Rozprawy doktorskie: „Bacterial studies”

1

Leigh, J. A. "Studies on bacterial dehalogenases". Thesis, Nottingham Trent University, 1986. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.376092.

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2

Stenbäck, Anders. "Studies of Experimental Bacterial Translocation". Doctoral thesis, Uppsala University, Department of Surgical Sciences, 2005. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-5933.

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One of the main obstacles to maintaining patients with short bowel syndrome on parenteral nutrition, or successfully transplanting these patients with a small bowel graft, is the many severe infections that occur. Evidence is accumulating that translocating bacteria from the patient’s bowel causes a significant part of these infections. In this thesis bacterial translocation is studied in a Thiry-Vella loop of defunctionalised small bowel in the rat.

Bacterial translocation to the mesenteric lymph nodes (MLNs) occurs in almost 100% of the rats after three days. No systemic spread of bacteria is observed unless there is additional immunosupression with depletion of Kupffer cells in the liver. However, blocking the function of α/β T cells does not increase the translocation. Removal of MLNs does not either aggravate bacterial translocation in the Thiry-Vella loop model. Conversely, after small bowel transplantation translocating bacteria spread systemically if the MLNs are removed.

The Thiry-Vella loop should also be a suitable model for the testing of potentially translocation-inhibiting substances. Reinforcement of the intestinal barrier with glutamine or phosphatidylcholine proved insufficient in decreasing bacterial translocation. Even selective bowel decontamination with tobramycin failed to abolish bacterial translocation. Thus, it seems that the driving force for translocation in this model is strong regardless of the relatively small trauma of intestinal defunctionalisation.

Flow cytometric studies of the immune cells in the spleen MLNs showed a decrease in MHC class II positive T cells in the MLNs of the Thiry-Vella loop. Concurrently the number of macrophages increased with time as observed by immunohistochemistry. The fraction of MHC class II negative macrophages increased in the spleens of rats treated with glutamine.

In conclusion, the Thiry-Vella loop model offers possibilities of immunological as well as mechanistic studies on bacterial translocation from small intestine.

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3

Chiu, Cecilia P. C. "Crystallographic studies of bacterial sialyltransferases". Thesis, University of British Columbia, 2007. http://hdl.handle.net/2429/31274.

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Sialic acids terminate oligosaccharide chains on the surfaces of mammalian cells and many microbial species, often playing critical biological roles in recognition and adherence. The enzymes which transfer the sialic acid moiety to these key terminal positions are known as sialyltransferases. Despite their important biological roles very little is understood about the mechanism of action or molecular structure of these enzymes. Campylobacter jejuni, a highly prevalent food-borne pathogen that causes acute gastroenteritis in humans, contains two versions of a sialyltransferase: a monofunctional α-2,3-sialyltransferase Cst-I and a bifunctional α-2,3/8-sialyltransferase Cst-II. Both of these enzymes are responsible for lipooligosaccharides (LOS) sialylation to camouflage the bacterial surface from the host, and thus evade the immune system. In addition, sialylated-glycoconjugates on C. jejuni often mimic human gangliosides, contributing to the molecular basis of Guillain-Barré syndrome, an autoimmune disease of the peripheral nervous system that often develops post-infection. The sialyltransferase reaction is believed to proceed through an inversion mechanism, catalyzing the transfer of sialic acid from CMP-N-acetylneuraminic acid onto different acceptors. This thesis is to understand through high-resolution structural characterization, site specific mutagenesis and kinetic analysis, the mechanism of the glycosyl transfer(s) in both monofunctional and bifunctional Csts. Crystals of Cst-II were obtained and the complex structures with bound CMP, inert donor sugar analogue CMP-3-fluoro-N-acetylneuraminic acid (CMP-3FNeu5Ac) and inhibitor CDP were solved using MAD phasing from incorporated selenomethionines. Work within this study represents the first known structure of a sialyltransferase. Based on the position of the substrates, the active site of Cst-II has been elucidated. Site-directed mutagenesis of conserved residues in the active site was performed and mutants were characterized using enzyme kinetics. A reaction mechanism was proposed based on the kinetic assay. A directed evolution methodology was designed for glycosyltransferases using Cst-II as the model system. A single mutation, F91Y was found to substantially increase the reaction rate of the enzyme with a fluorescent-coupled acceptor, bodipy-lactose. The crystal structure of this Cst-II F91Y mutant was solved and it revealed an unexpected flip of the tyrosine side chain of Y91 from the core of the enzyme into the solvent region, exposing a hydrophobic pocket which seems to be capable of accommodating the bodipy ring structure. Together with kinetic analyses, the crystallographic study was able to explain the observed increase in the catalytic rate for this novel sugar acceptor. A monofunctional variant of Cst, Cst-I, also isolated from Campylobacter jejuni, was characterized crystallographically and kinetically. The conservation of active site residues supports the proposed mechanism for GT-42 sialyltransferases. The complex structure of the Cst-I enzyme with the donor analogue CMP-3FNeu5Ac provides a platform for molecular modeling of various acceptors into the active sites of Cst-I and Cst-II. The modeling shed lights upon the understanding of differences in substrate specificity. The structures of these complexes will be used as templates to design therapeutic inhibitors against this common human pathogen.
Medicine, Faculty of
Biochemistry and Molecular Biology, Department of
Graduate

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4

Stenbäck, Anders. "Studies of experimental bacterial translocation /". Uppsala : Acta Universitatis Upsaliensis : Univ.-bibl. [distributör], 2005. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-5933.

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5

Cordes, Frank Stephan. "Biophysical studies of bacterial pathogenesis". Thesis, University of Oxford, 2004. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.404113.

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6

Briggs, David Christopher. "Structural studies of bacterial toxins". Thesis, Birkbeck (University of London), 2004. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.414899.

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7

Wilkins, G. M. "Studies on bacterial gene transposition". Thesis, University of Warwick, 1987. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.383407.

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8

Lynch, Anthony Simon. "Studies of bacterial DNA topoisomerases". Thesis, University of York, 1989. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.329651.

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9

Di, Ilio C. "Studies on bacterial glutathione transferase". Thesis, Cranfield University, 1992. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.333472.

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10

Hidese, Ryota. "Studies on Bacterial Dihydropyrimidine Dehydrogenases". Kyoto University, 2011. http://hdl.handle.net/2433/142342.

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Kyoto University (京都大学)
0048
新制・課程博士
博士(農学)
甲第16144号
農博第1880号
新制||農||991(附属図書館)
学位論文||H23||N4614(農学部図書室)
28723
京都大学大学院農学研究科応用生命科学専攻
(主査)准教授栗原達夫, 教授小川順, 教授阪井康能
学位規則第4条第1項該当

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11

Holbourn, Kenneth Paul. "Structural studies on bacterial toxins". Thesis, University of Bath, 2006. https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.436881.

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12

Bergström, Niklas. "Structural studies of bacterial carbohydrate antigens with focus on oral commensal bacteria /". Stockholm : Karolinska institutets bibl, 2002. http://diss.kib.ki.se/2002/91-7349-236-1.

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13

Château, Maarten de. "Functional, structural and evolutionary studies on a family of bacterial surface proteins". Lund : Dept. of Cell and Molecular Biology, Lund University, 1996. http://catalog.hathitrust.org/api/volumes/oclc/38947242.html.

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14

Rozell, Björn. "Immunohistochemical studies of the thioredoxin system". Göteborg : Dept. of Histology, University of Göteborg, 1987. http://catalog.hathitrust.org/api/volumes/oclc/17242526.html.

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15

Sarasanandarajah, Sivananthan. "Multiwavelength fluorescence studies of Bacillus bacterial spores". Thesis, University of Canterbury. Physics and Astronomy, 2007. http://hdl.handle.net/10092/3544.

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Fluorescence techniques are being considered for the detection and identification of bacterial spores. This thesis sets out to empirically characterize the detailed autofluorescence spectroscopic properties of spores and their target molecules. The multiwavelength fluorescence studies from a unique endogenous biomarker, dipicolinic acid (DPA) and its calcium salt (CaDPA) in bacterial spores are found to be useful for fluorescence characterization of spores. A systematic determination of the fluorescence profile of the major chemical components of Bacillus spores and the effect of UV irradiation on them has been performed in dry samples, wet paste and in aqueous solution. The thesis applies reliable tools for accurately describing complex nature of spectral profile from bacterial spores, and for interpreting and identifying their spectral properties. We show that multiwavelength fluorescence technique combined with Principal Component Analysis (PCA) clearly indicates identifiable grouping among dry and wet Bacillus spore species. Differences are also observed between dried, wet and redried spores, indicating the stark effect of hydration on fluorescence fingerprints. The study revealed that changes in fluorescence of spores due to hydration/drying were reversible and supports a recent model of a dynamic and dormant spore structure. The spectra were analysed with PCA, revealing several spectroscopically characteristic features enabling spore species separation. The identified spectral features could be attributed to specific spore chemical components by comparing the spore sample signals with spectra obtained from the target molecules. PCA indicated underlying spectral patterns strongly related to species and the derived components were correlated with the chemical composition of the spore samples. More importantly, we examined and compared the fluorescence of normal spores with a mutant of the same strain whose spores lack DPA. We discovered that the dramatic fluorescence enhancement of Bacillus spores can be caused by UV irradiation in the spectral region of this unique biomarker without any pre treatment. Differences between spectra of spores, spore strains and other biological samples are very marked and are due to the dominance of the dipicolinate features in the spore spectra. This could lead to a cheap, more sensitive, faster and reagentless bacterial spore detector.

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16

Lawrence, Christopher C. "Studies on bacterial proline 4-hydroxylase". Thesis, University of Oxford, 1993. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.358610.

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17

Agnew, Christopher R. J. "Structural studies of bacterial infection proteins". Thesis, University of Bristol, 2011. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.544420.

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18

Hill, C. P. "Structural studies on a bacterial ribonuclease". Thesis, University of York, 1986. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.375429.

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19

Attarataya, Jakrada. "Structural studies of bacterial oxidoreductase enzymes". Thesis, University of Bristol, 2014. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.658865.

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Salmonella are a leading cause of food borne illnesses in humans. Even though symptoms are often mild, these enterobacteria are responsible for over 200,000 deaths worldwide every year. There is considerable concern over the emergence of drug-resistant strains of Salmonella. Lactate dehydrogenase (LDH) from Salmonella typhi is believed to be an essential enzyme for the bacteria and hence a potential drug target. This enzyme is a D-speCific LDH and hence belongs to a class of enzymes much less studied than the more common L-specific LDHs. In this study the expression, structural and functional characterisation of the Salmonella D-LDH enzyme is reported. Crystal structures of the enzyme indicate a number ' of unusual features within its active site that may facilitate specific targeting by novel inhibitors . . Preliminary enzymatic activity data confirm the expected Dspecificity of this enzyme, and that oxamic acid is a weak inhibitor with a Ki of 10.5 mM. The crystal 'structures and enzymatic analyses provide a basis for structure-based design of novel inhibitors. This thesis also describes preliminary studies aimed at determining the structure of a cytochrome P450 enzyme from Jeotga/icoccussp. (oleTJE). This enzyme, a member of the CYP152 family, has peroxygenase activity and can be used to produce alkenes (olefins). It is therefore of interest for the development novel biofuels. Its crystallization is reported, although initial attempts to determine its crystal structure have so far been unsuccessful.

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20

Silva, Avalos Juan G. (Juan Guillermo). "Isolation, Characterization and Physiological Studies of Cyanide-Utilizing Bacteria". Thesis, University of North Texas, 1991. https://digital.library.unt.edu/ark:/67531/metadc278291/.

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Ten bacteria capable of growth on the metal-cyano complex, tetracyanonickelate (II) {K2 [Ni(CN)J } (TCN), supplied as the sole nitrogen source, were isolated. Seven isolates were identified as pseudomonads while the remaining three were classified as Klebsiella species. In addition to TCN, all isolates were able to utilize KCN although it was significantly more toxic. The degradation of TCN was most complete when supplied at growth-limiting concentrations, did not occur when ammonia was present, and resulted in the formation of nickel cyanide [Ni(CN)2] as a degradation product.

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21

Kanso, Sungwan, i n/a. "Molecular Studies of Bacterial Communities in the Great Artesian Basin Aquifers". Griffith University. School of Biomolecular and Biomedical Science, 2004. http://www4.gu.edu.au:8080/adt-root/public/adt-QGU20040219.140509.

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16S rRNA gene analysis has shown that bacterial diversity in the GAB bores studied was limited to the genera Hydrogenobacter in the phylum Aquificae, Thermus in the phylum Deinococcus-Thermus, Desulfotomaculum in the phylum Firmicutes, the alpha-, beta- and gamma-classes of the phylum Proteobacteria and the phylum Nitrospirae. There was no clone closely related to members of the delta-proteobacteria and epsilon-proteobacteria classes detected. The number of bacterial strains directly isolated from the Fairlea and the Cooinda bores were far less than the numbers of distinctive phylotypes detected by the 16S rRNA gene characterisation. In addition none of the bacterial strains directly isolated from the water samples were represented in the 16S rRNA gene clone libraries. Similar discrepancies between the bacterial populations obtained from the 16S rRNA gene analysis and those obtained from direct isolation have been reported in the literature (Dunbar et al., 1999; Kampfer et al., 1996; Suzuki et al., 1997; Ward et al., 1998; Ward et al., 1997). However, in general, the phyla with which the isolates were affiliated were the same as those phyla to which the clones belonged. The environmental changes introduced (by bringing the artesian water up to the surface and exposing it to four types of metal coupons made of carbon steels identified by codes ASTM-A53B, ASTM-A53, AS-1074 and AS-1396 and commonly used in bore casings) led to changes in the bacterial community structures. In general, the species which proliferated in the communities before and after the changes were different. The diversity of the bacterial species in the community decreased following the environmental changes. Clones dominating the clone libraries constructed from newly established bacterial communities also differed from the clones dominating the libraries constructed from the bacterial communities which had existed naturally in the bores. These trends toward change in the bacterial communities were observed at both the Fairlea and the Cooinda bore sites. All four metal types incubated in the Fairlea bore water lost between 3.4 and 4.7% of their original weight. In contrast none of the metals incubated in Cooinda bore water lost weight. Clone library A1 showed that the natural population of the Fairlea bore was dominated by clone A1-3, which represented a novel species related to the isolate boom-7m-04. But after metal incubation (and recording of the metal weight loss), the bacterial community was dominated by clone PKA34B, which has a 95% similarity in its 16S rRNA gene sequence with Desulfotomaculum putei. Desulfotomaculum species are known to cause metal corrosion due to their byproduct H2S. But the low level of phylogenetic relatedness found does not provide enough information to speculate on whether the species represented by clone PKA34B is a member of the genus Desulfotomaculum or not. However, the fact that clone PKA34B dominated the PKA clone library by 50% makes the species it represents a suspected candidate likely to be involved with the metal weight loss at the Fairlea bore. In contrast, clone library 4381 showed that the natural population of the Cooinda bore was dominated by clone 4381-15 representing a species distantly related to a hydrogen oxidiser Hydrogenophaga flava (95% similarity). The dominating clone of the new community formed after metal incubation was clone COO25, which has 99% similarity with Thermus species that have not been reported to be involved with metal corrosion to my knowledge. In this project detection, identification and comparative quantification by 16S rRNA gene-targeted PCR probing with probes 23B and 34B were successfully developed for a Leptothrix-like species and for a Desulfotomaculum-like species represented by clones PKA23B and PKA34B respectively. This method of probing permits a fast, sensitive and reproducible detection, identification and at least a comparative quantification of the bacteria in the environment without the need for culturing. Therefore it is extremely suitable for use in bacterial population monitoring. PCR probing with the 34B probe has a potential commercial use as a means of screening for bores with a potential high risk of corrosion due to this Desulfotomaculum-like species. Direct isolation of bacteria from the GAB water has resulted in the isolation of seven strains from the Fairlea bore and eight from the Cooinda bore. Among these isolates, three novel strains were studied in detail. Reports on the characterisation of strain FaiI4T (T=Type strain) from the Fairlea bore (Kanso & Patel, 2003) and strain CooI3BT from the Cooinda bore have been published (Kanso et al., 2002). The data generated during this project add to our current information and extend our knowledge about the bacterial communities of the GAB's sub-surface environment. This information will provide a basis for further ecological studies of the GAB. Studies on involvement of certain groups of bacteria with the corrosion of metals used in bore casings could provide a foundation for further studies to develop maintenance and managing strategies for the GAB bores.

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22

Kanso, Sungwan. "Molecular Studies of Bacterial Communities in the Great Artesian Basin Aquifers". Thesis, Griffith University, 2004. http://hdl.handle.net/10072/366613.

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16S rRNA gene analysis has shown that bacterial diversity in the GAB bores studied was limited to the genera Hydrogenobacter in the phylum Aquificae, Thermus in the phylum Deinococcus-Thermus, Desulfotomaculum in the phylum Firmicutes, the alpha-, beta- and gamma-classes of the phylum Proteobacteria and the phylum Nitrospirae. There was no clone closely related to members of the delta-proteobacteria and epsilon-proteobacteria classes detected. The number of bacterial strains directly isolated from the Fairlea and the Cooinda bores were far less than the numbers of distinctive phylotypes detected by the 16S rRNA gene characterisation. In addition none of the bacterial strains directly isolated from the water samples were represented in the 16S rRNA gene clone libraries. Similar discrepancies between the bacterial populations obtained from the 16S rRNA gene analysis and those obtained from direct isolation have been reported in the literature (Dunbar et al., 1999; Kampfer et al., 1996; Suzuki et al., 1997; Ward et al., 1998; Ward et al., 1997). However, in general, the phyla with which the isolates were affiliated were the same as those phyla to which the clones belonged. The environmental changes introduced (by bringing the artesian water up to the surface and exposing it to four types of metal coupons made of carbon steels identified by codes ASTM-A53B, ASTM-A53, AS-1074 and AS-1396 and commonly used in bore casings) led to changes in the bacterial community structures. In general, the species which proliferated in the communities before and after the changes were different. The diversity of the bacterial species in the community decreased following the environmental changes. Clones dominating the clone libraries constructed from newly established bacterial communities also differed from the clones dominating the libraries constructed from the bacterial communities which had existed naturally in the bores. These trends toward change in the bacterial communities were observed at both the Fairlea and the Cooinda bore sites. All four metal types incubated in the Fairlea bore water lost between 3.4 and 4.7% of their original weight. In contrast none of the metals incubated in Cooinda bore water lost weight. Clone library A1 showed that the natural population of the Fairlea bore was dominated by clone A1-3, which represented a novel species related to the isolate boom-7m-04. But after metal incubation (and recording of the metal weight loss), the bacterial community was dominated by clone PKA34B, which has a 95% similarity in its 16S rRNA gene sequence with Desulfotomaculum putei. Desulfotomaculum species are known to cause metal corrosion due to their byproduct H2S. But the low level of phylogenetic relatedness found does not provide enough information to speculate on whether the species represented by clone PKA34B is a member of the genus Desulfotomaculum or not. However, the fact that clone PKA34B dominated the PKA clone library by 50% makes the species it represents a suspected candidate likely to be involved with the metal weight loss at the Fairlea bore. In contrast, clone library 4381 showed that the natural population of the Cooinda bore was dominated by clone 4381-15 representing a species distantly related to a hydrogen oxidiser Hydrogenophaga flava (95% similarity). The dominating clone of the new community formed after metal incubation was clone COO25, which has 99% similarity with Thermus species that have not been reported to be involved with metal corrosion to my knowledge. In this project detection, identification and comparative quantification by 16S rRNA gene-targeted PCR probing with probes 23B and 34B were successfully developed for a Leptothrix-like species and for a Desulfotomaculum-like species represented by clones PKA23B and PKA34B respectively. This method of probing permits a fast, sensitive and reproducible detection, identification and at least a comparative quantification of the bacteria in the environment without the need for culturing. Therefore it is extremely suitable for use in bacterial population monitoring. PCR probing with the 34B probe has a potential commercial use as a means of screening for bores with a potential high risk of corrosion due to this Desulfotomaculum-like species. Direct isolation of bacteria from the GAB water has resulted in the isolation of seven strains from the Fairlea bore and eight from the Cooinda bore. Among these isolates, three novel strains were studied in detail. Reports on the characterisation of strain FaiI4T (T=Type strain) from the Fairlea bore (Kanso & Patel, 2003) and strain CooI3BT from the Cooinda bore have been published (Kanso et al., 2002). The data generated during this project add to our current information and extend our knowledge about the bacterial communities of the GAB's sub-surface environment. This information will provide a basis for further ecological studies of the GAB. Studies on involvement of certain groups of bacteria with the corrosion of metals used in bore casings could provide a foundation for further studies to develop maintenance and managing strategies for the GAB bores.
Thesis (PhD Doctorate)
Doctor of Philosophy (PhD)
School of Biomolecular and Biomedical Sciences
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23

Zeng, Muling. "Bacterial cellulose: fabrication, characterization and biocompatibility studies". Doctoral thesis, Universitat Autònoma de Barcelona, 2014. http://hdl.handle.net/10803/284146.

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En marzo de 2011, apliqué a una beca del CSC (Consejo de Becas de China), en cooperación con la Universitat Autònoma de Barcelona (UAB). Después de medio año, conseguí la beca y comencé mi tesis doctoral bajo la supervisión de la Dra. Anna Roig y la Dra. Anna Laromaine. Mi proyecto asignada era en celulosa bacteriana: su síntesis, caracterización y estudios de biocompatibilidad. La celulosa bacteriana es un polisacárido de fuentes renovables, y puede ser producida por algunos tipos de bacterias en la naturaleza. Presenta propiedades químicas y físicas notables, incluyendo una alta pureza química y cristalinidad, una red de nanofibras, porosa, alta capacidad de absorción de agua y resistencia mecánica. La celulosa bacteriana se utiliza para una amplia variedad de aplicaciones comerciales. Por otra parte, la celulosa bacteriana es biocompatible con afinidad biológica y biodegradabilidad, que suscita la atención de investigadores en el área de la biomedicina. El primer objetivo de mi tesis fue aprender a producir películas de celulosa bacteriana y encontrar estrategias para controlar sus propiedades. Un segundo objetivo fue el desarrollo de métodos para la fabricación de nanocompuestos basedos con celulosa bacteriana. El objetivo final estudia la biocompatibilidad de las capas de celulosa bacteriana que hemos producido y su utilidad como soportes tridimensionales para el crecimiento celular. Así se pretende establecer una plataforma para el estudio de la interacción de células y nanopartículas en un entorno 3D más realista. Durante el primer año, se realizó la puesta a punto en el laboratorio del sistema para producir capas de celulosa bacteriana a partir de dos cepas bacterianas y secarlas a partir de tres métodos de secado:a temperatura ambiente, secado por liofilización y secado supercrítico. Por otra parte, se realizó la caracterización completa de las capas de celulosa bacteriana: su porosidad, la transparencia, la capacidad de absorción de agua y las propiedades mecánicas que se podían controlar seleccionando la cepa bacteriana y el método de secado. En el segundo año, se sintetizó la celulosa bacteriana compuesta con nanopartículas por el método asistida por microondas como materiales de celulosa funcionales novedosos. Este método es eficiente y rápido, forma un recubrimiento de las capas de celulosa bacteriana por nanopartículas de forma homogénea y controlable. Secando las capas de celulosa utilizando diferentes rutas, se puede controlar la cantidad final del contenido de las nanopartículas en los materiales compuestos. Así capas con dominios hidrófobos / hidrófilos favorecían el anclaje de nanopartículas de forma selectiva para crear materiales de celulosa más complejos y funcionales. En este último año, se ha llevado a cabo el estudio de la biocompatibilidad de las capas de celulosa bacteriana in vitro. Aunque la celulosa bacteriana se considera generalmente un material no citotóxico, su biocompatibilidad es un requisito importante para su uso en aplicaciones biológicas y médicas y no ha sido evaluado completamente. Además se fabricó una estructura de celulosa bacteriana 3D mejorada. La tesis se estructura en seis capítulos. Capítulo 1 proporciona una introducción a la celulosa bacteriana. Capítulo 2 ofrece una descripción detallada de la fabricación de capas de celulosa bacteriana (BCF). Capítulo 3 se centra en la síntesis de compuestos de celulosa bacteriana funcionales que incorporan nanopartículas. Capítulo 4 presenta estudios de biocompatibilidad de la celulosa bacteriana como estructura 2D y 3D para estudios celulares in vitro. Capítulo 5 se enumeran las principales conclusiones derivadas de la presente tesis y algunas sugerencias para el trabajo futuro.Capítulo 6 recoge información sobre el autor y las publicaciones durante el doctorado.
In March 2011, I started the application of a scholarship from CSC (Chinese Scholarship Council), which cooperated with the Universitat Autònoma de Barcelona (UAB). After about half year, I secured the scholarship and began my doctoral thesis under the supervision of Dr. Anna Roig and Dr. Anna Laromaine. My project assignment was on bacterial cellulose: fabrication, characterization and biocompatibility studies. Bacterial cellulose is a renewable polysaccharide, which is produced by some types of bacteria in nature. It presents remarkable chemical and physical properties, including high chemical purity and crystallinity, nano-scale fibre network, porosity, high water absorption capacity and mechanical strength. Bacterial cellulose is being used for a wide variety of commercial applications, for example textiles, cosmetics, food products and other technical areas. Furthermore, bacterial cellulose is also biocompatible with excellent biological affinity and biodegradability, which is drawing immense attention from the bio and medical area researchers. The objective of my thesis was to learn how to produce bacterial cellulose films and find strategies to control their properties. A second objective was to developed methods to fabricate nanocomposites based on bacterial cellulose. The final objective was related to prove the biocompatibility of the in-house produced bacterial cellulose films and to be able to use them as three-dimensional scaffolds for cell in-growth. In this way setting up a platform that will allow us to study the interaction of cells and nanoparticles in a realistic 3D environment. During the first year, a lab set-up was successfully built to produce bacterial cellulose from two bacterial strains and three methods of drying were accessed to dry the thin films; at room temperature, freeze drying and supercritical drying. Moreover, the full characterization of bacterial cellulose films was accomplished: their porosity, transparency, water absorption capacity and mechanical properties were tuned by selecting the bacterial strain and the drying method. In the second year, bacterial cellulose composited with nanoparticles as novel functional cellulose materials were synthesized by microwave-assisted method. This method is efficient and fast to form a homogenous conformal and controllable coating of nanoparticles on the bacterial cellulose films. By drying the cellulose films using different routes, the final amount of the nanoparticles content in the composites can be controlled. Furthermore, those films were patterned with hydrophobic/hydrophilic domains and selectively anchored nanoparticles to create more complex and functional cellulose composites. During the last year, an investigation of the biocompatibility of the bacterial cellulose films in vitro was performed. Although bacterial cellulose is generally considered non-cytotoxic material, its biocompatibility as a major requirement for the use in biological and medical applications has not been fully evaluated. Furthermore, an improved 3D bacterial cellulose scaffold was fabricated. The thesis is organized into six chapters. Chapter 1 provides an introduction to bacterial cellulose. Chapter 2 describes a detailed description of the fabrication of bacterial cellulose films (BCFs). Chapter 3 focuses on the synthesis of functional bacterial cellulose composites incorporating nanoparticles. Chapter 4 presents the studies of bacterial cellulose biocompatibility as 2D and 3D scaffold for cell studies in vitro. Chapter 5 lists the main conclusions derived from the present thesis and some suggestions for the future work. Chapter 6 gathers information about the author and the publications during the Ph.D. studies.

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Noursadeghi, Mahdad. "Studies of innate immunity to bacterial infection". Thesis, Imperial College London, 2003. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.406424.

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Bond, Peter J. "Simulation studies of bacterial outer membrane proteins". Thesis, University of Oxford, 2005. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.419258.

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Onakunle, Ojo Adegboyega. "Studies on the enantioselectivity of bacterial lactonases". Thesis, University of Kent, 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.362189.

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Gibson, Kevin J. C. "Structural and biochemical studies on bacterial lipoglycans". Thesis, University of Newcastle Upon Tyne, 2003. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.397343.

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Conroy, Matthew James. "NMR studies of bacterial light-harvesting complexes". Thesis, University of Sheffield, 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.298888.

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Trambaiolo, Daniel Marco. "Crystallographic studies of bacterial cell division proteins". Thesis, University of Cambridge, 2008. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.611977.

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Lloyd, Diarmuid Padraig. "Microscopic studies of surface growing bacterial populations". Thesis, University of Edinburgh, 2015. http://hdl.handle.net/1842/10509.

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In this thesis, I present three microscopy studies of surface growing Escherichia coli (E. coli ) microcolonies. All experiments were carried out by growing microcolonies on agarose pads, and imaging their growth using phase contrast, fluorescence and confocal microscopy. In the first project, the importance of spatial structure and growth strategies between competing populations of E. coli was studied. An agarose pad was seeded with bacterial cells and their colonisation success tracked. Cell lag-times and local cell density were found to play important roles in determining the eventual success of a colony. Arrangements of neighbouring cells were found to be partially responsible at high cell densities. These results were reproduced using a simple simulation, which also highlighted the importance of exponential expansion in determining colonisation success. The second project investigates the effect of confinement on growing microcolonies restricted to one plane (2d growth). Colonies were grown in agarose microchannels with different aspect ratios, and in unconfined environments. In particular internal physical colony structure and genealogical structure was studied by using single-cell tracking. Results showed that relatedness between cells was directionally biased (cells tended to be more closely related to cells at their poles, than to their side) regardless of the amount of spatial restriction. Furthermore, confinement caused cells to align with each other more, and induced high cell velocities at the colony edges driven by cell expansion. In the final project, growth of secondary layers in growing colonies of E. coli was studied. Cells initially grew as a monolayer, before invading the agarose bulk, producing a secondary layer. By analysing time-lapse movies, this layer was found to initially expand rapidly well in excess of cell growth rates and initial colony expansion rates, before slowing down. The initial secondary growth rate likely depends on the colony area at agarose invasion. Furthermore, the colony area when colonies invaded the agarose depended on their rate of growth, suggesting a complex interplay between forces exerted by the agarose, and by the colony.

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31

Kawasaki, Haruhiko. "STUDIES ON PLASMIDS DETERMINING BACTERIAL HALOACETATE DEHALOGENASES". Kyoto University, 1985. http://hdl.handle.net/2433/78183.

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Ramesar, Rajkumar Sewcharan. "Developmental genetic studies on Thiobacillus ferrooxidans". Doctoral thesis, University of Cape Town, 1988. http://hdl.handle.net/11427/26235.

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Thiobacillus ferrooxidans is an industrially important bacterium which is used in bioleaching operations. The work reported in this investigation extends current knowledge of the genetics of this organism. Conjugation was attempted as a means for plasmid DNA transfer to T. ferrooxidans. Recombinant T. ferrooxidans plasmids, pDER401 and pDER405, were shown to code for mobilization and replication functions in Escherichia coli and Thiobacillus novellus strains. The plasmids were mobilizable at high frequency by the IncP plasmid, R68.45. Attempts to transfer the T. ferrooxidans recombinant plasmids directly from E. coli to T. ferrooxidans were unsuccessful. In multistage mating experiments, plasmid DNA was transferred from E. coli to T. novellus, and from T. novellus to Thiobacillus intermedius. However, in subsequent matings, plasmid transfer from these thiobacilli to T. ferrooxidans could not be shown. A genomic library of T. ferrooxidans ATCC 33020 was constructed in the plasmid vector, pEcoR251, for the purpose of cloning a recA-like gene from this organism. The library consisted of approximately 1,78 X 10⁴ clones carrying chromosomal DNA fragments of about 3-12 kilobases (kb). The library was successfully screened for functional complementation of E. coli auxotrophic mutants. Clones that conferred resistance to methyl methane sulfonate (MMS), a DNA-damaging agent, were isolated in an E. coli recA⁻ mutant. In an attempt to clone a homologous marker, T. ferrooxidans ATCC 33020 was mutated to rifampicin resistance (Rifʳ) and DNA from the mutant strain was used in the construction of plasmid- and cosmid-based libraries. The plasmid library contained approximately 1,35 X 10⁴ clones with inserts of about 1-13 kb. The cosmid library consisted of approximately 8.2 X 10³ colonies, 4.0 X 10⁴ in vitro packaged cosmids, and an amplified in vivo-packaged cosmid lysate containing approximately 1.82 X 10¹¹ infectious particles, carrying inserts of about 35-55 kb. Complementation of E. coli auxotrophic mutants was observed with the plasmid and cosmid library of the T. ferrooxidans Rifʳ strain. Screening both libraries for a Rifʳ marker was unsuccessful. Three recombinant plasmids, pRSR100, pRSR101, and pRSR102, each containing the functional analogue of the E. coli recA gene, were isolated from the plasmid-based genomic library of T. ferrooxidans ATCC 33020. The plasmid, pRSR100, was used for further characterization of the cloned recA-like gene. pRSR100 complemented defects in DNA repair and homologous recombination in an E. coli recA- strain. Antiserum raised against E. coli RecA⁻ protein reacted with two protein bands with an apparent Mᵣ of approximately 40 000 and 38 000 in extracts of the recA deletion mutant, E. coli JK696, containing pRSR100. A single band with an apparent Mᵣ of approximately 40 000 was detected in T. ferrooxidans cell extracts with the E. coli RecA antiserum. The nucleotide sequence of the T. ferrooxidans recA gene has been determined. No SOS box characteristic of LexA- regulated promoters could be identified in the 196-bp region upstream of the coding region. The T. ferrooxidans recA gene specifies a protein of 346 amino acids that has 66% and 69% homology to the RecA proteins of E. coli and P. aeruginosa, respectively. Most amino acids that have been identified as being of functional importance in the E. coli RecA protein are conserved in the T. ferrooxidans RecA protein. Although some amino acids that have been associated with ATPase and constitutive protease activity have been substituted, the cloned protein has retained these activities. The cloned recA gene was expressed in E. coli from both the λ Pᵣ and lac promoters. However, no expression from the 2.2 kb T. ferrooxidans DNA preceding the gene was evident.

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33

Prayitno, Slamet Budi. "Studies of bacteria causing prawn disease in Indonesia with special emphasis on luminous bacterial disease". Thesis, Bangor University, 1994. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.483433.

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McClellan, J. A. "Studies on Escherichia coli-mutabile". Thesis, University of Leeds, 1985. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.355195.

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Choudhury, Hassanul Ghani. "Structural and functional studies of bacterial detoxification mechanisms". Thesis, Imperial College London, 2013. http://hdl.handle.net/10044/1/39387.

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Bacterial multidrug resistance is a prominent issue with an ever-increasing amount of drugs becoming ineffective in treating bacterial infections. The elucidation of the 3D structure of bacterial resistance proteins has become extremely important in the development of new antimicrobials. Bacteria can sense external stimuli and regulate the expression of genes that can confer antibiotic resistance. Response regulators are capable of binding DNA both in their phosphorylated and unphosphorylated states, upregulating gene expression. The crystal structure of the unphosphorylated dimeric Escherichia coli BaeR in an intermediate state has been determined. One subunit of the dimer is in an 'active-like' conformation and the other in an inactive conformation. The dimer is formed by a domain swap, which is supported by our small angle X-ray scattering data. This structure provides a possible mechanism on how an unphosphorylated response regulator is capable of binding DNA. Bacteria can detoxify toxic chalcogens, selenium and tellurium, by methylation. The structure of the Escherichia coli TehB has been determined. From our functional analysis, TehB is capable of methylating different chalcogen oxyanions, making it a powerful detoxifying protein. Based on the structure, mutagenesis and kinetic data, we proposed a reaction mechanism for chalcogen detoxification. This data provides the first molecular understanding of the detoxification process of chalcogens by bacteria. In order to survive, certain bacteria under nutrient starved conditions produce antimicrobial peptides (e.g. bacteriocins and microcins) that target closely related species. They have dedicated ABC transporters to export the toxins out of the cells producing them. The crystal structure of the microcin J25 ABC exporter McjD has been determined. Our biochemical data show McjD has a high specificity towards its substrate. In addition, McjD adopts a new 'nucleotide-bound outward-occluded' conformation. Comparison with other ABC exporters has allowed us to propose an intermediate step in ABC exporter mechanism.

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36

Diab, Asim Eltayeb. "Experimental bacterial meningitis : studies on immunopathogenesis and immunoregulation /". Stockholm, 1998. http://diss.kib.ki.se/1998/91-628-3008-2.

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Tanabe, Mikio. "Structural, functional and immunological studies of bacterial transporters". Thesis, Imperial College London, 2006. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.428677.

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38

Qua, A. R. "Anti-bacterial structure-function relationship studies on melittin". Thesis, Queen's University Belfast, 2005. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.419557.

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Cheesman, Myles Richard. "Spectroscopic studies of nickel ions in bacterial proteins". Thesis, University of East Anglia, 1988. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.327824.

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40

Cox, Charles. "Structural and functional studies of bacterial mechanosensitive channels". Thesis, Cardiff University, 2012. http://orca.cf.ac.uk/44553/.

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Bacterial mechanosensitive (MS) channels represent the most primitive form of MS channels gating solely in response to changes in bilayer tension. The two main families of bacterial MS channels are MscS and MscL. In concert these channels are imperative to obviate the effects of toxic downshocks in external osmolarity. While osmoprotection is the best characterised physiological role for these channels the genetic diversity of this family hints at as yet undiscovered physiological functions. This thesis explores one such possibility, showing that MscS expression provides a selective advantage in the presence of cell wall attack with knockout E. coli strains of MscS having increased susceptibility to cell wall targeting antibiotics. This thesis also shows, using the Ca2+ sensitive photoprotein aequorin, that on gating these channels become not only a gateway for solute efflux but also a conduit for ion entry. In particular, influx of Ca2+ may represent a physiologically relevant signal but more importantly this finding may pave the way for a high-throughput screen for novel MS channel activators which would be of potential value as lead compounds for antibiotics. Furthermore, this thesis demonstrates that in the presence of divalentcations (Ca2+ & Ba2+) MscS exhibits increased anion selectivity, rectification and eight distinct long-lived subconducting states at hyperpolarising membrane potentials. In an attempt to identify the structural basis of these subconducting states the first single residue MscS mutants that display altered anion selectivity are reported. This selectivity, in contrast to voltage-gated K+, Na+ and Ca2+ channels, is not determined by residues in the pore region but rather by charged residues in the cytoplasmic domain and is likely conserved throughout the MscS family.

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41

Duman, Ramona Elena. "Structural studies of bacterial Lon ATP-dependent proteases". Thesis, University of Cambridge, 2010. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.609086.

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Back, Catherine R. "Structural and functional studies of oral bacterial adhesins". Thesis, University of Bristol, 2014. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.650102.

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Biofilm and complex microbial community formation are fundamental processes in colonisation of the human oral cavity. Defining the mechanisms by which microbes interact with each other will lead to better understanding of how these communities are assembled. Many species of oral bacteria express cell-surface proteins (adhesins) that promote colonisation and infection through host receptor recognition. Streptococcus gordonii is ' a commensal primary coloniser of the oral cavity, and is also implicated as a leading cause of infective endocarditis . S. gordonii expresses a number of cell-surface proteins that enable the bacteria to colonise a variety of sites throughout the human body and to coaggregate with other microorganisms in biofilm community development. These include two Antigen I/ ll-family proteins, 'termed SspA and SspB, that have been implicated in colonisation and pathogenic mechanisms utilised by S. gordonii. The first aim of this study was to determine the molecular basis of interaction between two early colonisers of the oral cavity, S. gordonii and Actinomyces oris. This work identified a novel primary coloniser interaction involving recognition by SspB of cell surface polysaccharide expressed by A. oris. Another adhesin expressed by S. gordonii is CshA, which forms fibrils that extend away from the bacterial cell surface, interacts with fibronectin, and mediates coaggregation with other oral microorganisms in the formation of biofilm communities. However, the molecular mechanisms that underpin these interactions were unknown. The second aim of this study was to obtain detailed structural information about CshA, and to define functional regions relevant to S. gordonii colonisation and pathogenesis. This work showed that CshA had a unique structure with new protein folds, not previously reported for bacterial adhesins, and identified specific functional regions. Better understanding of molecular mechanisms involved in bacterial colonisation will assist development of new clinical interventions for diseases caused by oral microorganisms.

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43

Walford, Gillian. "NMR and conformational studies of bacterial polysaccharide antigens". Thesis, University of Cambridge, 1993. https://www.repository.cam.ac.uk/handle/1810/272377.

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Costa, Jill Bonham. "Structural studies of some viscous, acidic bacterial exopolysaccharides /". The Ohio State University, 1991. http://rave.ohiolink.edu/etdc/view?acc_num=osu1487687959968165.

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Breuleux, Claire Emilie. "Studies on bacterial lung infections in cystic fibrosis". Thesis, Aston University, 2000. http://publications.aston.ac.uk/12359/.

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The major cause of death in CF is a continuous inflammation of the lungs colonised with Pseudomonas aeruginosa and occasionally also with Burkholderia cepacia. A combination of serum IgG to LPS and serum PCT levels were found to be good markers for detection of early colonisation with P. aeruginosa. Colomycin sulphomethate (colistin E) is one of the antibiotics used to treat P. aeruginosa infections in CF. Electrophoretic methods were developed to monitor the rate of conversion of colomycin sulphomethate to the active form of the drug. Antimicrobial activity towards P. aeruginosa was generated as the sulphomethate substituents were released. Clinical resistance of P. aeruginosa to colomycin is rare, but a number of isolates have been isolated. Twelve colomycin-resistant clinical isolates were investigated to determine the mechanism of resistance. It was found that the low level of resistance was due to over expression of outer membrane protein H (OprH) in 5 isolates. A novel mechanism of resistance involving modification of the phosphate groups in LPS was identified in one of the isolates. Drugs which reduce inflammation in infected CF lungs would be of great advantage for therapy. Reducing inflammation would preserve the lung function and increase the quality of life for CF patients. Antibiotics like tetracyclines, macrolides and polymyxins were tested for their potential anti-inflammatory effects using cultured human monocytic (U937) cells which secrete the pro-inflammatory cytokines IL1- and TNF- in response to LPS from P. aeruginosa and B. cepacia. It was found that tetracyclines, and especially doxycycline, are good inhibitors of cytokine release by U937 cells and therefore could reduce the inflammatory cascade.

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Malasarn, Davin Sternberg Paul W. "Molecular and environmental studies of bacterial arsenate respiration /". Diss., Pasadena, Calif. : California Institute of Technology, 2007. http://resolver.caltech.edu/CaltechETD:etd-02232007-132917.

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Malik, A. N. "Genetic studies with Pseudomonas syringae pathovar pisi". Thesis, University of Greenwich, 1985. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.354390.

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Hill, Russell. "Gene cloning studies in two nocardioform bacteria". Doctoral thesis, University of Cape Town, 1988. http://hdl.handle.net/11427/21896.

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Bibliography: pages 147-177.
Nocardioforms are Gram-positive, aerobic actinomycetes and are a metabolically diverse group which produce antibiotics, useful enzymes, are important in the biotransformation of organic compounds and the decomposition of organic wastes and are important medically. A gene cloning vector designated pLR591 was constructed from the broad host range, multicopy Streptomyces plasmid pIJ702 and the positive selection Escherichia coli plasmid pEcoR251. This plasmid has useful features for the construction of actinomycete genomic libraries. Cloning of DNA into the unique Bg1II endonuclease site of pLR591 inactivated the lethal EcoRI gene derived from pEcoR251, thereby selecting for recombinant plasmids containing inserted DNA. The thiostrepton resistance gene derived from pIJ702 was shown to be functional in Streptomyces lividans enabling selection of recombinant pLR591 plasmids containing foreign DNA in S. lividans. The vector pLR591 therefore functions as a positive selection Streptomyces-E. coli shuttle vector facilitating construction of actinomycete genomic libraries in E. coli and subsequent transfer of recombinant plasmids into S. lividans.

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Yang, Dong. "Structural studies of terpenoid biosynthesis and bacterial cell division". [College Station, Tex. : Texas A&M University, 2006. http://hdl.handle.net/1969.1/ETD-TAMU-1737.

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Panjikar, Santosh. "Crystallographic studies of bacterial single stranded DNA-binding proteins". [S.l.] : [s.n.], 2001. http://deposit.ddb.de/cgi-bin/dokserv?idn=964154366.

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