No description available.

Diss. (sammanfattning) Umeå : Univ., 2007. / Härtill 6 uppsatser.

Assisting smokers to quit and stay quit is the most important intervention clinicians can undertake to improve the length and quality of life of patients who use tobacco. The chronic, relapsing nature of tobacco dependence complicates tobacco treatment. Tobacco treatment counseling programs provide on-going support to help patients avoid relapse. Assistance with a referral increases the likelihood that patients will participate in counseling, but few clinicians regularly assist with referrals to tobacco treatment programs. This dissertation examined health system processes and clinician attitudes that influence the likelihood that clinicians will refer their patients for tobacco treatment counseling. Three papers examined health system processes, clinician attitudes, and frequency of referrals. A systematic review of the literature was conducted to evaluate strategies to increase the frequency of clinician referrals and effects on quit rates in primary care. The most effective strategies were those that combined clinician education with integrated referral systems. Integrated referral systems include non-clinician staff and clinician and staff prompts with algorithms or protocols for referrals. The second paper reports the findings of a pilot study using qualitative methods to explore experiences and strategies used for tobacco treatment by clinician champions practicing in independent primary care practices. Tobacco champions (N = 11) described experiences counseling patients but not assistance with referrals. Themes identified were: sources of knowledge and experience, understanding dependence, role perception, and treatment strategies. The final paper reports the findings of a cross- sectional, non-experimental study to examine clinician attitudes toward counseling, health system processes that facilitate referrals, and referrals to tobacco treatment counseling. Attitudes about the efficacy of tobacco counseling and the presence of processes that facilitate referrals predicted referrals. Clinicians sampled in these studies held favorable attitudes toward tobacco treatment but lacked confidence in the efficacy of community-based counseling for tobacco treatment. These findings have implications for health care policies to improve integration of processes that facilitate referrals and clinician education that includes information about counseling resources to improve chronic care for the treatment of tobacco dependence.

The spirochete Borrelia hermsii avoids the immune response of its mammalian host through multiphasic antigenic variation. Serotype specificity is determined by Variable major proteins (Vmp), in the outer membrane. Through a non reciprocal recombination between linear plasmids, a formerly silent vmp gene replaces another vmp gene at a telomeric expression locus downstream from a common expression site. B. hermsii before and after the switch from serotype 7 to serotype 21, was examined in detail. The nucleotide sequence of the vmp7 and vmp21 genes and flanking regions was determined. The vmp7 and vmp21 are 77% identical in their coding sequence, and the deduced translation products are 63% identical. No antigenic cross reactivity is observed between Vmp7 and Vmp21. This suggests a folding of the proteins in which the similar regions are buried, and not exposed when it is presented at the bacterial surface. Vmp7 and Vmp21 have consensus sequences of prokaryotic lipoproteins and are processed as such when expressed in E. coli. The 5' regions of silent and expressed vmp7 and vmp21 were compared. Silent and active vmp7 and vmp21 genes shared a block of homologous sequence at their 5' ends. Sequences upstream of silent vmp7 and vmp21 genes lacked a promoter and differed substantially from each other. In this antigenic switch a vmp gene was activated by a recombination event which placed it downstream of a promoter. The vmp gene promoter is preceded by a poly(dT dA) ran and three imperfectlyrepeated elements of 2 kb. Each of the 2 kb repeats contains inverted repeats of approximately 0.2 kb at their termini. There is no evidence of the presence of similar elements elsewhere in the genome of B. hermsii. One or more of these elements may stimulate vmp gene switch or expression. The African relapsing fever species Borrelia crocidurae and the American species B. hermsii display many similarities. In both species the vmp genes are localised to linear plasmids, and the vmp genes are activated on the transcriptional level. The nucleotide sequence of their expression sites, however, are not related. Still, the possibility that the switch is mechanistically similar in B. crocidurae and B. hermsii, cannot be ruled out. The binding of B. crocidurae causes aggregation of erythrocytes around the spirochete. The aggregation is reminiscent of the erythrocyte rosetting seen in malarial infections. The erythrocytes at the B. crocidurae surface may protect them from clearance by the host. Thus, the rosetting may constitute an additional mechanism in B. crocidurae for the evasion of the immune reaction. / <p>Diss. (sammanfattning) Umeå : Umeå universitet, 1994, härtill 5 uppsatser.</p> / digitalisering@umu

Thesis (Ph. D.)--Virginia Commonwealth University, 2007. / Prepared for: Dept. of Microbiology and Immunology. Bibliography: leaves 127-137. Available online via the Internet.

Relapsing fever Borrelia are transmitted through saliva of argasid ticks. Given the 10 to 60 minute long bloodmeal, we characterized the rapidity of Borrelia turicatae transmission from their vector Ornithodoros turicata. Infection rates were compared in mice when cohorts of ticks fed to repletion on animals in which groups of O. turicata were removed 15 seconds after attachment. Infection was evaluated by examining the blood by dark field microscopy, quantitative PCR, and serological responses generated against B. turicatae. Scanning electron microscopy was also performed on cryofractured tick salivary glands to determine spirochete localization. Dissemination of B. turicatae into murine blood was evaluated by removing the bite site after ticks engorged. Our findings indicate that B. turicatae is localized in the lumen of salivary gland acini of O. turicata and transmission to and dissemination in the mammal occur shortly after tick attachment and do not require a complete bloodmeal.

En Afrique, les fièvres récurrentes à Borrelia sont des pathologies négligées transmises par les arthropodes et responsables de septicémie mortelle et d'autres manifestations cliniques, en particulier d'avortement chez les femmes enceintes. Quatre Borrelia différentes sont actuellement cultivées de prélèvements cliniques et de vecteurs, il s'agit de Borrelia crocidurae, Borrelia duttonii, Borrelia recurrentis et Borrelia hispanica. Ces différentes espèces ont été initialement séparées les unes des autres sur la base de leur répartition géographique et de leur vecteur. Au cours de ce travail de thèse, nous avons réalisé le séquençage et l'annotation du génome de Borrelia crocidurae. Ceci nous a permis de comparer le génome de trois espèces séquencées: Borrelia crocidurae, Borrelia duttonii et Borrelia recurrentis. Cette comparaison indique que ces trois espèces sont extrêmement proches comme cela avait déjà été montré par la comparaison de la séquence du gène 16S ARN ribosomal de chacune de ces espèces, montrant une similarité comprise entre 99,7 et 99,9%. L'analyse de données génomiques permet de conforter les résultats antérieurs basés sur l'analyse de quelques gènes et de proposer que ces trois espèces ne forment qu'une seule espèce génomique présentant trois écotypes avec une relative spécificité de vecteur, de répartition géographique et d'évolution clinique. La très grande similitude entre Borrelia crocidurae, Borrelia duttonii et Borrelia recurrentis constitue un obstacle pour la mise au point de techniques de diagnostic direct de ces espèces dans les prélèvements de vecteur ou dans les prélèvements humains. / In Africa, relapsing fever borreliae are neglected arthropod-borne pathogens causing mild to deadly septicemia. Borrelia crocidurae, Borrelia duttonii, Borrelia hispanica and Borrelia recurrentis are the currently cultured causative agents in Africa. The relapsing fever borreliae species were initially distinguished one from another on the basis of geography and vector. we performed the genome sequencing of Borrelia crocidurae. At the genomic level the four species are highly similar as illustrated by the 16S rRNA gene sequence of each species, resulting in an overall high sequence similarity of 99.7 to 99.9% between the four species. Genomic analyses of Borrelia crocidurae, Borrelia duttonii and Borrelia recurrentis further indicated that they in fact forming an unique bacterial species, each one of the three species could be regarded as an ecotype of an unique species with preferential arthropod vector, geographic distribution and clinical outcome, rather than an unique bacterial species. The high similarity between species remained an obstacle for the diagnosis at the species level. Currently the identification of relapsing fever borreliae relies upon a few phenotypic traits and the detection of single nucleotide polymorphisms in the 16S rRNA and flabB, glpQ genes and the 16S-23S ribosomal RNA intergenic spacer (IGS). In this study, based on comparative genomic anaylsis between the published genomic sequence and partial sequence in the genbank, we developed multiplex, quantitative real-time PCR detecting any relapsing fever Borrelia and specifically B. crocidurae, B. hispanica and B. duttonii/B. recurrentis.

Relapsing fever (RF) is caused by different species of Borrelia transmitted by soft ticks or by the human body louse. Illness is characterized by reappearing peaks of high concentrations of spirochetes in blood, concordant with fever peaks separated by asymptomatic periods. Neuroborreliosis is one of the most severe manifestations of RF borreliosis. To understand the immune response during early RF, we analyzed immune cells in brain and kidney of mice infected with B. crocidurae during the acute infection. Our results indicate that brain defense is comprised primarily of innate immune cells. Despite the infiltration of innate immune cells, Borrelia was not completely eradicated. A failure of the host brain to clear the bacteria may give the pathogen a niche where it can persist. Using our mouse model, we revealed that Borrelia duttonii could persist in the mouse brain for up to 270 days, without being present in the circulation. The infection was silent with no change in host gene expression, and the spirochetes could re-enter the circulation after immunosuppression. We propose that the brain is used by the pathogen to evade host immunity and serves as a possible natural reservoir for B. duttonii, a spirochete that has rarely been found in any mammalian host other than man. Borrelia-induced complications during pregnancy have been reported, and are especially common in RF. In our established mouse model of gestational RF, we could show that the fetuses suffered from severe pathology and growth retardation, probably as a consequence of placental destruction. We could also show trans-placental transmission of the bacteria leading to neonatal RF. Surprisingly, pregnant dams had a lower bacterial load and less severe disease, showing that pregnancy has a protective effect during RF. We have used the gestational RF model to investigate host factors favoring disease resolution. Because the spleen is the primary organ responsible for trapping and removing blood-borne pathogens, we have compared temporal changes in spleen immune cell populations and cytokine/chemokine induction during the infection. Spleens of pregnant mice had earlier neutrophil infiltration, as well as faster and higher production of pro-inflammatory mediators. This rapid, robust response suggests a more effective host defense. Thus, an enhanced pro-inflammatory response during pregnancy imparts a distinct advantage in controlling the severity of relapsing fever infection.

Borrelia is a Gram-negative, corkscrew-shaped bacterium transmitted by infected ticks or lice. Borreliae are subdivided into pathogens of two diseases: Lyme disease, caused mainly by B. burgdorferi, B. afzelii and B. garinii; and relapsing fever caused primarily by B. duttonii, B. hermsii, B. recurrentis or B. crocidurae. Both diseases differ in their manifestations, duration times and dissemination patterns. Antibiotics are the major therapeutics, although unfortunately antibiotic treatment is not always beneficial. To date, drug resistance mechanisms in B. burgdorferi are unknown. Transporters of the resistance-nodulation-division (RND) family appear to be involved in drug resistance, especially in Gram-negative bacteria. They consist of three components: a cytoplasmic membrane export system, a membrane fusion protein (MFP), and an outer membrane factor (OMP). The major antibiotic efflux activity of this type in Escherichia coli is mediated by the tripartite multidrug resistance pump AcrAB-TolC. Based on the sequence homology we conclude that the besA (bb0140), besB (bb0141) and besC (bb0142) genes code for a similar efflux system in B. burgdorferi. We created a deletion mutant of besC. The minimal inhibitory concentration (MIC) values of B. burgdorferi carrying an inactive besC gene were 4- to 8-fold lower than in the wild type strain. Animal experiments showed that the besC mutant was unable to infect mice. Black lipid bilayer experiments were carried out to determine the biophysical properties of purified BesC. This study showed the importance of BesC protein for B. burgdorferi pathogenicity and resistance to antibiotics, although its importance in clinical isolates is not known. Due to its small genome, Borrelia is metabolically and biosynthetically deficient, thereby making it highly dependent on nutrients provided by their hosts. The uptake of nutrients by Borrelia is not yet completely understood. We describe the purification and characterization of a 36-kDa protein that functions as a putative dicarboxylate-specific porin in the outer membrane of Borrelia. The protein was designated as DipA, for dicarboxylate-specific porin A. DipA was biophysically characterized using the black lipid bilayer assay. The permeation of KCl through the channel could be partly blocked by titrating the DipA-mediated membrane conductance with increasing concentrations of different organic dicarboxylic anions. The obtained results imply that DipA does not form a general diffusion pore, but a porin with a binding site specific for dicarboxylates which play important key roles in the deficient metabolic and biosynthetic pathways of Borrelia species. The presence of porin P66 has been shown in both Lyme disease and relapsing fever spirochetes. In our study, purified P66 homologues from Lyme disease species B. burgdorferi, B. afzelii and B. garinii and relapsing fever species B. duttonii, B. recurrentis and B. hermsii were compared and their biophysical properties were further characterized in black lipid bilayer assay. Subsequently, the channel diameter of B. burgdorferi P66 was investigated in more detail. For this study, different nonelectrolytes with known hydrodynamic radii were used. This allowed us to determine the effective diameter of the P66 channel lumen. Furthermore, the blockage of the channel after addition of nonelectrolytes revealed seven subconducting states and indicated a heptameric structure of the P66 channel. These results may give more insight into the functional properties of this important porin.

Axonal damage, demyelination and inflammation of the central nervous system are the major pathological features of multiple sclerosis (MS). MS is thought to be due to an abnormal T cell mediated immune response. Oxidative stress plays an important role in the advancement of MS. The reduced glutathione (GSH) has very important role in the management of oxidative stress. In our experiment we used Experimental autoimmune encephalomyelitis (EAE) animal model that mimic human MS and tested the effect of Thymoquinone (TQ) a constituent of oil of Nigella Sativa also known as black seed. Thirty female mice strain C57BL/6J between 6 to 12 weeks of age were placed into 3 groups of 10 and MOG was used subcutaneously (s.c) to induce EAE. Group A, the control group. Group B, received MOG (s.c) and TQ intraperiotoneally (i.p) from day 1 till day 50. Group C, received MOG (s.c) and TQ (i.p) was given on the appearance of first sign and symptoms of Chronic relapsing EAE (CR-EAE). All Mice were examined daily for behavioral deficits and all euthanized and sacrificed on day 50. In this study we found mice belonging to group C (EAE with TQ treatment after the appearance of chronic symptoms) were observed to have the highest mean clinical scores in both the acute and chronic phases of EAE with symptom reduction following the TQ injections. Group B (which received daily TQ injections) had decreased symptoms compared to Group A and C. Glutathione level dropped significantly in the control group (p < 0.05) and increased (p > 0.05) in groups B and C mice who received TQ injections. We also noted that EAE clinical signs correlated well with the extent of perivascular lymphocyte infiltrate compared with normal histology following TQ injections. Our results indicate that TQ, due to its anti-oxidant effects is almost 80% preventive and 50% curative in CR-EAE. These results could assist further studies on the mechanism of the action of TQ in CR-EAE and on the possibility of treating the chronic- relapsing phase of human multiple sclerosis. It seems within the realm of possibility that TQ may be as, if not more, therapeutically efficacious as interferon â and glatiramer acetate.