Lyme disease is the most common tick-borne illness in North America. It can be hard to detect using blood tests, especially in the early stage. Due to the number of significant complications from untreated or undertreated Lyme disease, better methods need to be found to detect the disease. Some surface lipoproteins may be used to detect early disease due to their early expression. Others are maintained for the duration of the infection and can be used to detect chronic Lyme disease. Antibody responses to OspA, OspC and OspE were measured in sera from experimentally infected dogs. The response to OspA was only detectable in a few samples and did not appear to be sensitive for mammalian infections. A protein construct based on multiple epitopes of OspC was used to detect an antibody response starting three weeks after initial infection, and remained detectable for three months. As the response to OspC decreased, OspE could be used to detect an antibody response from three to six months after the initial infection. Because of the strength and the differences in timing of the antibody responses, OspC and OspE could be used to design an accurate blood test for Lyme disease that also indicates early and late stages of infection based on the results.

Laimo boreliozė (LB) yra labiausiai paplitusi erkių platinama liga. Ligos sukėlėjus perneša Ixodes rūšies erkės. Didelį borelijų paplitimą lemia platus rezervuarinių šeimininkų ratas. Šį susirgimą gali sukelti kelios B. burgdorferi s. l. komplekso bakterijų rūšys: B. afzelii, B. garinii, B. burgdorferi sensu stricto. Šiame darbe buvo tirtos 134 Ixodes ricinus erkės nuo 8 elninių gyvūnų (4 stirnų ir 4 elnių) ir 73 erkės nuo žolės (elninių teritorijoje). Naudojant dauginės polimerazės grandininės reakcijos (PGR) medodą, 9 - iose (4,3%) iš jų buvo nustatytas užsikrėtimas Borrelia burgdorferi s. l. patogenais, septyniuose mėginiuose buvo identifikuota B. afzelii, viename - B. garinii rūšis. Viename mėginyje buvo nustatyta Borrelia spp., kuri nebuvo identifikuota. Atlikta B. burgdorferi s. l. sekų analizė pagal ospA geną, naudojantis genų banko duomenimis, atskleidė polimorfizmo lygį tarp trijų, žmogui patogeniškų Borrelia rūšių ir parodė padermių pasiskirstymą skirtingose šalyse bei šeimininkuose. Variabiliausios nustatytos B. garinii ospA geno sekos, rasta 361 variabili vieta (nukleotidų įvairovė π=0,072), aptikta 18 sekų variantų. Mažesnis variabilumas nustatytas B. burgdorferi s. s. sekose – aptiktos 266 variabilios vietos (nukleotidų įvairovė π=0,16) ir 12 sekų variantų bei B. afzelii sekose – aptiktos 257 variabilios vietos (nukleotidų įvairovė π=0,17) ir 4 sekų variantai. Didesni skirtumai buvo identifikuoti tarp B. afzelii ospA geno sekų (vidutinis genetinis atstumas... [toliau žr. visą tekstą] / Lyme borreliosis (LB) is the most common tick - born disease. The pathogens are transmitted by infected ticks belonging to a species of the genus Ixodes. The high prevalence of Borrelia is determined by a wide circle of reservoir hosts. The disease can be caused by a few bacterial species of B. burgdorferi s.l. complex, such as: B. afzelii, B. garinii and B. burgdorferi sensu stricto. 134 Ixodes ricinus ticks have been explored in the thesis, collected from 8 certine animals (4 roe deer and 4 deer) and 73 ticks collected from grass (in the territory of certine animals). Using multiplex polymerase chain reaction (PCR) method, the infection with Borrelia burgdorferi s. l. pathogens was identified in 9 (4.3%), B. afzelii was identified in seven samples, while B. garinii species was found in one of them. Borrelia spp. was found in one sample, which has not been identified. A sequence analysis B. burgdorferi s. l. implemented according to the ospA gene by using a gene bank data revealed level of polymorphism among three Borrelia species pathogenic to human and showed the distribution of strains in the different countries and hosts. The most variable gene sequences are B. garinii ospA. It is found 361 variable place (nucleotide diversity π=0,072), ant detected 18 sequence variants. Less variability found in B. burgdorferi s. s. sequences: 266 variable places are found and 12 sequence variants (nucleotide diversity π=0,16) and 257 variable places are discovered in B. afzelii... [to full text]

The World Health Organization predicts that by the year 2030, road traffic injuries will be one of the top five leading causes of death. Many of these deaths and injuries can be prevented by driving cars properly equipped with state-of-the-art safety and driver assistance systems. Some examples are auto-brake and auto-collision avoidance which are becoming more and more popular on the market today. A recent study by a Swedish insurance company has shown that on roadswith speeds up to 50 km/h an auto-brake system can reduce personal injuries by up to 64 percent. In fact in an estimated 40 percent of crashes, the auto-brake reduced the effects to the degree that no personal injury was sustained. It is imperative that these so called Advanced Driver Assistance Systems, to be really effective, have good situational awareness. It is important that they have adequate information of the vehicle’s immediate surroundings. Where are other cars, pedestrians or motorcycles relative to our own vehicle? How fast are they driving and in which lane? How is our own vehicle driving? Are there objects in the way of our own vehicle’s intended path? These and many more questions can be answered by a properly designed system for situational awareness. In this thesis we design and evaluate, both quantitatively and qualitatively, sensor fusion algorithms for multi-target tracking. We use a combination of camera and radar information to perform fusion and find relevant objects in a cluttered environment. The combination of these two sensors is very interesting because of their complementary attributes. The radar system has high range resolution but poor bearing resolution. The camera system on the other hand has a very high bearing resolution. This is very promising, with the potential to substantially increase the accuracy of the tracking system compared to just using one of the two. We have also designed algorithms for path prediction and a first threat awareness logic which are both qualitively evaluated.

Lyme disease (LD) is the most prevalent vector borne disease is North America with 300,000-600,000 human cases each year. Preventative strategies for LD in humans are poorly developed and largely inadequate. While preventive vaccines for LD are widely used in veterinary medicine, there are no vaccines available for use in humans. The goal of this study was to develop a human vaccine that can elicit antibody responses that kill spirochetes in both the tick and mammalian environments. The approach applied in this study centered on the development of chimeric epitope proteins, referred to as chimeritopes. Chimeritopes consist of a series of epitopes derived from one or more proteins or protein variants. Three chimeritope proteins designated as Chv1, Chv2 and Chv3 were designed. These proteins harbor the same set of 18 linear epitopes derived from 9 different OspC type proteins. They differ in epitope arrangement or by the presence or absence of linkers between specific protein segments. The immunogenicity of each protein was assessed in multiple animal models including mice, rats, and purpose bred beagles. Immunoblot, ELISA, and IFA analyses using sera from immunized animals demonstrated that the Chv proteins elicit IgG responses that recognize a diverse array of OspC type proteins. Anti-Chv and anti-OspA antisera displayed complement dependent bactericidal activity. To assess protective efficacy, purpose bred beagles were immunized with each vaccine formulation and then challenged by infestation with infected ticks. Efficacy was assessed by monitoring seroconversion, cultivation of tissue biopsies, clinical presentation and histopathological analysis of joints and tissues. All dogs vaccinated with the Chv2-OspA combination were fully protected. All dogs in this group were seronegative for LD, biopsy culture negative and did not develop LD associated symptoms including lameness or lesions in tissues or joints. In light of market concerns centered on the use of full length OspA in a human vaccine, epitope mapping was performed to identify a linear epitope that could be employed in development of a possible OspC-OspA chimeritope. A linear epitope, designated as OspA221-240was identified. Antisera to KLH-OspA221-240displayed potent and broad bactericidal activity. Interestingly, the OspA221-240epitope has homology to residues 244 to 263 of OspB suggesting that OspB may also be a potential candidate for inclusion in a human vaccine. This study establishes proof of principle for the use of OspC chimeritopes in LD subunit vaccines and highlights the need to employ a multi-valent, multi-antigen vaccine approach in development of a human LD vaccine.

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Indiana University-Purdue University Indianapolis (IUPUI) / Borrelia burgdorferi, a bacterium which causes Lyme disease, is maintained in nature through a cycle involving two distinct hosts: a tick vector and a mammalian host. To adapt to these two diverse environments, B. burgdorferi undergoes dramatic alterations in its surface lipoprotein. Two essential lipoproteins, outer surface protein A (OspA) and outer surface protein C (OspC), are reciprocally regulated throughout the B. burgdorferi lifecycle. Very little is known about the regulation of OspA. These studies elucidate the regulatory mechanisms controlling the expression of OspA. Various truncations of the ospA promoter were created and then studied in our novel in vitro model of ospA repression or grown within the host-adapted model. A T-Rich region of the ospA promoter was determined to be a cis-element essential for both the full expression and full repression of ospA.