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Current Thesis Projects

 

Attacking Tick-borne Pathogens

Cheryl Bandoski

Thesis Project title: Identification and genotypic analysis of pathogens present in Morgellons disease patient skin samples.

Description: Morgellons disease (MD) is a self-diagnosed, pruritic skin condition characterized by microscopic fibers below unbroken skin or protruding from slow healing lesions.  In addition, patients experience crawling, stinging, and biting sensations under the skin.  It has become evident that the dermal filaments are associated with multisystemic symptoms resembling Lyme disease such as fatigue, cognitive decline, and joint pain. Many MD patients have positive serology to Borrelia burgdorferi and the finding of Borrelia burgdorferi in the skin of MD patients has suggested the disease is a manifestation of Lyme disease.  This view is highly contested, as MD is considered a delusional disease by the medical community. This study was undertaken to identify any potentially infectious organisms present in MD patient skin lesions with the goal of gaining an understanding of what may lead to lesion and fiber formation.  A metagenomic sequencing approach was taken for the identification of pathogenic organisms.  Potentially infectious organisms present in several patient samples were further investigated by multilocus sequence typing using PCR and were visualized in tissue samples by immunohistochemistry and in situ hybridization.

Priyanka Theophilus

Thesis project title: The Stevia Story.

Description: The spirochete Borrelia burgdorferi is the etiological agent of Lyme borreliosis. Administering antibiotics is the frontline treatment for this disease; however, Borrelia is highly resistant to antibiotics. It has been proposed that the resistance might be due to the formation of different defensive forms of Borrelia, namely cysts and the recently proposed biofilm form. Recent studies on antibiotic resistant pathogens show that alternative clinical treatment option involving medicinal antimicrobial agents could have significant antimicrobial, antiprotozoal, antiviral properties and in addition to that they are proven to be nontoxic and can be taken safely for longer period of time. Previously we have reported that Samento and Banderol agents have significant antimicrobial activity against Lyme bacteria. The purpose of this study is to investigate the in vitro effectiveness of several other Peruvian antimicrobial agents on Borrelia and to study the synergistic effect of natural antimicrobial agents with antibiotics in an attempt to eliminate the different morphological forms of Borrelia. In consequence, this might provide a more effective therapeutic option for Lyme disease patients.

Shafiq Shaikh

Thesis project title: How Does Borrelia Invade Human Blood Cells?

Description: I am currently working on co-culturing Borrelia burgdorferi spirochetes with blood. Borrelia burgdorferi is the Lyme disease causing spirochete that is transmitted through Ixodes spp tick using blood as a medium of transmission. The mechanism Borrelia burgdorferi spirochete uses to persist with the blood and its use as a mode of transmission between the vector and the host is unknown. Hence it is my goal to explore the invasion of several kinds of blood cells by Borrelia burgdorferi spirochetes as a possible model of infection by the spirochete inside the human body. 

Kayla Soccaras

Thesis project title: The Affects of Co-Culturing on Borrelia.

Description: Infectious diseases such as Borreliosis, commonly known as Lyme disease have become increasingly common in endemic areas throughout North America, Europe, and Asia. The cause of this Borreliosis is Borrelia burgdorferi, a spirocheteal bacterium capable of altering its morphology and physiology in accordance to its environmental surroundings. The full extent of the impact of Borrelia burgdorferi on mammalian cells is largely unknown due to the limitations in diagnostic tools and genetic variation in individuals afflicted with the Borreliosis. It is hypothesized that when co-cultured with Borrelia burgdorferi, mammalian cells will undergo alterations in morphology and physiology. Thus in my research the differences in morphology, physiology, and gene expression of mammalian cells when cultured normally and when cultured in the presence of an infectious agent such as North American or European strains of Borrelia burgdorferi will be investigated.

Renée Heroux

Research project title: Can Lyme Disease be Sexually Transmitted?

Description: Lyme disease was not proven to be sexually transmitted despite the fact that a closely related spirochete called Treponema can cause a well-known sexually transmitted disease called syphilis. My hypothesis in this research project that Borrelia DNA can be found in reproductive fluids from Lyme disease patients. I will be obtaining fixed blinded clinical samples from Lyme disease patients as well as from healthy controls and I will use PCR, direct sequencing technologies to detect Borrelia DNA in these samples and Mega5 phylogenetic program to analyze the obtained sequences. If we can prove that Borrelia DNAs exist in these clinical samples it will warrant a further investigation for the possibility that Lyme disease is sexually transmitted. 

 Katherine R Filush

Thesis project title: In vivo la Revolution.

Description: Lyme disease is an ever-growing concern with many new cases confirmed each year. Although Lyme disease is currently treated with a course of antibiotics many patients continue to experience symptoms. The possibility for Borrelia to become resistant to antibiotics requires us to reevaluate our current understanding of Borrelia and investigate the potential of other forms such as biofilms, which have been seen in vitro. Biofilms and round body forms have served as a defense against environmental stress for many bacteria. If Borrelia does exist in these forms in vitro and in vivo than this could be the possible cause of resistance to the current antibiotic treatment. The purpose of my project is to expand the current knowledge of possible multiple morphological forms of Borrelia in different types of tissue could shift  our understanding of Borrelia. 

Kunthavai Balasubramanian

Thesis project title: Evidence for presence of Borrelia burgdorferi Biofilm structures in Erythema Chronic Migrans (Bull’s Eye Rash) Infected Human Skin Tissues

Description: Lyme borreliosis is an infectious disease caused by the bacterium Borrelia burgdorferi. A tick bite and infection with Borrelia burgdorferi leads to a skin lesion called Bull’s eye rash or Erythema Chronicum Migrans rash. We recently reported a novel form for Borrelia biofilm, a complex aggregate which can adhere to biotic and abiotic surfaces and thereby, provides protection from hostile environments or antibiotic therapy. The presence of Borrelia has not been demonstrated in human infected tissues despite the fact that biofilm structure can provide a protective structure for Borrelia.  In this study, we show evidence for the presence of Borrelia biofilm in vivo using human skin tissues from patients presenting Erythema Chronicum Migrans skin rashes. Using immunohistochemical staining and silver staining methods, we report the presence of of Borrelia as well as alginate, a marker for the presence of biofilm, in the human infected tissues.

Jasmin Maghsoudlou

Thesis project title: The Connection Between Borrelia and Neurological Diseases

Description: Lyme disease is caused by the bacterium Borrelia burgdorferi. Infection with this bacterium can lead to neurological manifestations. Lyme neuroborreliosis (LNB) commonly involves the cranial nerves, spinal nerve roots, and meninges. LNB can mimic strokes, brain tumors, meningoencephalitis, autoimmune diseases, multiple sclerosis or Alzheimer’s disease. Spirochetes could directly and indirectly cause brain lesions. The cause of Alzheimer’s disease, multiple sclerosis (MS), and many other neurological disorders has long been unknown. Previous studies have observed a spirochetal presence in MS and Alzheimer’s disease tissues. The similarities between LNB and many neurological diseases could be explained by a spirochetal presence. I hypothesize that the presence of Borrelia burgdorferi is detectable in human neurological diseased brain tissue. The presence of Borrelia burgdorferi will be analyzed and studied by Borrelia burgdorferi specific immunohistochemistry, in situ hybridization, standard silver staining methods as well as polymerase chain reaction (PCR) techniques in human neurological diseased brain tissue. This presence will be visualized and characterized through atomic force, differential interference contrast (DIC) and fluorescent microscopy methods.

Maria Jacintha Victoria

Thesis Project title: Finding Alternative Treatments for Lyme Disease

Description:  Lyme borreliosis is a tick borne infective disease caused by Borrelia burgdorferi. Although antibiotics are the preferred course of treatment, the disease becomes increasingly antibiotic resistant. This resistance has been attributed to the biofilm, a defensive morphological form. There is an urgent need to develop novel therapeutic strategies to eliminate Borrelia biofilm forms. Previous studies have shown that treating biofilms with sugar derivatives increases their metabolic activity which makes them susceptible to antibiotic treatment. In addition, enzymes have also shown to reduce viability of biofilms. In my research, I treated Borrelia burgdorferi, grown previously on solid support, with sugars and enzymes in combination with antibiotics.  Our result shows that certain enzymes and sugar derivatives in combination with gentamycin and doxycycline antibiotics, dramatically decreases the viability of Borrelia burgdorferi biofilm. As sugar derivatives and enzymes are non-toxic agents, it could be added to any antimicrobial therapy to enhance the efficiency of antibiotics.