a function of particle diameter for each surface. The measured values were then compared to empirical and theoretical models as well as to a scaling analysis that considers component forces that act on a particle-surface system. The results suggest that critical velocity decreases with increasing particle diameter and that existing models have limited applicability to resuspension from flooring materials. / text

A Bacteroides gingivalis adhesin mediating attachment to red blood cells and buccal epithelial cells was isolated, cloned and characterized. The isolation procedure involved gentle stirring of the cells followed by ammonium sulphate precipitation, ion-exchange and gel chromatography. The native molecule had a Mr in excess of 10⁶ kDa and was made up of subunits with an Mr of 43 kDa. Antisera raised to the adhesin and its subunits reacted with antigens on the surface of B. gingivalis cells. No reaction with fimbriae was seen. The IgG fractions from these antisera inhibited the adherence of B. gingivalis to host tissue. Proteolytic enzymes destroyed binding capability of whole cells and of the purified adhesin but the molecular weight of the haemagglutinin was not altered. A genomic library of B. gingivalis DNA was created in E. coli JM83. 5500 colonies were screened by a colony immunoassay with anti-S. gingivalis serum and by a direct haemagglutinating assay. 337 clones tested positive by the immunoassay and two clones, 1-3,and 1-49 tested positive for haemagglutinating activity. Both haemagglutinating positive recombinants had inserts of 3.2 kb. One clone, 1-49 was chosen for further characterization. E. coli 1-49 expressed a protein of 43 kDa that was not present in E. coli JM 83 control as seen by SDS-PAGE and Western blot analysis. Anti-1-49 serum inhibited the haemagglutinating activity of B. gingivalis and E. coli 1-49. This serum reacted with surface molecules on B. gingivalis and E. coli 1-49 as seen by immunogold electron microscopy and immunofluorescence, and to the purified haemagglutinin by Western blot analysis. Like the haemagglutinin on B. gingivalis, the haemagglutinating activity of E. coli 1-49 was destroyed by heating and proteolytic enzymes but the apparent size of the molecule as determined by SDS-PAGE was not affected. A bacterial coaggregating adhesin from B. gingivalis was isolated and characterized. The isolation procedure involved adsorption of the solubilized adhesin on S. mitis followed by elution with glycine buffer. SDS-PAGE of the boiled adhesin revealed a protein with an Mr of 46 kDa. Proteolytic digestion destroyed all bacterial aggregating activity and hydrolysed the 46 kd protein. Antisera raised to the 46 kDa protein reacted with surface molecules on all strains of B. gingivalis tested. This antiserum inhibited the coaggregation reaction between B. gingivalis and other bacteria. Vesicles produced by B. gingivalis were found to enhance the binding of S. sanguis to serum coated hydroxy apatite (SeHA). Maximum vesicle mediated binding took place at 37°C and was destroyed by heating. The lipopolysaccharide from several black pigmented bacteroides were isolated and characterized physically, chemically and immunologically. All of the LPS were of the smooth type and contained the sugars rhamnose, glucose, galactose, glucosamine and galactosamine; no KDO or heptose were found. / Science, Faculty of / Microbiology and Immunology, Department of / Graduate

L'ICAP-1 est impliqué dans la dynamique de l'intégrine et la génération de force en contrôlant l'endocytose de l'intégrine grâce à la scission dépendant de nm23 des puits endocycliques recouverts de chlatrine.L'ICAP-1 a été identifié comme un partenaire spécifique de l'intégrine b1 (Degani et al., 2002, Zhang et Hemler, 1999). Nous avons déjà montré que l'ICAP-1 est impliqué dans la réponse mécanisée aux cellules et la différenciation cellulaire d'une manière dépendante de l'intégrine b1 (Bouvard et al., 2007; Brunner et al., 2011; Faurobert et al., 2013; Millon-Frémillon et al. , 2008; Renz et al., 2015). Cependant, comme ICAP-1 est également capable d'adapter la migration cellulaire en réponse à la rigidité du substrat d'une manière indépendante de la β1-intégrine (Bouin et al., 2017), nous avons spéculé sur un rôle plus général de l'ICAP-1 dans l'adhésion cellulaire et dynamique d'adhérence focale. Pour cela, nous avons créé un environnement cellulaire où l'intégrine b1 et / ou l'ICAP-1 étaient absentes en utilisant quatre lignées cellulaires: les ostéoblastes WT, les cellules ostéoblastes KO de l'intégrine b1, les cellules ostéoblastes KAP ICAP-1 et les cellules ostéoblastes double KO b1 / ICAP-1 afin de surveiller le comportement de l'intégrine b3. Comme prévu, l'épuisement de l'intégrine b1 est associé à la perte d'étalement cellulaire et à la génération de force selon la mesure de la microscopie par force de traction. De manière surprenante, la suppression supplémentaire de ICAP-1 (b1 intégrine et ICAP-1 KO) conduit à la restauration de l'étalement cellulaire et de la génération de force qui dépend de l'intégrine b3. Ces forces médiées par l'intégrine b3 sont corrélées avec la diffusion lente de l'intégrine b3 dans les sites d'adhésion et le renouvellement lent de l'adhésion focale contenant l'intégrine b3 (FRAP / TIRF / vidéomicroscopie). Nous avons abordé la question de savoir si ICAP-1 pourrait réguler l'endocytose de l'intégrine b3 puisque ICAP-1 interagit avec nm23-H2 (Fournier et al., 2002), un nucléoside diphosphate kinases (NDPK) impliqué dans l'endocytose à médiation par dynamine en produisant du GTP à travers l'adénosine triphosphate (ATP) de conversion du diphosphate de guanosine (PIB) (Boissan et al., 2014). Nous montrons que la suppression de nm23 ou de dynamine ou de chlatrine dans les cellules épuisées dans l'intégrine b1 est capable d'imiter la perte combinée de l'intégrine b1 et de l'ICAP-1 en rétablissant l'étalement cellulaire, la génération de force et la dynamique de l'intégrine b3. Pour confirmer l'implication de l'ICAP-1 dans l'endocytose de l'intégrine b3, nous montrons que l'absorption de l'anticorps de l'intégrine b3 est efficacement bloquée dans les cellules épuisées dans ICAP-1. Nos résultats suggèrent que ICAP-1 pourrait être impliqué dans la dynamique de l'intégrine et la génération de force en contrôlant l'endocytose de l'intégrine grâce à la scission dépendant de nm23 des puits endocytaires de la couche de chlatrine. / ICAP-1 is involved in integrin dynamics and force generation by controlling integrin endocytosis through nm23-dependent scission of endocytic chlatrin coated pits.ICAP-1 has been identified as a specific partner of b1 integrin (Degani et al., 2002; Zhang and Hemler, 1999). We have previously shown that ICAP-1 is involved in cell mechanoresponse and cell differentiation in a b1 integrin dependent manner (Bouvard et al., 2007; Brunner et al., 2011; Faurobert et al., 2013; Millon-Frémillon et al., 2008; Renz et al., 2015). However, as ICAP-1 is also able to adapt cell migration in response to substrate stiffness in a β1-integrin-independent manner (Bouin et al., 2017), we speculated on a more general role of ICAP-1 in cell adhesion and focal adhesion dynamics. For this purpose we have created cellular environment where b1 integrin and/or ICAP-1 were absent by using four cell lines: WT osteoblast, b1 integrin KO osteoblast cells, ICAP-1 KO osteoblast cells and double KO b1/ICAP-1 osteoblast cells in order to monitor b3 integrin behavior. As expected, depletion of b1 integrin is associated with the loss of cell spreading and force generation according traction force microscopy measurement. Surprisingly, the supplementary deletion of ICAP-1 (b1 integrin and ICAP-1 KO) leads to restoration of cell spreading and force generation which are dependent on b3 integrin. These b3 integrin-mediated forces are correlated with slow diffusion of b3 integrin within adhesion sites and slow turnover of b3 integrin containing focal adhesion (FRAP/TIRF/videomicroscopy). We addressed the question whether ICAP-1 might regulate b3 integrin endocytosis since ICAP-1 interacts with nm23-H2 (Fournier et al., 2002), a nucleoside diphosphate kinases (NDPKs) involved in dynamin-mediated endocytosis by producing GTP through adenosine triphosphate (ATP)–driven conversion of guanosine diphosphate (GDP) (Boissan et al., 2014). We show that the deletion of either nm23 or dynamin or chlatrin in cells depleted in b1 integrin is able to mimic the combined loss of b1 integrin and ICAP-1 by restoring cell spreading, force generation and b3 integrin dynamics. To confirm the involvement of ICAP-1 in b3 integrin endocytosis, we show that the b3 integrin antibody uptake is efficiently blocked in cells depleted in ICAP-1. Our results suggest that ICAP-1 might be involved in integrin dynamics and force generation by controlling integrin endocytosis through nm23-dependent scission of endocytic chlatrin coated pits.

In this thesis, a direct method of Atomic Force Microscopy (AFM) technique has been developed to measure the adhesion forces between BSA and two different surfaces: mica (a hydrophilic surface); and polystyrene (a hydrophobic surface); in PBS solution. We have shown possible to measure interactions between proteins and substrate surface directly without any modification to the substrate and the AFM tip; this means protein molecules can keep the natural elastic property within the force measurements. The average measured value of adhesion forces between BSA and mica is 0.036 ± 0.002 nN, and between BSA and polystyrene is 0.066 ± 0.003 nN. The polystyrene surface is more adhesive to BSA than the mica surface. This is consistent with previous research, which assessed that hydrophobic surfaces enhance protein adhesion but hydrophilic surfaces do not.

No description available.

The work presented in thesis is part of the DANIELA project which aims to replace the current air data system on civil aircrafts with a flush mounted Air Data System (ADS) built around a 3 axis Doppler LIDAR function as the primary data channel. This thesis is focused on the comparison of different window materials and their optical clarity by means of theoretical and experimental analysis. Four different window materials including BK7, Sapphire, Germanium and ZnS are placed in a wind tunnel. The samples are each exposed to flows of air and water for specific periods of time during which temperature, pressure and air speed are recorded. Subsequently, each sample is carefully observed under the microscope. This is followed by the measurement of the amount of back scatter via detecting the change in the voltage once it is placed in the optical station. The optical tests reveal the amount of dust adhered to the samples which results in increased voltage. Review of these samples under the microscope matches the results obtained from the optical test. The two sets of data obtained from the two tests determined that some samples collected more dust in comparison to others. It was established that under identical test conditions i.e. flow, temperature and moisture, BK7 and Sapphire collect considerably less dust compared to ZnS. Moreover it was impossible to test Germanium sample optically, under a microscope as it is a dark opaque glass.

published_or_final_version / Biochemistry / Doctoral / Doctor of Philosophy

published_or_final_version / Molecular Biology / Master / Master of Philosophy

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mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:"Times New Roman"; mso-bidi-theme-font:minor-bidi;} --><!--[endif]--></p><p>The aim of the project was to develop a method to investigate differences in adhesion of microorganisms to materials that contains lignocellulose. The method was tested on a gram-positive (<em>Micrococcus lutea) </em>and one gram-negative (<em>E-coliJM109</em>) bacteria.</p><p>The study was begun by cultivation of the two microorganisms. The cultivation was done to calculate the generation times of the bacteria and to obtain growth curves. Cells from these cultivations were also frozen (-70ºC) and later used for inoculation.</p><p>At STFI-Packforsk AB the total charge on the mass was measured and later a conductivity titration on the mass was executed as well, all to find out more about the different properties of the mass. Properties that in a later part of this study could possibly be linked to the adhesion of cells to the pulp. The adhesion experiments that were executed gave poor results. The adhesion experiment with <em>M. lutea</em> was the only experiment that gave a reproducible result. In this experiment <em>M. lutea </em>was contacted with bleached leaf. A reduction of cells was observed in all of the dilutions where <em>M. lutea</em> had been in contact with the mass. The number of colony forming units of the culture was 1,2×10⁷ before the adhesion and 2×10⁶ subsequently.</p><p> </p>

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