Publications by authors named "Robert Gilmore"

54 Publications

Tofacitinib Is Safe and Effective When Used in Combination With Infliximab for the Management of Refractory Ulcerative Colitis.

Clin Gastroenterol Hepatol 2020 Nov 26. Epub 2020 Nov 26.

Department of Gastroenterology, Austin Health, Department of Medicine, Austin Academic Centre, University of Melbourne, Melbourne, Australia.

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http://dx.doi.org/10.1016/j.cgh.2020.07.046DOI Listing
November 2020

Borrelia miyamotoi strain LB-2001 retains plasmids and infectious phenotype throughout continuous culture passages as evaluated by multiplex PCR.

Ticks Tick Borne Dis 2021 01 6;12(1):101587. Epub 2020 Oct 6.

Bacterial Diseases Branch, Division of Vector-Borne Diseases, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Fort Collins, CO, USA.

Borrelia miyamotoi is a tick-borne spirochete of the relapsing fever borrelia group and an emerging pathogen of public health significance. The genomes of relapsing fever borreliae and Lyme disease borreliae consist of multiple linear and circular plasmids in addition to the chromosome. Previous work with B. burgdorferi sensu lato found diminished infectivity upon continuous in vitro culture passage that was attributable to plasmid loss. The effect of long-term culture passage on B. miyamotoi is not known. We generated a series of plasmid-specific primer sets and developed a multiplex PCR assay to detect the 14 known plasmids of B. miyamotoi North American strains LB-2001 and CT13-2396. We assessed the plasmid content of B. miyamotoi LB-2001 over 64 culture passages spanning 15 months and determined that strain LB-2001 retained all plasmids upon prolonged in vitro cultivation and remained infectious in mice. We also found that strain LB-2001 lacks plasmid lp20-1 which is present in strain CT13-2396. These results suggest that B. miyamotoi remains genetically stable when cultured and passaged in vitro.
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http://dx.doi.org/10.1016/j.ttbdis.2020.101587DOI Listing
January 2021

Cytomegalovirus in inflammatory bowel disease: a clinical approach.

Intern Med J 2020 Oct 3. Epub 2020 Oct 3.

Central Clinical School, Monash University, Melbourne, Victoria, Australia.

Cytomegalovirus (CMV) infection can be a challenging clinical problem in patients with inflammatory bowel disease (IBD), particularly ulcerative colitis. Clinical presentation is difficult to distinguish from an underlying disease flare. A number of diagnostic modalities are now available and when combined can aid clinicians in the identification of patients who are most likely to benefit from antiviral therapy. The aim of this article is to review the available literature and outline a practical approach to the diagnosis and management of CMV in patients with IBD. This article is protected by copyright. All rights reserved.
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http://dx.doi.org/10.1111/imj.15085DOI Listing
October 2020

Characterization of a Borrelia miyamotoi membrane antigen (BmaA) for serodiagnosis of Borrelia miyamotoi disease.

Ticks Tick Borne Dis 2020 09 31;11(5):101476. Epub 2020 May 31.

Bacterial Diseases Branch, Division of Vector-Borne Diseases, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Fort Collins, CO, USA. Electronic address:

Borrelia miyamotoi is a tick-borne pathogen that causes Borrelia miyamotoi disease (BMD), an emerging infectious disease of increasing public health significance. B. miyamotoi is transmitted by the same tick vector (Ixodes spp.) as B. burgdorferi sensu lato (s.l.), the causative agent of Lyme disease, therefore laboratory assays to differentiate BMD from Lyme disease are needed to avoid misdiagnoses and for disease confirmation. We previously performed a global immunoproteomic analysis of the murine host antibody response against B. miyamotoi infection to discover antigens that could serologically distinguish the two infections. An initial assessment identified a putative lipoprotein antigen, here termed BmaA, as a promising candidate to augment current research-based serological assays. In this study, we show that BmaA is an outer surface-associated protein by its susceptibility to protease digestion. Synthesis of BmaA in culture was independent of temperature at either 23 °C or 34 °C. The BmaA gene is present in two identical loci harbored on separate plasmids in North American strains LB-2001 and CT13-2396. bmaA-like sequences are present in other B. miyamotoi strains and relapsing fever borrelia as multicopy genes and as paralogous or orthologous gene families. IgM and IgG antibodies in pooled serum from BMD patients reacted with native BmaA fractionated by 2-dimensional gel electrophoresis and identified by mass spectrometry. IgG against recombinant BmaA was detected in 4 of 5 BMD patient serum samples as compared with 1 of 23 serum samples collected from patients with various stages of Lyme disease. Human anti-B. turicatae serum did not seroreact with recombinant BmaA suggesting a role as a species-specific diagnostic antigen. These results demonstrated that BmaA elicits a human host antibody response during B. miyamotoi infection but not in a tested group of B. burgdorferi-infected Lyme disease patients, thereby providing a potentially useful addition for developing BMD serodiagnostic tests.
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http://dx.doi.org/10.1016/j.ttbdis.2020.101476DOI Listing
September 2020

Using Ustekinumab to Treat Crohn's Disease-Related Orofacial Granulomatosis: Two Birds, One Stone.

Inflamm Bowel Dis 2020 07;26(8):e79-e80

Department of Gastroenterology, Austin Health, Melbourne, Australia.

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http://dx.doi.org/10.1093/ibd/izaa123DOI Listing
July 2020

Evaluation of Patient IgM and IgG Reactivity Against Multiple Antigens for Improvement of Serodiagnostic Testing for Early Lyme Disease.

Front Public Health 2019 5;7:370. Epub 2019 Dec 5.

Division of Vector Borne Diseases, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Fort Collins, CO, United States.

Serologic testing is the standard for laboratory diagnosis and confirmation of Lyme disease. Serodiagnostic assays to detect antibodies against , the agent of Lyme borreliosis, are used for detection of infection. However, serologic testing within the first month of infection is less sensitive as patients' antibody responses continue to develop. Previously, we screened several expressed antigens for candidates that elicit early antibody responses in patients with Stage 1 and 2 Lyme disease. We evaluated patient IgM seroreactivity against 6 antigens and found an increase in sensitivity without compromising specificity when compared to current IgM second-tier immunoblot scoring. In this study, we continued the evaluation using a multi-antigen panel to measure IgM plus IgG seroreactivity in these early Lyme disease patients' serum samples. Using two statistical methods for calculating positivity cutoff values, sensitivity was 70 and 84-87%, for early acute and early convalescent Lyme disease patients, respectively. Specificity was 98-100% for healthy non-endemic control patients, and 96-100% for healthy endemic controls depending on the statistical analysis. We conclude that improved serologic testing for early Lyme disease may be achieved by the addition of multiple borrelial antigens that elicit IgM and IgG antibodies early in infection.
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http://dx.doi.org/10.3389/fpubh.2019.00370DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6906137PMC
December 2019

Immunoproteomic analysis of Borrelia miyamotoi for the identification of serodiagnostic antigens.

Sci Rep 2019 11 14;9(1):16808. Epub 2019 Nov 14.

Bacterial Diseases Branch, Division of Vector-Borne Diseases, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Fort Collins, CO, USA.

The tick-borne spirochete, Borrelia miyamotoi, is an emerging pathogen of public health significance. Current B. miyamotoi serodiagnostic testing depends on reactivity against GlpQ which is not highly sensitive on acute phase serum samples. Additionally, anti-B. miyamotoi antibodies can cross-react with C6 antigen testing for B. burgdorferi, the causative agent of Lyme disease, underscoring the need for improved serological assays that produce accurate diagnostic results. We performed an immunoproteomics analysis of B. miyamotoi proteins to identify novel serodiagnostic antigens. Sera from mice infected with B. miyamotoi by subcutaneous inoculation or tick bite were collected for immunoblotting against B. miyamotoi membrane-associated proteins separated by 2-dimensional electrophoresis (2DE). In total, 88 proteins in 40 2DE immunoreactive spots were identified via mass spectrometry. Multiple variable large proteins (Vlps) and a putative lipoprotein were among those identified and analyzed. Reactivity of anti-B. miyamotoi sera against recombinant Vlps and the putative lipoprotein confirmed their immunogenicity. Mouse anti-B. burgdorferi serum was cross-reactive to all recombinant Vlps, but not against the putative lipoprotein by IgG. Furthermore, antibodies against the recombinant putative lipoprotein were present in serum from a B. miyamotoi-infected human patient, but not a Lyme disease patient. Results presented here provide a comprehensive profile of B. miyamotoi antigens that induce the host immune response and identify a putative lipoprotein as a potentially specific antigen for B. miyamotoi serodetection.
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http://dx.doi.org/10.1038/s41598-019-53248-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6856195PMC
November 2019

Alcohol-induced changes in the gut microbiome and metabolome of rhesus macaques.

Psychopharmacology (Berl) 2019 May 22;236(5):1531-1544. Epub 2019 Mar 22.

Program in Neuroscience, University of Mississippi Medical Center, Jackson, MS, USA.

Rationale: Increasing evidence has demonstrated that changes in the gut microbiome, including those associated with dietary influences, are associated with alterations in many physiological processes. Alcohol consumption is common across human cultures and is likely to have a major effect on the gut microbiome, but there remains a paucity of information on its effects in primates.

Objectives: The effects of chronic alcohol consumption on the primate gut microbiome and metabolome were studied in rhesus macaques that were freely drinking alcohol. The objectives of the study were to determine what changes occurred in the gut microbiome following long-term exposure to alcohol and if these changes were reversible following a period of abstinence.

Methods: Animals consuming alcohol were compared to age-matched controls without access to alcohol and were studied before and after a period of abstinence. Fecal samples from rhesus macaques were used for 16S rRNA sequencing to profile the gut microbiome and for metabolomic profiling using mass spectrometry.

Results: Alcohol consumption resulted in a loss of alpha-diversity in rhesus macaques, though this was partially ameliorated by a period of abstinence. Higher levels of Firmicutes were observed in alcohol-drinking animals at the expense of a number of other microbial taxa, again normalizing in part with a period of abstinence. Metabolomic changes were primarily associated with differences in glycolysis when animals were consuming alcohol and differences in fatty acids when alcohol-drinking animals became abstinent.

Conclusions: The consumption of alcohol has specific effects on the microbiome and metabolome of rhesus macaques independent of secondary influences. Many of these changes are reversed by a relatively short period of abstinence.
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http://dx.doi.org/10.1007/s00213-019-05217-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6613802PMC
May 2019

Evaluation of in vivo expressed Borrelia burgdorferi antigens for improved IgM serodiagnosis of early Lyme disease.

Diagn Microbiol Infect Dis 2019 Mar 3;93(3):196-202. Epub 2018 Oct 3.

Division of Vector Borne Diseases, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Fort Collins, CO, United States of America. Electronic address:

Improved serologic tests are needed for accurate diagnosis and proper treatment of early stage Lyme disease. We evaluated the 3 antigens currently used for 2-tiered IgM immunoblot testing (FlaB, OspC, and BmpA) in combination with 3 additional antigens (BBA65, BBA70, and BBA73) and measured the sensitivity and specificity against a serum repository of positive and negative controls. Using 3 statistical methods for positivity cutoff determinations and scoring criteria, we found increased sensitivities for early Lyme disease when 2 of 6 antigens were positive as compared with the 2 of 3 antigen IgM criteria currently used for second-tier immunoblot scoring. Specificities for negative controls were comparable or superior to using 2 of 3 antigens. These results indicate that IgM sensitivity and specificity of serological testing for Lyme disease in the early stages of illness can be improved by employing antigens that target the initial host antibody responses.
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http://dx.doi.org/10.1016/j.diagmicrobio.2018.09.012DOI Listing
March 2019

Human neuroglial cells internalize Borrelia burgdorferi by coiling phagocytosis mediated by Daam1.

PLoS One 2018 10;13(5):e0197413. Epub 2018 May 10.

Bacterial Diseases Branch, Division of Vector Borne Diseases, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Fort Collins, Colorado, United States of America.

Borrelia burgdorferi, the agent of Lyme borreliosis, can elude hosts' innate and adaptive immunity as part of the course of infection. The ability of B. burgdorferi to invade or be internalized by host cells in vitro has been proposed as a mechanism for the pathogen to evade immune responses or antimicrobials. We have previously shown that B. burgdorferi can be internalized by human neuroglial cells. In this study we demonstrate that these cells take up B. burgdorferi via coiling phagocytosis mediated by the formin, Daam1, a process similarly described for human macrophages. Following coincubation with glial cells, B. burgdorferi was enwrapped by Daam1-enriched coiling pseudopods. Coiling of B. burgdorferi was significantly reduced when neuroglial cells were pretreated with anti-Daam1 antibody indicating the requirement for Daam1 for borrelial phagocytosis. Confocal microscopy showed Daam1 colocalizing to the B. burgdorferi surface suggesting interaction with borrelial membrane protein(s). Using the yeast 2-hybrid system for identifying protein-protein binding, we found that the B. burgdorferi surface lipoprotein, BBA66, bound the FH2 subunit domain of Daam1. Recombinant proteins were used to validate binding by ELISA, pull-down, and co-immunoprecipitation. Evidence for native Daam1 and BBA66 interaction was suggested by colocalization of the proteins in the course of borrelial capture by the Daam1-enriched pseudopodia. Additionally, we found a striking reduction in coiling for a BBA66-deficient mutant strain compared to BBA66-expressing strains. These results show that coiling phagocytosis is a mechanism for borrelial internalization by neuroglial cells mediated by Daam1.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0197413PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5944952PMC
December 2018

Immunization of mice with Borrelia burgdorferi lp54 gene encoded recombinant proteins does not provide protection against tick transmitted infectious challenge.

Vaccine 2017 09 1;35(40):5310-5313. Epub 2017 Sep 1.

Division of Vector Borne Diseases, Centers for Disease Control and Prevention, Fort Collins, CO, USA. Electronic address:

The Borrelia burgdorferi outer surface membrane proteins BBA65, BBA66, BBA69, BBA70, and BBA73 were tested for their ability to confer protection against B. burgdorferi infection challenge. Mice were immunized with recombinant forms of the proteins singly or in combinations. Following initial protein inoculation and booster injections, seroconversion was confirmed prior to B. burgdorferi challenge by tick bite. Despite mice having high antibody titers for each antigen, no significant protections against the challenge infections were observed. These results demonstrate that these recombinant proteins were not protective and reflects the challenges confronted to identify effective novel vaccine candidates for Lyme disease.
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http://dx.doi.org/10.1016/j.vaccine.2017.08.054DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7041491PMC
September 2017

Algorithms for concatenating templates.

Chaos 2016 Mar;26(3):033102

Univ. Bordeaux, LaBRI, UMR5800, F-33400 Talence, France.

We present two algorithms for concatenating two branched manifolds. There exist two common ways to describe representations of branched manifolds with linking matrices. The algorithms provided permit to concatenate branched manifolds with each representation: a linking matrix with an array or only one linking matrix. We also provide algorithms to switch between these two descriptions.
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http://dx.doi.org/10.1063/1.4942799DOI Listing
March 2016

Evaluation of Selected Borrelia burgdorferi lp54 Plasmid-Encoded Gene Products Expressed during Mammalian Infection as Antigens To Improve Serodiagnostic Testing for Early Lyme Disease.

Clin Vaccine Immunol 2015 Nov 16;22(11):1176-86. Epub 2015 Sep 16.

Division of Vector-Borne Diseases, Centers for Disease Control and Prevention, Fort Collins, Colorado, USA

Laboratory testing for the diagnosis of Lyme disease is performed primarily by serologic assays and is accurate for detection beyond the acute stage of the infection. Serodiagnostic assays to detect the early stages of infection, however, are limited in their sensitivity, and improvement is warranted. We analyzed a series of Borrelia burgdorferi proteins known to be induced within feeding ticks and/or during mammalian infection for their utility as serodiagnostic markers against a comprehensive panel of Lyme disease patient serum samples. The antigens were assayed for IgM and IgG reactivity in line immunoblots and separately by enzyme-linked immunosorbent assay (ELISA), with a focus on reactivity against early Lyme disease with erythema migrans (EM), early disseminated Lyme neuroborreliosis, and early Lyme carditis patient serum samples. By IgM immunoblotting, we found that recombinant proteins BBA65, BBA70, and BBA73 reacted with early Lyme EM samples at levels comparable to those of the OspC antigen used in the current IgM blotting criteria. Additionally, these proteins reacted with serum samples from patients with early neuroborreliosis and early carditis, suggesting value in detecting early stages of this disease progression. We also found serological reactivity against recombinant proteins BBA69 and BBA73 with early-Lyme-disease samples using IgG immunoblotting and ELISA. Significantly, some samples that had been scored negative by the Centers for Disease Control and Prevention-recommended 2-tiered testing algorithm demonstrated positive reactivity to one or more of the antigens by IgM/IgG immunoblot and ELISA. These results suggest that incorporating additional in vivo-expressed antigens into the current IgM/IgG immunoblotting tier in a recombinant protein platform assay may improve the performance of early-Lyme-disease serologic testing.
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http://dx.doi.org/10.1128/CVI.00399-15DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4622105PMC
November 2015

Synthesizing Marketing, Community Engagement, and Systems Science Approaches for Advancing Translational Research.

ANS Adv Nurs Sci 2015 Jul-Sep;38(3):227-40

The University of North Carolina at Chapel Hill School of Nursing (Drs Kneipp, Leeman, and Schwartz); Orange County Health Department, Hillsborough, North Carolina (Ms McCall); Gillings School of Global Public Health, The University of North Carolina at Chapel Hill, (Drs Hassmiller-Lich and Schwartz); Research Triangle Institute (RTI) International, Cary, North Carolina (Dr Bobashev and Messrs Riggan and Gil); and Orange County Department of Social Services, Hillsborough, North Carolina (Mr Gilmore).

The adoption and implementation of evidence-based interventions (EBIs) are the goals of translational research; however, potential end-users' perceptions of an EBI value have contributed to low rates of adoption. In this article, we describe our application of emerging dissemination and implementation science theoretical perspectives, community engagement, and systems science principles to develop a novel EBI dissemination approach. Using consumer-driven, graphics-rich simulation, the approach demonstrates predicted implementation effects on health and employment outcomes for socioeconomically disadvantaged women at the local level and is designed to increase adoption interest of county program managers accountable for improving these outcomes in their communities.
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http://dx.doi.org/10.1097/ANS.0000000000000080DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5629963PMC
June 2016

pncA and bptA are not sufficient to complement Ixodes scapularis colonization and persistence by Borrelia burgdorferi in a linear plasmid lp25-deficient background.

Infect Immun 2014 Dec 22;82(12):5110-6. Epub 2014 Sep 22.

Department of Microbial Pathogenesis and Immunology, College of Medicine, Texas A&M Health Science Center, College Station, Texas, USA

The complex segmented genome of Borrelia burgdorferi is comprised of a linear chromosome along with numerous linear and circular plasmids essential for tick and/or mammalian infectivity. The pathogenic necessity for specific borrelial plasmids has been identified; most notably, infections of the tick vector and mammalian host both require linear plasmid 25 (lp25). Genes carried on lp25, specifically bptA and pncA, are postulated to play a role for B. burgdorferi to infect and persist in Ixodes ticks. In this study, we complemented an lp25-deficient borrelial strain with pncA alone or pncA accompanied by bptA to evaluate the ability of the complemented strains to restore larval colonization and persistence through transstadial transmission relative to that of wild-type B. burgdorferi. The acquisition of the complemented strains by tick larvae from infected mice and/or the survival of these strains was significantly decreased when assayed by cultivation and quantitative PCR (qPCR). Only 10% of the pncA-complemented strain organisms were found by culture to survive 17 days following larval feeding, while 45% of the pncA- and bptA-complemented strain organisms survived, with similar results by PCR. However, neither of the complemented B. burgdorferi strains was capable of persisting through the molt to the nymphal stage as analyzed by culture. qPCR analyses of unfed nymphs detected B. burgdorferi genomes in several nymphs at low copy numbers, likely indicating the presence of DNA from dead or dying cells. Overall, the data indicate that pncA and bptA cannot independently support infection, suggesting that lp25 carries additional gene(s) or regulatory elements critical for B. burgdorferi survival and pathogenesis in the Ixodes vector.
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http://dx.doi.org/10.1128/IAI.02613-14DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4249277PMC
December 2014

Nurse scientists overcoming challenges to lead transdisciplinary research teams.

Nurs Outlook 2014 Sep-Oct;62(5):352-61. Epub 2014 May 28.

The Southern Coalition for Social Justice, Durham, NC.

Increasingly, scientific funding agencies are requiring that researchers move toward an integrated, transdisciplinary team science paradigm. Although the barriers to and rewards of conducting this type of research have been discussed in the literature, examples of how nurse investigators have led these teams to reconcile the differences in theoretical, methodological, and/or analytic perspectives that inevitably exist are lacking. In this article, we describe these developmental trajectory challenges through a case study of one transdisciplinary team, focusing on team member characteristics and the leadership tasks associated with successful transdisciplinary science teams in the literature. Specifically, we describe how overcoming these challenges has been essential to examining the complex and potentially cumulative effects that key intersections between legal, social welfare, and labor market systems may have on the health of disadvantaged women. Finally, we discuss this difficult but rewarding work within the context of lessons learned and transdisciplinary team research in relation to the future of nursing science.
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http://dx.doi.org/10.1016/j.outlook.2014.05.002DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5643160PMC
June 2015

Evaluation of the Borrelia burgdorferi BBA64 protein as a protective immunogen in mice.

Clin Vaccine Immunol 2014 Apr 5;21(4):526-33. Epub 2014 Feb 5.

Bacterial Diseases Branch, Division of Vector Borne Diseases, Centers for Disease Control and Prevention, Fort Collins, Colorado, USA.

The Borrelia burgdorferi bba64 gene product is a surface-localized lipoprotein synthesized within mammalian and tick hosts and is involved in vector transmission of disease. These properties suggest that BBA64 may be a vaccine candidate against Lyme borreliosis. In this study, protective immunity against B. burgdorferi challenge was assessed in mice immunized with the BBA64 protein. Mice developed a high-titer antibody response following immunization with soluble recombinant BBA64 but were not protected when challenged by needle inoculation of culture-grown spirochetes. Likewise, mice passively immunized with an anti-BBA64 monoclonal antibody were not protected against needle-inoculated organisms. BBA64-immunized mice were subjected to B. burgdorferi challenge by the natural route of a tick bite, but these trials did not demonstrate significant protective immunity in either outbred or inbred strains of mice. Lipidated recombinant BBA64 produced in Escherichia coli was assessed for possible improved elicitation of a protective immune response. Although inoculation with this antigen produced a high-titer antibody response, the lipidated BBA64 also was unsuccessful in protecting mice from B. burgdorferi challenge by tick bites. Anti-BBA64 antibodies raised in rats eradicated the organisms, as evidenced by in vitro borreliacidal assays, thus demonstrating the potential for BBA64 to be effective as a protective immunogen. However, passive immunization with the same monospecific rat anti-BBA64 polyclonal serum failed to provide protection against tick bite-administered challenge. These results reveal the challenges faced in not only identifying B. burgdorferi proteins with potential protective capability but also in producing recombinant antigens conducive to preventive therapies against Lyme borreliosis.
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http://dx.doi.org/10.1128/CVI.00824-13DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3993113PMC
April 2014

Borrelia burgdorferi bba66 gene inactivation results in attenuated mouse infection by tick transmission.

Infect Immun 2013 Jul 29;81(7):2488-98. Epub 2013 Apr 29.

National Center for Emerging and Zoonotic Infectious Diseases, Division of Vector-Borne Diseases, Bacterial Diseases Branch, Centers for Disease Control and Prevention, Fort Collins, Colorado, USA.

The impact of the Borrelia burgdorferi surface-localized immunogenic lipoprotein BBA66 on vector and host infection was evaluated by inactivating the encoding gene, bba66, and characterizing the mutant phenotype throughout the natural mouse-tick-mouse cycle. The BBA66-deficient mutant isolate, Bb(ΔA66), remained infectious in mice by needle inoculation of cultured organisms, but differences in spirochete burden and pathology in the tibiotarsal joint were observed relative to the parental wild-type (WT) strain. Ixodes scapularis larvae successfully acquired Bb(ΔA66) following feeding on infected mice, and the organisms persisted in these ticks through the molt to nymphs. A series of tick transmission experiments (n = 7) demonstrated that the ability of Bb(ΔA66)-infected nymphs to infect laboratory mice was significantly impaired compared to that of mice fed upon by WT-infected ticks. trans-complementation of Bb(ΔA66) with an intact copy of bba66 restored the WT infectious phenotype in mice via tick transmission. These results suggest a role for BBA66 in facilitating B. burgdorferi dissemination and transmission from the tick vector to the mammalian host as part of the disease process for Lyme borreliosis.
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http://dx.doi.org/10.1128/IAI.00140-13DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3697616PMC
July 2013

Borrelia burgdorferi visualized in Ixodes scapularis tick excrement by immunofluorescence.

Vector Borne Zoonotic Dis 2012 Nov 31;12(11):1000-3. Epub 2012 May 31.

Microbiology and Pathogenesis Activity, Bacterial Diseases Branch, Division of Vector-Borne Diseases, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Fort Collins, Colorado 80521, USA.

The enzootic cycle of Borrelia burgdorferi, the etiologic agent of Lyme disease, involves Ixodes spp. ticks and vertebrates. Resident tick Borrelia, harbored inside the midgut, are eventually expelled with the tick's saliva into the vertebrate host when a tick consumes a blood meal. During this 4- to 5-day feeding period I. scapularis will defecate onto the host's skin. Previously we detected borrelial DNA in tick feces throughout engorgement. In this study we report the microscopic examination for B. burgdorferi in nymphal excrement. Using immunofluorescence assays, we observed Borrelia in all mouse skin and capsule fecal swabs tested, although we could not culture the spirochetes. These results update our previous analysis by revealing that spirochetes can also be visualized in tick excrement. Furthermore, the results emphasize that borrelial contamination by defecation is a possibility, and that caution should be exercised by researchers investigating pathogen/host/vector interactions. The biological significance of the presence of non-culturable Borrelia in tick feces during engorgement is unclear.
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http://dx.doi.org/10.1089/vbz.2011.0922DOI Listing
November 2012

Saliva, salivary gland, and hemolymph collection from Ixodes scapularis ticks.

J Vis Exp 2012 Feb 21(60). Epub 2012 Feb 21.

Microbiology and Pathogenesis Activity, Bacterial Diseases Branch, Division of Vector-Borne Diseases, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention.

Ticks are found worldwide and afflict humans with many tick-borne illnesses. Ticks are vectors for pathogens that cause Lyme disease and tick-borne relapsing fever (Borrelia spp.), Rocky Mountain Spotted fever (Rickettsia rickettsii), ehrlichiosis (Ehrlichia chaffeensis and E. equi), anaplasmosis (Anaplasma phagocytophilum), encephalitis (tick-borne encephalitis virus), babesiosis (Babesia spp.), Colorado tick fever (Coltivirus), and tularemia (Francisella tularensis) (1-8). To be properly transmitted into the host these infectious agents differentially regulate gene expression, interact with tick proteins, and migrate through the tick (3,9-13). For example, the Lyme disease agent, Borrelia burgdorferi, adapts through differential gene expression to the feast and famine stages of the tick's enzootic cycle (14,15). Furthermore, as an Ixodes tick consumes a bloodmeal Borrelia replicate and migrate from the midgut into the hemocoel, where they travel to the salivary glands and are transmitted into the host with the expelled saliva (9,16-19). As a tick feeds the host typically responds with a strong hemostatic and innate immune response (11,13,20-22). Despite these host responses, I. scapularis can feed for several days because tick saliva contains proteins that are immunomodulatory, lytic agents, anticoagulants, and fibrinolysins to aid the tick feeding (3,11,20,21,23). The immunomodulatory activities possessed by tick saliva or salivary gland extract (SGE) facilitate transmission, proliferation, and dissemination of numerous tick-borne pathogens (3,20,24-27). To further understand how tick-borne infectious agents cause disease it is essential to dissect actively feeding ticks and collect tick saliva. This video protocol demonstrates dissection techniques for the collection of hemolymph and the removal of salivary glands from actively feeding I. scapularis nymphs after 48 and 72 hours post mouse placement. We also demonstrate saliva collection from an adult female I. scapularis tick.
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http://dx.doi.org/10.3791/3894DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3912584PMC
February 2012

Analysis of Borrelia burgdorferi Surface Proteins as Determinants in Establishing Host Cell Interactions.

Front Microbiol 2011 1;2:141. Epub 2011 Jul 1.

Bacterial Diseases Branch, Division of Vector Borne Diseases, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention Fort Collins, CO, USA.

Borrelia burgdorferi infection causes Lyme borreliosis in humans, a condition which can involve a systemic spread of the organism to colonize various tissues and organs. If the infection is left untreated by antimicrobials, it can lead to manifestations including, arthritis, carditis, and/or neurological problems. Identification and characterization of B. burgdorferi outer membrane proteins that facilitate cellular attachment and invasion to establish infection continue to be investigated. In this study, we sought to further define putative cell binding properties of surface-exposed B. burgdorferi proteins by observing whether cellular adherence could be blocked by antibodies. B. burgdorferi mixed separately with monoclonal antibodies (mAbs) against outer surface protein (Osp) A, OspC, decorin-binding protein (Dbp) A, BBA64, and RevA antigens were incubated with human umbilical vein endothelial cells (HUVEC) and human neuroglial cells (H4). B. burgdorferi treated with anti-OspA, -DbpA, and -BBA64 mAbs showed a significant decrease in cellular association compared to controls, whereas B. burgdorferi treated with anti-OspC and anti-RevA showed no reduction in cellular attachment. Additionally, temporal transcriptional analyses revealed upregulated expression of bba64, ospA, and dbpA during coincubation with cells. Together, the data provide evidence that OspA, DbpA, and BBA64 function in host cell adherence and infection mechanisms.
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http://dx.doi.org/10.3389/fmicb.2011.00141DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3129520PMC
October 2012

Functional analysis of the Borrelia burgdorferi bba64 gene product in murine infection via tick infestation.

PLoS One 2011 May 3;6(5):e19536. Epub 2011 May 3.

Microbiology and Pathogenesis Activity, Division of Vector-Borne Diseases, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Fort Collins, Colorado, USA.

Borrelia burgdorferi, the causative agent of Lyme borreliosis, is transmitted to humans from the bite of Ixodes spp. ticks. During the borrelial tick-to-mammal life cycle, B. burgdorferi must adapt to many environmental changes by regulating several genes, including bba64. Our laboratory recently demonstrated that the bba64 gene product is necessary for mouse infectivity when B. burgdorferi is transmitted by an infected tick bite, but not via needle inoculation. In this study we investigated the phenotypic properties of a bba64 mutant strain, including 1) replication during tick engorgement, 2) migration into the nymphal salivary glands, 3) host transmission, and 4) susceptibility to the MyD88-dependent innate immune response. Results revealed that the bba64 mutant's attenuated infectivity by tick bite was not due to a growth defect inside an actively feeding nymphal tick, or failure to invade the salivary glands. These findings suggested there was either a lack of spirochete transmission to the host dermis or increased susceptibility to the host's innate immune response. Further experiments showed the bba64 mutant was not culturable from mouse skin taken at the nymphal bite site and was unable to establish infection in MyD88-deficient mice via tick infestation. Collectively, the results of this study indicate that BBA64 functions at the salivary gland-to-host delivery interface of vector transmission and is not involved in resistance to MyD88-mediated innate immunity.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0019536PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3086921PMC
May 2011

Detection of Borrelia burgdorferi DNA in tick feces provides evidence for organism shedding during vector feeding.

Vector Borne Zoonotic Dis 2011 Mar 6;11(3):197-200. Epub 2010 Oct 6.

Microbiology and Pathogenesis Activity, Division of Vector-Borne Diseases, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Fort Collins, CO 80521, USA.

Borrelia burgdorferi, the bacterium that causes Lyme disease, is transmitted to a susceptible host by Ixodes spp. tick bites. However, there is uncertainty whether B. burgdorferi are shed from ticks by the fecal route. In this study, B. burgdorferi-infected ticks were fed on mice while confined to a certain area of the skin by a capsule. During and after feeding, tick feces were collected and placed in Barbour-Stoenner-Kelley (BSK)-II media for cultivation and in sterile water for polymerase chain reaction (PCR) analysis. Although none of the tested samples were culture positive for B. burgdorferi, all but one of the fecal DNA samples from infected ticks were PCR positive. These results indicated that B. burgdorferi were shed from feeding ticks during defecation and suggest that the spirochetes did not remain viable once exposed to the outside environment. This finding has important ramifications for investigators interpreting B. burgdorferi-specific PCR results when conducting tick transmission experiments.
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http://dx.doi.org/10.1089/vbz.2010.0149DOI Listing
March 2011

The bba64 gene of Borrelia burgdorferi, the Lyme disease agent, is critical for mammalian infection via tick bite transmission.

Proc Natl Acad Sci U S A 2010 Apr 5;107(16):7515-20. Epub 2010 Apr 5.

Bacterial Diseases Branch, Division of Vector-Borne Infectious Diseases, Centers for Disease Control and Prevention, Fort Collins, CO 80521, USA.

The spirochetal agent of Lyme disease, Borrelia burgdorferi, is transmitted by bites of Ixodes ticks to mammalian reservoir hosts and humans. The mechanism(s) by which the organism is trafficked from vector to host is poorly understood. In this study, we demonstrate that a B. burgdorferi mutant strain deficient in the synthesis of the bba64 gene product was incapable of infecting mice via tick bite even though the mutant was (i) infectious in mice when introduced by needle inoculation, (ii) acquired by larval ticks feeding on infected mice, and (iii) able to persist through tick molting stages. This finding of a B. burgdorferi gene required for pathogen transfer and/or survival from the tick to the susceptible host represents an important breakthrough toward understanding transmission mechanisms involved for the Lyme disease agent.
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http://dx.doi.org/10.1073/pnas.1000268107DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2867675PMC
April 2010

Biological algorithm for data reconstruction.

Phys Rev E Stat Nonlin Soft Matter Phys 2010 Mar 24;81(3 Pt 2):036217. Epub 2010 Mar 24.

Physics Department, Drexel University, Philadelphia, Pennsylvania 19104, USA.

An algorithm inspired by Genome sequencing is proposed which "reconstructs" a single long trajectory of a dynamical system from many short trajectories. This procedure is useful in situations when many data sets are available but each is insufficiently long to apply a meaningful analysis directly. The algorithm is applied to the Rössler and Lorenz dynamical systems as well as to experimental data taken from the Belousov-Zhabotinskii chemical reaction. Topological information was reliably extracted from each system and geometrical and dynamical measures were computed.
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http://dx.doi.org/10.1103/PhysRevE.81.036217DOI Listing
March 2010

Borrelia burgdorferi expression of the bba64, bba65, bba66, and bba73 genes in tissues during persistent infection in mice.

Microb Pathog 2008 Nov-Dec;45(5-6):355-60. Epub 2008 Sep 20.

Division of Vector-Borne Infectious Diseases, Centers for Disease Control and Prevention, 3150 Rampart Rd, Fort Collins, CO 80521, USA.

Borrelia burgdorferi, the etiological agent of Lyme disease in humans, is vectored between mammalian hosts in nature by Ixodes ticks. The organism adapts to diverse environments encountered throughout the enzootic cycle by differentially expressing essential gene products to survive the specialized conditions, whether in ticks or warm-blooded hosts. However, little is known regarding the identity and/or function of B. burgdorferi genes expressed during colonization of tissues during mammalian infection. Experimental evidence has shown that a group of genes (formerly classified as paralogous gene family 54) contiguously localized on the 54-kilobase linear plasmid of B. burgdorferi, are among the most highly regulated by in vitro conditions resembling mammalian infection. In this study, we employed quantitative reverse transcription-PCR to measure temporal gene expression of a subset of this B. burgdorferi gene family (bba64, bba65, bba66, and bba73) in tissues during chronic murine infection. The goal was to gain insight into the role of these genes in infectivity and pathogenesis by identifying when the genes are induced and whether they are expressed in specific target tissues. B. burgdorferi bba64, bba65, bba66, and bba73 expression was measured from infected mouse tissues relative to expression in in vitro culture conditions at specific times post-infection. bba64 expression was highly upregulated in bladder, heart, and spleen tissues throughout the infection period, contrasting with the sharp downregulation previously observed in ear tissues. bba65, bba66, and bba73 demonstrated upregulated differential expression in various tissues over 1 year post-infection. These results suggest an essential role for these genes in borrelial survival, persistence, and/or pathogenesis.
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http://dx.doi.org/10.1016/j.micpath.2008.08.006DOI Listing
February 2009

From quasiperiodicity to toroidal chaos: analogy between the Curry-Yorke map and the van der Pol system.

Phys Rev E Stat Nonlin Soft Matter Phys 2008 Apr 7;77(4 Pt 2):046203. Epub 2008 Apr 7.

CORIA UMR 6614-Université de Rouen, BP 12, F-76801 Saint-Etienne du Rouvray cedex, France.

The van der Pol attractor exhibits a wide variety of behavior depending on the control parameter values: limit cycles, quasiperiodic motion on a torus, mode locking, period doubling, banded chaos, boundary crises, torus wrinkling, breakup of a torus, and toroidal chaos. The organization of these phenomena with respect to each other is well described by studying a partition of the control parameter plane of the Curry-Yorke map.
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http://dx.doi.org/10.1103/PhysRevE.77.046203DOI Listing
April 2008

Borrelia burgdorferi surface-localized proteins expressed during persistent murine infection are conserved among diverse Borrelia spp.

Infect Immun 2008 Jun 7;76(6):2498-511. Epub 2008 Apr 7.

University of Pittsburgh School of Medicine, Department of Microbiology and Molecular Genetics, 200 Lothrop Street, Pittsburgh, PA 15261, USA.

Borrelia burgdorferi, the causative agent of Lyme disease in the United States, regulates numerous genes encoding lipoproteins on linear plasmid 54 in response to environmental cues. We analyzed a subset of these genes/proteins that were historically categorized as paralogous gene family 54 (BBA64, BBA65, BBA66, BBA68, BBA69, BBA70, BBA71, and BBA73) and found that the expression of several genes was influenced by the sigma(N)-sigma(S) regulatory cascade at the level of transcription and protein synthesis. Moreover, we established in this and a previous study that BBA65, BBA66, BBA69, BBA71, and BBA73 are temporally expressed during persistent infection of immunocompetent mice, as determined by quantitative real time-PCR of ear tissue, by enzyme-linked immunosorbent assay, and by immunoblotting. Correspondingly, BBA65, BBA66, BBA71, and BBA73 proteins were detectable in infectious B. burgdorferi B31 isolates but undetectable in noninfectious isolates. BBA65, BBA66, BBA71, and BBA73 proteins were also found to partition into the Triton X-114 detergent phase and were sensitive to protease treatment of intact cells, indicating that they are membrane associated and surface localized. Lastly, Southern blotting and PCR with specific gene primer/probes for BBA64, BBA65, BBA66, BBA71, and BBA73 suggest that many of these genes are conserved among the B. burgdorferi sensu lato isolates and the relapsing-fever Borrelia species. Together, the data presented suggest that these genes may play a part in Borrelia infection and/or pathogenicity that could extend beyond the sensu lato group.
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http://dx.doi.org/10.1128/IAI.01583-07DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2423095PMC
June 2008

Peeling bifurcations of toroidal chaotic attractors.

Phys Rev E Stat Nonlin Soft Matter Phys 2007 Dec 11;76(6 Pt 2):066204. Epub 2007 Dec 11.

CORIA UMR 6614--Université de Rouen, Av. de l'Université, Boîte Postale 12, F-76801 Saint-Etienne du Rouvray cedex, France.

Chaotic attractors with toroidal topology (van der Pol attractor) have counterparts with symmetry that exhibit unfamiliar phenomena. We investigate double covers of toroidal attractors, discuss changes in their morphology under correlated peeling bifurcations, describe their topological structures and the changes undergone as a symmetry axis crosses the original attractor, and indicate how the symbol name of a trajectory in the original lifts to one in the cover. Covering orbits are described using a powerful synthesis of kneading theory with refinements of the circle map. These methods are applied to a simple version of the van der Pol oscillator.
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http://dx.doi.org/10.1103/PhysRevE.76.066204DOI Listing
December 2007