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Past Publications for LDA-Funded Research 2

The LDA has a long history of funding/supporting research projects that lead to real results. Below is the list of links to peer-reviewed articles that acknowledge LDA’s funding/support. Click on the publication link (or just scroll down to it) to see the authors, which authors LDA provided the funding for (green), and the abstract and a link to the publication itself when possible. When a journal has multiple publications, click on the year in which you are interested.

60 Publications

Journal of Medical Entomology 2022
Healthcare (Basel) 2022, 2018
The Journal of Nervous and Mental Disease 2022
Journal of Vector Ecology 2021
Frontiers in Neurology 2021Healthcare
(2) Frontiers in Medicine 2020, 2019
(2) Antibiotics 2020, 2017
Brain, Behavior, & Immunity – Health 2020
Meta Gene 2019
Ticks and Tick-Borne Diseases 2018
Archives of Clinical Neuropsychology 2018
(2) Psychosomatics 2018, 2013
Bio-Protocol 2017

(5) PLOS One 2017, 2012 (2), 2011, 2010
The FEBS Journal 2017
ACS Chemical Biology 2017
Biochemistry 2017
(2) Frontiers in Microbiology October 2016May 2016
Park Science (NPS, Department of Interior) 2016
FEMS Microbiology Letters 2015
Clinical Infectious Disease 2014
Veterinary Sciences 2014
International Journal of Medical Sciences 2013
Northeastern Naturalist, 2013

(3)Journal of Neuropsychiatry & Clinical Neurosciences 2013, 2003, 2001
Open Neurology Journal 2012
Journal of Bacteriology 2011 Genetics 2011>
Journal of Medical Entomology 2010>
Neurobiology of Disease 2010
Archives General Psychiatry 2009
Gene 2009
(2) Neurology 2008, 1999
Emerging Infectious Diseases July 2008
Minerva Medica October 2008
Microbial Pathogenesis 2008>
Journal of International Neuropsychological Soc. 2006
Infection & Immunity 2006
Journal of Clinical Microbiology 2005
Expert Review of Anti-Infective Therapy 2004
The Proceedings of National Academy of Science 2004
(2) JSTBD 2002, 1999
Medscape Infectious Diseases April 2000>
Journal of American Medical Association (JAMA) 1999>
Psychiatric Clinics of North America 1998
(2) Infection 1998, 1996


Click here to see a sample of conference presentations from researchers who have received LDA funding for their work.

Conference Presentations Resulting from LDA-Funding


Journal Articles Below

60. Journal of Medical Entomology, August 2022

Tick-Borne Pathogens in Questing Blacklegged Ticks (Acari: Ixodidae) From Pike County, Pennsylvania 3 August 2022

https://academic.oup.com/jme/advance-article/doi/10.1093/jme/tjac107/6653590

Sarah Schwartz,1, Elizabeth Calvente,1, Emily Rollinson,2, Destiny Sample Koon Koon,1, and Nicole Chinnici1,3

1Dr. Jane Huffman Wildlife Genetics Institute, East Stroudsburg University of Pennsylvania, 562 Independence Road, Suite 114,East Stroudsburg, PA 18301, USA

2East Stroudsburg University, 200 Prospect Street, East Stroudsburg, PA 18301, USA
3
Corresponding author, e-mail: nchinici@esu.edu

Abstract
: Active surveillance was conducted by collecting questing ticks from vegetation through a 2-yr survey in Pike County, Pennsylvania. Over a thousand blacklegged ticks (Ixodes scapularis Say) and American dog ticks (Dermacentor variabilis Say) were collected. A single specimen of the following species was collected: lone star tick (Amblyomma americanum L.), rabbit tick (Haemaphysalis leporispalustris Packard), and an Asian longhorned tick (Haemaphysalis longicornis Neumann). This study represents the largest county-wide study in Pennsylvania, surveying 988 questing I. scapularis adult and nymphs. Molecular detection of five distinct tick-borne pathogens was screened through real-time PCR at a single tick resolution. Respectively, the overall 2-yr adult and nymph prevalence were highest with Borrelia burgdorferi (Spirochaetales: Spirochaetacceae) (45.99%, 18.94%), Anaplasma phagocytophilum (Rickettsiales: Anaplasmataceae) (12.29%, 7.95%) where the
variant-ha (8.29%, 3.03%) was overall more prevalent than the variant-v1 (2.49%, 4.17%),
Babesia microti (Piroplasmida: Babesiidae) (4.97%, 5.30%), Borrelia miyamotoi (Spirochaetales: Spirochaetaceae) (1.38%, 1.89%), and Powassan virus lineage II [POWV]/deer tick virus (DTV) (2.07%, 0.76%). Adult and nymph coinfection prevalence of B. burgdorferi and B. microti (3.03%, 4.97%) and adult coinfection of B. burgdorferi and A. phagocytophilum or A. phagocytophilum and B. microti were significantly higher than the independent infection rate expected naturally. This study highlights the urgency to conduct diverse surveillance studies with large sample sizes to better understand the human risk for tick-borne diseases within small geographical area.

59. Healthcare (Basel) 2022 July 22

Kundalini Yoga for Post-Treatment Lyme Disease: A Preliminary Randomized Study

https://www.mdpi.com/2227-9032/10/7/1314/htm
Lilly Murray 1,2,* , Charles Alexander 3, Clair Bennett 1,2, Mara Kuvaldina 1,2, Gurucharan Khalsa 4 and Brian Fallon 1,2,*

1 Lyme & Tick-Borne Diseases Research Center, Department of Psychiatry, Columbia University Irving Medical Center, New York, NY 10032, USA; cb3492@cumc.columbia.edu (C.B.); mk4480@cumc.columbia.edu (M.K.)

2 New York State Psychiatric Institute, New York, NY 10032, USA

3 Private Practice, Southport, CT 06890, USA; chipalex13@aol.com

4 Fish Interfaith Center, Chapman University, Orange, CA 92866, USA; khalsa@chapman.edu

* Correspondence: lm3448@cumc.columbia.edu (L.M.); baf1@cumc.columbia.edu (B.F.)

Abstract: This study examined the adherence to and the potential benefit of Kundalini yoga (KY) for post-treatment Lyme disease syndrome (PTLDS). Participants were randomly assigned to 8 weeks of a KY small-group intervention or a waitlist control (WLC). Adherence was measured as attendance at KY group sessions. Primary outcomes assessed pain, pain interference, fatigue, and global health. Secondary outcomes assessed multisystem symptom burden, mood, sleep, physical and social functioning, cognition, and mindfulness. Linear mixed models were used to test changes in outcomes over time as a function of group assignment; intercepts for participants were modeled as random effects. Although the target sample size was 40 participants, the study concluded with 29 participants due to recruitment challenges. No KY participants dropped out of the study, and participants attended 75% of group sessions on average, but WLC retention was poor (57%). Regarding primary outcomes, there was no significant interaction between group and time. Regarding secondary outcomes, there was a significant interaction between group and time for multisystem symptom burden (p < 0.05) and cognition (p < 0.01); KY participants reported improved multisystem symptom burden and cognition over the course of the study compared to WLC participants. To enhance recruitment and retention, future trials may consider expanding geographic access and including supportive procedures for WLC participants. This preliminary study supports the need for a larger study to determine if KY reduces multisystem symptom burden and enhances cognition among people with PTLDS

58. The Journal of Nervous and Mental Disease: May 2022 – Volume 210 – Issue 5
Persistent Symptoms, Lyme Disease, and Prior Trauma
https://journals.lww.com/jonmd/Abstract/2022/05000/Persistent_Symptoms,_Lyme_Disease,_and_Prior.7.aspx
Mustafiz, Fayel BA∗,†; Moeller, James PhD∗,†; Kuvaldina, Maria PhD∗,†;
Bennett, Clair DPsych (Clin)∗,†; Fallon, Brian A. MD∗,†
∗Columbia Psychiatry, Columbia University Irving Medical Center
†New York Psychiatric Institute, New York, New York.

Correspondence: Fayel Mustafiz, BA, 1051 Riverside Drive, Unit 69, New
York, NY 10032. E-mail: mustafiz.fayel@gmail.com.

Abstract: One prior study suggests that traumatic events before Lyme disease play an important role in symptom severity. We examined this hypothesis among 60 individuals with persistent symptoms after Lyme disease using validated measures of trauma history, mental and physical symptoms, and functional status. Analysis of variance with Tukey-Kramer multiple comparisons test revealed that a greater number of traumatic events were significantly associated with greater symptom severity on the scales of mood (stress, depression, and anxiety), cognition, multisystem symptom burden, and functional status (mental and physical), but not on measures of pain and fatigue. The effect sizes—meaningful but not large (0.17–0.29)—were mostly produced by comparison with individuals reporting multiple prior traumatic events, representing half of the posttreatment Lyme disease syndrome (PTLDS) group. In conclusion, although PTLDS may be exacerbated by past trauma, trauma plays a role in only a subgroup of PTLDS. Whether addressing prior trauma can improve outcomes in this subgroup requires study.


57. Journal of Vector Ecology, Vol. 46, Issue, Nov 2021
Establishment of Amblyomma americanum populations and new records of Borrelia burgdorferi-infected Ixodes scapularis in South Dakota https://bioone.org/journals/journal-of-vector-ecology/volume-46/issue-2/1081-1710-46.2.143/Establishment-of-Amblyomma-americanum-populations-and-new-records-of-Borrelia/10.52707/1081-1710-46.2.143.short

Holly Black 1,2, Rashaun Potts 2, Jayden Fiechtner 2, Jose E. Pietri 2, Hugh B. Britten 1

1 Department of Biology, University of South Dakota, Vermillion, SD, U.S.A.
2 Sanford School of Medicine, University of South Dakota, Vermillion, SD, U.S.A.

Abstract: Tick-borne diseases are an emerging public health threat in the United States, but surveillance is lacking in some regions. To advance current knowledge of the ecology of ticks and tick-borne diseases in South Dakota, we conducted a survey in the summer of 2019, focusing on the eastern counties of the state. We collected and identified 266 ticks and a subset were tested for the presence of Borrelia burgdorferi by polymerase chain reaction (PCR). Dermacentor variabilis, a ubiquitous species in the state, was the most commonly identified tick, present in all counties surveyed. However, we also identified 15 Amblyomma americanum from three different locations, providing the first evidence of established populations in the state and expanding the range of this species. In addition, we identified 22 Ixodes scapularis from five different locations, confirming a previous report of an established population in the state. Two adult I. scapularis from two different sites were found to harbor B. burgdorferi, including an individual from Lincoln County, suggesting the ongoing presence of the pathogen in tick populations in the state and representing its southwestern-most detection in the midwest United States. These findings provide important information for assessing and monitoring the public health risk from tick-borne diseases in an area where surveillance is lacking.


56. Frontiers in Neurology, May 10, 2021
Detecting Borrelia Spirochetes: A Case Study With Validation Among Autopsy Specimens https://www.frontiersin.org/articles/10.3389/fneur.2021.628045/full

Shiva Kumar Goud Gadila1, Gorazd Rosoklija2,3, Andrew J. Dwork2,3,4,5, Brian A. Fallon2* and Monica E. Embers1*

1 Division of Immunology, Tulane National Primate Research Center, Tulane University Health Sciences, Covington, LA, United States
2 Department of Psychiatry, Columbia University, New York, NY, United States
3 Division of Molecular Imaging and Neuropathology, New York State Psychiatric Institute, New York, NY, United States
4 Macedonian Academy of Sciences and Arts, Skopje, Macedonia
5 Department of Pathology and Cell Biology, Columbia University, New York, NY, United States

*Correspondence: Brian A. Fallon, baf1@cumc.columbia.edu; Monica E. Embers, members@tulane.edu

Abstract: The complex etiology of neurodegenerative disease has prompted studies on multiple mechanisms including genetic predisposition, brain biochemistry, immunological responses, and microbial insult. In particular, Lyme disease is often associated with neurocognitive impairment with variable manifestations between patients. We sought to develop methods to reliably detect Borrelia burgdorferi, the spirochete bacteria responsible for Lyme disease, in autopsy specimens of patients with a history of neurocognitive disease. In this report, we describe the use of multiple molecular detection techniques for this pathogen and its application to a case study of a Lyme disease patient. The patient had a history of Lyme disease, was treated with antibiotics, and years later developed chronic symptoms including dementia. The patient’s pathology and clinical case description was consistent with Lewy body dementia. B. burgdorferi was identified by PCR in several CNS tissues and by immunofluorescent staining in the spinal cord. These studies offer proof of the principle that persistent infection with the Lyme disease spirochete may have lingering consequences on the CNS.


55. Frontiers in Medicine (Lausanne), Oct 27, 2020
Borrelia miyamotoi Serology in a Clinical Population With Persistent Symptoms and Suspected Tick-Borne Illness https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7652925/
Shannon L. Delaney,1,2,* Lilly A. Murray,1,2 Claire E. Aasen,1 Clair E. Bennett,1,2 Ellen Brown,1,2 and Brian A. Fallon1,2

1 Lyme & Tick-Borne Diseases Research Center, Columbia University Irving Medical Center, New York, NY, United States
2 New York State Psychiatric Institute, Columbia University Irving Medical Center, New York, NY, United States Edited by: Ying Zhang, Zhejiang University, China Reviewed by: Pallab Ghosh, Harvard Medical School, United States; Raymond James Dattwyler, New York Medical College, United States

*Correspondence: Shannon L. Delaney sld2158@cumc.columbia.edu

Abstract: Eighty-two patients seeking consultation for long-term sequalae after suspected tick-borne illness were consecutively tested for Borrelia miyamotoi antibodies using a recombinant glycerophosphodiester phosphodiesterase (GlpQ) enzyme immunoassay. Twenty-one of the 82 patients (26%) tested positive on the GlpQ IgG ELISA. Nearly all of the patients (98%) had no prior B. miyamotoi testing, indicating that clinicians rarely test for this emerging tick-borne pathogen. Compared to patients who solely tested positive for Lyme disease antibodies, patients with B. miyamotoi antibodies presented with significantly more sleepiness and pain. A prospective study is needed to ascertain the relationship between the presence of B. miyamotoi antibodies and persistent symptoms.


54. Antibiotics (Basel). May 25, 2020

Effect of Borrelia burgdorferi Outer Membrane Vesicles on Host Oxidative Stress Response https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7277464/

Keith Wawrzeniak, Gauri Gaur, Eva Sapi, and Alireza G. Senejani*

Department of Biology and Environmental Science, University of New Haven, West Haven, CT 06516, USA; kwawr1@unh.newhaven.edu (K.W.); ggaur2@unh.newhaven.edu (G.G.); ESapi@newhaven.edu (E.S.)

*Correspondence: ASenejani@newhaven.edu

Abstract: Outer membrane vesicles (OMVs) are spherical bodies containing proteins and nucleic acids that are released by Gram-negative bacteria, including Borrelia burgdorferi, the causative agent of Lyme disease. The functional relationship between B. burgdorferi OMVs and host neuron homeostasis is not well understood. The objective of this study was to examine how B. burgdorferi OMVs impact the host cell environment. First, an in vitro model was established by co-culturing human BE2C neuroblastoma cells with B. burgdorferi B31. B. burgdorferi was able to invade BE2C cells within 24 h. Despite internalization, BE2C cell viability and levels of apoptosis remained unchanged, but resulted in dramatically increased production of MCP-1 and MCP-2 cytokines. Elevated secretion of MCP-1 has previously been associated with changes in oxidative stress. BE2C cell mitochondrial superoxides were reduced as early as 30 min after exposure to B. burgdorferi and OMVs. To rule out whether BE2C cell antioxidant response is the cause of decline in superoxides, superoxide dismutase 2 (SOD2) gene expression was assessed. SOD2 expression was reduced upon exposure to B. burgdorferi, suggesting that B. burgdorferi might be responsible for superoxide reduction. These results suggest that B. burgdorferi modulates cell antioxidant defense and immune system reaction in response to the bacterial infection. In summary, these results show that B. burgdorferi OMVs serve to directly counter superoxide production in BE2C neurons, thereby ‘priming’ the host environment to support B. burgdorferi colonization.

53. Brain, Behavior, & Immunity – Health, January 7, 2020 https://www.sciencedirect.com/science/article/pii/S2666354619300158?via%3Dihub
Anti-lysoganglioside and other anti-neuronal autoantibodies in post-treatment Lyme Disease and Erythema Migrans after repeat infection Brian A.Fallon a,b,* Barbara Strobino, a,b, SeanReim c, JulieStoner d, Madeleine W. Cunningham c a Columbia Psychiatry, Columbia University Irving Medical Center, New York, USA b New York State Psychiatric Institute, 1051 Riverside Drive, New York, USA c Department of Microbiology and Immunology, University of Oklahoma Health Sciences Center, Oklahoma City, USA d Department of Biostatistics, University of Oklahoma Health Sciences Center, Oklahoma City, USA

*Corresponding author: Brian A. Fallon, Columbia University, 1051 Riverside Drive, Unit 69, New York, NY, 10032, USA.

Abstract: Background: Molecular mimicry targeting neural tissue has been reported after Borrelia burgdorferi(Bb) infection. Herein, we investigate whether antineuronal autoantibodies are increased and whether antibody-mediated signaling of neuronal cells is elevated in a cohort of symptomatic adults with a history of Lyme Disease (LD). Methods: Participants (n = 179) included 24 with recent Erythema Migrans (EM) without prior LD, 8 with recent EM and prior LD (EM + prior LD), 119 with persistent post-treatment LD symptoms (PTLS), and 28 seronegative endemic controls with no prior LD history. Antineuronal immunoglobulin G (IgG) titers were measured by standard ELISA and compared with mean titers of normal age-matched sera against lysoganglioside, tubulin, and dopamine receptors (D1R and D2R). Antibody-mediated signaling of calcium calmodulin dependent protein kinase II (CaMKII) activity in a human neuronal cell line (SK-N-SH) was identified in serum. Results: EM + prior LD cases had higher antibody titers than controls for anti-lysoganglioside GM1 (p = 0.002), anti-tubulin (p = 0.03), and anti-D1R (p = 0.02), as well as higher expression in the functional antibody-mediated CaMKII Assay (p = 0.03). The EM cases with no prior history showed no significant differences on any measures. The PTLS cases demonstrated significantly higher titers (p = 0.01) than controls on anti-lysoganglioside GM1, but not for the other measures. Conclusion: The finding of elevated anti-neuronal autoantibodies in our small sample of those with a prior history of Lyme disease but not in those without prior Lyme disease, if replicated in a larger sample, suggests an immune priming effect of repeated infection; the CaMKII activation suggests that antineuronal antibodies have functional significance. The elevation of anti-lysoganglioside antibodies among those with PTLS is of particular interest given the established role of anti-ganglioside antibodies in peripheral and central neurologic diseases. Future prospective studies can determine whether these autoantibodies emerge after Bb infection and whether their emergence coincides with persistent neurologic or neuropsychiatric symptoms.

52. Frontiers in Medicine, December 6, 2019 https://www.frontiersin.org/articles/10.3389/fmed.2019.00283/full
The General Symptom Questionnaire-30 (GSQ-30): A Brief Measure of Multi-System Symptom Burden in Lyme Disease
Brian A. Fallon 1,2*, Nevena Zubcevik 3,4, Clair Bennett 1,2, Shreya Doshi 1,2, Alison W. Rebman 5, Ronit Kishon 1,2, James R. Moeller 1,2, Nadlyne R. Octavien 3 and John N. Aucott 5

1 Department of Psychiatry, Lyme and Tick-Borne Diseases Research Center, Columbia University Irving Medical Center, New York, NY, United States
2 Department of Psychiatry, New York State Psychiatric Institute, New York, NY, United States
3 Department of Physical Medicine and Rehabilitation, Dean Center for Tick borne Illness, Harvard Medical School, Spaulding Rehabilitation Hospital, Boston, MA, United States
4 Department of Physical Medicine and Rehabilitation, Massachusetts General Hospital, Boston, MA, United States
5 Division of Rheumatology, Department of Medicine, Lyme Disease Research Center, Johns Hopkins School of Medicine, Baltimore, MD, United States

*Correspondence: Brian A. Fallon, baf1@cumc.columbia.edu

Abstract: Introduction: The multi-system symptoms accompanying acute and post-treatment Lyme disease syndrome pose a challenge for time-limited assessment. The General Symptom Questionnaire (GSQ-30) was developed to fill the need for a brief patient-reported measure of multi-system symptom burden. In this study we assess the psychometric properties and sensitivity to change of the GSQ-30. Materials and Methods: 342 adult participants comprised 4 diagnostic groups: Lyme disease (post-treatment Lyme disease syndrome, n = 124; erythema migrans, n = 94); depression, n = 36; traumatic brain injury, n = 51; healthy, n = 37. Participants were recruited from clinical research facilities in Massachusetts, Maryland, and New York. Validation measures for the GSQ-30 included the Patient Health Questionnaire-4 for depression and anxiety, visual analog scales for fatigue and pain, the Sheehan Disability Scale for functional impairment, and one global health question. To assess sensitivity to change, 53 patients with erythema migrans completed the GSQ-30 before treatment and 6 months after 3 weeks of treatment with doxycycline. Results: The GSQ-30 demonstrated excellent internal consistency (Cronbach α = 0.95). The factor structure reflects four core domains: pain/fatigue, neuropsychiatric, neurologic, and viral-like symptoms. Symptom burden was significantly associated with depression (rs = 0.60), anxiety (rs = 0.55), pain (rs = 0.75), fatigue (rs = 0.77), functional impairment (rs = 0.79), and general health (rs = −0.58). The GSQ-30 detected significant change in symptom burden before and after antibiotic therapy; this change correlated with change in functional impairment. The GSQ-30 total score significantly differed for erythema migrans vs. three other groups (post-treatment Lyme disease syndrome, depression, healthy controls). The GSQ-30 total scores for traumatic brain injury and depression were not significantly different from post-treatment Lyme disease syndrome. Conclusions and Relevance: The GSQ-30 is a valid and reliable instrument to assess symptom burden among patients with acute and post-treatment Lyme disease syndrome and is sensitive in the detection of change after treatment among patients with erythema migrans. The GSQ-30 should prove useful in clinical and research settings to assess multi-system symptom burden and to monitor change over time. The GSQ-30 may also prove useful in future precision medicine studies as a clinical measure to correlate with disease-relevant biomarkers.


51. Meta Gene, Volume 21, September 2019. Online May 2019 https://www.sciencedirect.com/science/article/pii/S2214540019300489
Genetic Variation in the ABCB1 Gene Associated with Post Treatment Lyme Disease Syndrome Status
Joanna Lyon 1, Hyunuk Seung 2

1 Advanced Clinical Pharmacist, University of Maryland School of Pharmacy, United States of America Jlyon@rx.umaryland.edu
2 Department of Pharmacy Practice and Science, University of Maryland School of Pharmacy, United States of America hseung@rx.umaryland.edu

Abstract: Post Treatment Lyme Disease Syndrome (PTLDS) poses a difficult to understand health issue. This multi-centered, randomized control trial studied the possible correlation between ABCB1 (MDR1) gene variants and the incidence of PTLDS in affected patients. Genomic DNA was isolated and analyzed for four ABCB1 gene SNPs (rs1128503, rs1045642, rs2235067, and rs4148740). Significant findings include the association of rs1128503 TC variant with PTLDS status. Additionally, the rs1128503+ rs1045642+ rs2235067 SNP combination increased rs1128503 genotype TC significance to 3.83 times the rs1128503 genotype CC. The TT variant of rs4148740 in conjunction with rs1128503 reduced the odds ratio and appeared to convey a PTLDS protective status to the rs1128503 TC variant.


50. Ticks and Tick-Borne Diseases, 2018 Nov 27 https://www.ncbi.nlm.nih.gov/pubmed/30503356https://www.ncbi.nlm.nih.gov/pubmed/30503356
Regional prevalences of Borrelia burgdorferi, Borrelia bissettiae, and Bartonella henselae in Ixodes affinis, Ixodes pacificus and Ixodes scapularis in the USA.
Maggi RG 1, Toliver M 2, Richardson T 3, Mather T 4, Breitschwerdt EB 5

1 Intracellular Pathogens Research Laboratory, Comparative Medicine Institute, College of Veterinary Medicine, North Carolina State University (NCSU), 1060 William Moore Drive, Raleigh, NC 27607. rgmaggi@ncsu.edu
2 Public Health Pest Management Section, NC Department of Environment and Natural Resources, 10005 Waterford Court, Raleigh, NC 27613. marceetoliver@gmail.com
3 Intracellular Pathogens Research Laboratory, Comparative Medicine Institute, College of Veterinary Medicine, North Carolina State University (NCSU), 1060 William Moore Drive, Raleigh, NC 27607. tgrichar@ncsu.edu
4 Department of Plant Sciences and Entomology, Center for Vecto-Borne Disease, College of the Environment and Life Sciences, University of Rhode Island, 231 Woodward Hall, Kingston, RI 02881. tmather@uri.edu
5 Intracellular Pathogens Research Laboratory, Comparative Medicine Institute, College of Veterinary Medicine, North Carolina State University (NCSU), 1060 William Moore Drive, Raleigh, NC 27607. ebbreits@ncsu.edu

ABSTRACT: The objective of this work was to determine the prevalence of Borrelia and Bartonella species in Ixodes spp. ticks collected from 16 USA states. Genus PCR amplification and sequence analysis of Bartonella and Borrelia 16SsRNA-23SsRNA intergenic regions were performed on DNA extracted from 929 questing adult ticks (671 Ixodes scapularis, 155 Ixodes affinis, and 103 Ixodes pacificus). Overall, 129/929 (13.9%) Ixodes ticks were PCR positive for Borrelia burgdorferi sensu stricto, 48/929 for B. bissettiae whereas 23/929 (2.5%) were PCR positive for a Bartonella henselae. Borrelia bissettiae or B. burgdorferi s.s. and B. henselae co-infections were found in I. affinis from North Carolina at a rate of 4.5%; in a single I. scapularis from Minnesota, but not in I. pacificus. For both bacterial genera, PCR positive rates were highly variable depending on geographic location and tick species, with Ixodes affinis (n = 155) collected from North Carolina, being the tick species with the highest prevalence’s for both Borrelia spp. (63.2%) and B. henselae (10.3%). Based on the results of this and other published studies, improved understanding of the enzootic cycle, transmission dynamics, and vector competence of Ixodes species (especially I. affinis) for transmission of Borrelia spp. and B. henselae should be a public health research priority.


49. Archives of Clinical Neuropsychology, Nov 12, 2018
https://academic.oup.com/acn/advance-article-abstract/doi/10.1093/arclin/acy083/5173742?redirectedFrom=PDF
Neurocognition in Post-Treatment Lyme Disease and Major Depressive Disorder Keilp JG 1,2,3*, Corbera K 1, Gorlyn M 2, Oquendo MA 2,3,4, Mann JJ 2,3

Fallon BA 1,2,3 *Corresponding author at: New York State Psychiatric Institute and Department of Psychiatry, Columbia University College of Physicians and Surgeons, Box 42, NYSPI, 1051 Riverside Drive, New York, NY 10032, USA. Tel.: 1-646-774-7509. E-mail: jgk13@cumc.columbia.edu (J.G. Keilp)

1 Lyme Disease Research Center, Columbia University College of Physicians and Surgeons, New York, NY
2 Department of Psychiatry, Columbia University College of Physicians and Surgeons, New York, NY
3 Department of Molecular Imaging and Neuropathology, New York State Psychiatric Institute, New York, NY
4 Department of Psychiatry, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA

Dr. John Keilp, senior neuropsychologist for the Columbia Lyme & Tick-borne Diseases Research Center’, reports that the cognitive profile of patients with post treatment Lyme disease is meaningfully different from the profile of patients with major depression. This is a neurocognitive biomarker or fingerprint of post-treatment Lyme disease. Although both groups might have fatigue and mental fogginess, the Lyme group more often reports problems with verbal memory and verbal fluency while the depressed (non-Lyme) group more often reports slower processing speed and poor attention. These results highlight the value of neurocognitive testing in helping to tease out the potential causes of cognitive problems in patients with post-treatment Lyme disease.


48. Psychosomatics 59(5) · Sept/Oct 2018 https://www.ncbi.nlm.nih.gov/pubmed/29606281
Depressive Symptoms and Suicidal Ideation Among Symptomatic Patients with a History of Lyme Disease Versus Two Comparison Groups
Shreya Doshi M.A 1*., John G. Keilp Ph.D 2., Barbara Strobino Ph.D. 1, Martin McElhiney Ph.D. 1, Judith Rabkin Ph.D. 1, Brian A. Fallon M.D. 1

1 Division of Clinical Therapeutics, Department of Psychiatry, New York State Psychiatric Institute, New York, NY 2 Division of Molecular Imagining and Neuropathology, Department of Psychiatry, New York State Psychiatric Institute, New York, NY *sd2698@tc.columbia.edu

ABSTRACT: Background: Depression has been reported in 8–45% of patients with posttreatment Lyme symptoms (PTLS), but little is known about suicidal ideation in these patients. Method: Depression and suicidal ideation were assessed using the Beck Depression Inventory (BDI-II). Scores from the PTLS group (n = 81) were compared to those from 2 other groups: HIV+ patients being treated for fatigue (n = 70), and a nonpatient comparison group (NPCG; n = 44). ANOVA and t-tests were used to compare groups; logistic regression was used to identify the strongest correlates of suicidal ideation. Results: Mean BDI-II scores fell in the mildly depressed range for PTLS and HIV+ patients, with both groups having higher depression scores than the NPCG. Suicidal ideation was reported by 19.8% of the PTLS patients and 27.1% of the HIV+ patients, a nonsignificant difference. Among those with mild or no depression, suicidal ideation was uncommon (6.5% PTLS and 11.9% HIV+). Among the patients with moderate-to-severe depression, suicidal ideation was more common (63.2% of 19 PTLS and 50% of 28 HIV+); among these, 2 with PTLS and 1 with HIV+ expressed suicidal intent. Further, 4.5% (n = 2) of the NPCG had suicidal ideation, each had scores in the moderate-to-severe depression range. Higher scores on the cognitive symptoms subscale of the BDI-II predicted greater likelihood of suicidal ideation across patient groups. Conclusion:As expected, suicidal ideation is increased among patients who are depressed. The fact that 1 in 5 patients with PTLS reported suicidal ideation highlights the importance of screening for depression and suicidality to optimize patient care.


47. Healthcare (Basel) 2018 Jun; 6(2): 69. A Community Study of Borrelia burgdorferi Antibodies among Individuals with Prior Lyme Disease in Endemic Areas https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6023339/

Barbara Strobino 1,2,* Katja Steinhagen 3, Wolfgang Meyer 3, Thomas Scheper 3, Sandra Saschenbrecker 3, Wolfgang Schlumberger 3, Winfried Stöcker 3, Andrea Gaito 4 and Brian A. Fallon 1.

1 Department of Psychiatry, Lyme and Tick-Borne Diseases Research Center, Columbia University Irving Medical Center, New York, NY 10032, USA; baf1@cumc.columbia.edu
2 Research Foundation for Mental Hygiene, Inc., New York, NY 10032, USA
3 Institute for Experimental Immunology, Euroimmun, 23560 Lübeck, Germany; k.steinhagen@euroimmun.de (K.S.); w.meyer@euroimmun.de (W.M.); t.scheper@euroimmun.de (T.S.); s.saschenbrecker@euroimmun.de (S.S.); w.schlumberger@euroimmun.de (Wo.S.); w.stoecker@euroimmun.de (Wi.S.)
4 Independent Researcher, Basking Ridge, NJ 07920, USA; adg0339@gmail.com

* Correspondence: barbara.strobino@nyspi.columbia.edu; Tel.: +1-646-774-8052

Abstract: The objective was to examine the prevalence of Borrelia antibodies among symptomatic individuals with recent and past Lyme disease in endemic communities using standard assays and novel assays employing next-generation antigenic substrates. Single- and two-tiered algorithms included different anti-Borrelia ELISAs and immunoblots. Antibody prevalence was examined in sera from 32 individuals with recent erythema migrans (EM), 335 individuals with persistent symptoms following treatment for Lyme disease (PTLS), and 41 community controls without a history of Lyme disease. Among convalescent EM cases, sensitivity was highest using the C6 ELISA (93.8%) compared to other single assays; specificity was 92.7% for the C6 ELISA vs. 85.4–97.6% for other assays. The two-tiered ELISA-EUROLINE IgG immunoblot combinations enhanced case detection substantially compared to the respective ELISA-IgGWestern blot combinations (75.0% vs. 34.4%) despite similar specificity (95.1% vs. 97.6%, respectively). For PTLS cohorts, two-tier ELISA-IgG-blot positivity ranged from 10.1% to 47.4%, depending upon assay combination, time from initial infection, and clinical history. For controls, the two-tier positivity rate was 0–14.6% across assays. A two-tier algorithm of two-ELISA assays yielded a high positivity rate of 87.5% among convalescent EM cases with specificity of 92.7%. For convalescent EM, combinations of the C6 ELISA with a second-tier ELISA or line blot may provide useful alternatives to WB-based testing algorithms.


46. Bio-Protocol. Vol 7, Iss 23, December 05, 2017 DOI: 10.21769/BioProtoc.2643.
https://bio-protocol.org/e2643
Lentiviral Knockdown of Transcription Factor STAT1 in Peromyscus leucopus to Assess Its Role in the Restriction of Tick-borne Flaviviruses
Adaeze O. Izuogu,1 and R. Travis Taylor1,* *travis.taylor@utoledo.edu

1 Department of Medical Microbiology and Immunology, University of Toledo College of Medicine and Life Sciences, Toledo, Ohio, USA

ABSTRACT: Cellular infection with tick-borne flaviviruses (TBFVs) results in activation of the interferon (IFN) signaling pathway and subsequent upregulation of numerous genes termed IFN stimulated genes (ISGs) (Schoggins et al., 2011). Many ISGs function to prevent virus pathogenesis by acting in a broad or specific manner through protein-protein interactions (Duggal and Emerman, 2012). The potency of the IFN signaling response determines the outcome of TBFV infection (Best, 2017; Carletti et al., 2017). Interestingly, data from our lab show that TBFV replication is significantly restricted in cells of the reservoir species Peromyscus leucopus thereby suggesting a potent antiviral response (Izuogu et al., 2017). We assessed the relative contribution of IFN signaling to resistance in P. leucopus by knocking down a major transcription factor in the IFN response pathway. Signal transducer and activator of transcription 1 (STAT1) was specifically targeted in P. leucopus cells by shRNA technology. We further tested the impact of gene knockdown on the ability of cells to respond to IFN and restrict virus replication; the results indicate that when STAT1 expression is altered, P. leucopus cells have a decreased response to IFN stimulation and are significantly more susceptible to TBFV replication.


45. FEBS J. 2017 Nov;284(21):3662-3683. Epub 2017 Sep 30. https://www.ncbi.nlm.nih.gov/pubmed/28892294
Nuclease activity gives an edge to host-defense peptide piscidin 3 over piscidin 1, rendering it more effective against persisters and biofilms.
Libardo MDJ1, Bahar AA2, Ma B3, Fu R4, McCormick LE5, Zhao J6, McCallum SA7, Nussinov R3,8, Ren D2,9,10,11, Angeles-Boza* AM1, Cotten* ML12. *alfredo.angeles-boza@uconn.edu, *mcotten@wm.edu

1 Department of Chemistry, University of Connecticut, Storrs, CT, USA.
2 Department of Biomedical and Chemical Engineering, Syracuse University, NY, USA.
3 Basic Science Program, Leidos Biomedical Research, Inc. Cancer and Inflammation Program, National Cancer Institute, Frederick, MD, USA.
4 National High Magnetic Field Laboratory, Tallahassee, FL, USA.
5 Hamilton College, Department of Chemistry, Clinton, NY, USA.
6 Cancer and Inflammation Program, National Cancer Institute, Frederick, MD, USA.
7 Rennselaer Polytechnic Institute, Center for Biotechnology & Interdisciplinary Studies, Troy, NY, USA.
8 Sackler Institute of Molecular Medicine, Department of Human Genetics and Molecular Medicine, Sackler School of Medicine, Tel Aviv University, Israel.
9 Syracuse Biomaterials Institute, Syracuse University, NY, USA.
10 Department of Civil and Environmental Engineering, Syracuse University, NY, USA. 11 Department of Biology, Syracuse University, NY, USA. 12 Department of Applied Science, College of William and Mary, Williamsburg, VA, USA.

ABSTRACT: Host-defense peptides (HDPs) feature evolution-tested potency against life-threatening pathogens. While piscidin 1 (p1) and piscidin 3 (p3) are homologous and potent fish HDPs, only p1 is strongly membranolytic. Here, we hypothesize that another mechanism imparts p3 strong potency. We demonstrate that the N-termini of both peptides coordinate Cu2+ and p3-Cu cleaves isolated DNA at a rate on par with free Cu2+ but significantly faster than p1-Cu. On planktonic bacteria, p1 is more antimicrobial but only p3 features copper-dependent DNA cleavage. On biofilms and persister cells, p3-Cu is more active than p1-Cu, commensurate with stronger peptide-induced DNA damage. Molecular dynamics and NMR show that more DNA-peptide interactions exist with p3 than p1, and the peptides adopt conformations simultaneously poised for metal- and DNA-binding. These results generate several important conclusions. First, homologous HDPs cannot be assumed to have identical mechanisms since p1 and p3 eradicate bacteria through distinct relative contributions of membrane and DNA-disruptive effects. Second, the nuclease and membrane activities of p1 and p3 show that naturally occurring HDPs can inflict not only physicochemical but also covalent damage. Third, strong nuclease activity is essential for biofilm and persister cell eradication, as shown by p3, the homolog more specific toward bacteria and more expressed in vascularized tissues. Fourth, p3 combines several physicochemical properties (e.g., Amino Terminal Copper and Nickel binding motif; numerous arginines; moderate hydrophobicity) that confer low membranolytic effects, robust copper-scavenging capability, strong interactions with DNA, and fast nuclease activity. This new knowledge could help design novel therapeutics active against hard-to-treat persister cells and biofilms.


44. ACS Chem. Biol., 2017, 12 (5), pp 1170–1182 Publication Date (Web): March 29, 2017 https://pubs.acs.org/doi/abs/10.1021/acschembio.7b00237
Membrane Oxidation in Cell Delivery and Cell Killing Applications
Ting-Yi Wang,†,⊥ M. Daben J. Libardo,‡,⊥ Alfredo M. Angeles-Boza,*,‡ and Jean-Philippe Pellois*,†,§ †Department of Biochemistry and Biophysics, Texas A&M University, College Station, Texas 77843, United States ‡Department of Chemistry, University of Connecticut, Storrs, Connecticut 06269, United States §Department of Chemistry, Texas A&M University, College Station, Texas 77843, United States *E-mail: pellois@tamu.edu., *E-mail: alfredo.angeles-boza@uconn.edu.

ABSTRACT: Cell delivery or cell killing processes often involve the crossing or disruption of cellular membranes. We review how, by modifying the composition and properties of membranes, membrane oxidation can be exploited to enhance the delivery of macromolecular cargoes into live human cells. We also describe how membrane oxidation can be utilized to achieve efficient killing of bacteria by antimicrobial peptides. Finally, we present recent evidence highlighting how membrane oxidation is intimately engaged in natural biological processes such as antigen delivery in dendritic cells and in the killing of bacteria by antimicrobial peptides. Overall, the insights that have been recently gained in this area should facilitate the development of more effective delivery technologies andantimicrobial therapeutic approaches.


43. Biochemistry, 2017, 56 (10), pp 1403–1414 Publication Date (Web): February 22, 2017 https://pubs.acs.org/doi/abs/10.1021/acs.biochem.6b01046
Exploration of the Innate Immune System of Styela clava: Zn2+ Binding Enhances the Antimicrobial Activity of the Tunicate Peptide Clavanin A
Samuel A. Juliano†, Scott Pierce†, James A. deMayo‡, Marcy J. Balunas‡, and Alfredo M. Angeles-Boza*† † Department of Chemistry, University of Connecticut, Storrs, Connecticut 06269-3060, United States ‡ Division of Medicinal Chemistry, Department of Pharmaceutical Sciences, University of Connecticut, Storrs, Connecticut 06269, United States *E-mail: alfredo.angeles-boza@uconn.edu.

ABSTRACT: Tunicates have been used as primitive models for understanding cellmediated and humoral immunity. Clavanin A (ClavA) is one member of a family of antimicrobial peptides produced by the solitary tunicate Styela clava. In this work, we demonstrate that ClavA utilizes Zn2+ ions to potentiate its antimicrobial activity not only by reducing the concentration at which the peptide inhibits the growth of bacteria but also by increasing the rate of killing. Membrane depolarization, β-galactosidase leakage, and potassium leakage assays indicate that ClavA is membrane active, forms small pores, but induces cell death by targeting an intracellular component. ClavA and ClavA-Zn2+ added to Escherichia coli and imaged by confocal microscopy translocate across the cell membrane. E. coli mutants lacking the functional Zn2+ import system are less susceptible to ClavA, suggesting that the synergistic activity between ClavA and Zn2+ has a cytoplasmic target, which is further supported by its nucleolytic activity. Overall, these studies identify a remarkable new mechanism by which zinc contributes to the immune response in the tunicate S. clava.


42. PLoS One, June 26, 2017;12(6)
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5484488/
Interferon Signaling in Peromyscus Leucopus Confers a Potent and Specific Restriction to Vector-borne Flaviviruses
Adaeze O. Izuogu1, Kristin L. McNally2, Stephen E. Harris3, Brian H. Youseff1, John B. Presloid1, Christopher Burlak4, Jason Munshi-South5, Sonja M. Best2, R. Travis Taylor1*

1 Department of Medical Microbiology and Immunology, University of Toledo College of Medicine and Life Sciences, Toledo, Ohio, United States of America,
2 Innate Immunity and Pathogenesis Unit, Laboratory of Virology, Rocky Mountain Laboratories, DIR, NIAID, NIH, Hamilton, Montana, United States of America,
3 The Graduate Center, City University of New York, New York, New York, United States of America,
4 Department of Surgery, University of Minnesota, Minneapolis, Minnesota, United States of America,
5 Louis Calder Center-Biological Field Station, Fordham University, Armonk, New York, United States of America * travis.taylor@utoledo.edu

Abstract: Tick-borne flaviviruses (TBFVs), including Powassan virus and tick-borne encephalitis virus cause encephalitis or hemorrhagic fevers in humans with case-fatality rates ranging from 1-30%. Despite severe disease in humans, TBFV infection of natural rodent hosts has little noticeable effect. Currently, the basis for resistance to disease is not known. We hypothesize that the coevolution of flaviviruses with their respective hosts has shaped the evolution of potent antiviral factors that suppress virus replication and protect the host from lethal infection. In the current study, we compared virus infection between reservoir host cells and related susceptible species. Infection of primary fibroblasts from the white-footed mouse (Peromyscus leucopus, a representative host) with a panel of vector-borne flaviviruses showed up to a 10,000-fold reduction in virus titer compared to control Mus musculus cells. Replication of vesicular stomatitis virus was equivalent in P. leucopus and M. musculus cells suggesting that restriction was flavivirus-specific. Step-wise comparison of the virus infection cycle revealed a significant block to viral RNA replication, but not virus entry, in P. leucopus cells. To understand the role of the type I interferon (IFN) response in virus restriction, we knocked down signal transducer and activator of transcription 1 (STAT1) or the type I IFN receptor (IFNAR1) by RNA interference. Loss of IFNAR1 or STAT1 significantly relieved the block in virus replication in P. leucopus cells. The major IFN antagonist encoded by TBFV, nonstructural protein 5, was functional in P. leucopus cells, thus ruling out ineffective viral antagonism of the host IFN response. Collectively, this work demonstrates that the IFN response of P. leucopus imparts a strong and virus-specific barrier to flavivirus replication. Future identification of the IFN-stimulated genes responsible for virus restriction specifically in P. leucopus will yield mechanistic insight into efficient control of virus replication and may inform the development of antiviral therapeutics.


41. Antibiotics, March 22, 2017
http://www.mdpi.com/2079-6382/6/1/10/pdf
Activity of Sulfa Drugs and Their Combinations against Stationary Phase B. burgdorferi In Vitro Jie Feng, Shuo Zhang, Wanliang Shi and Ying Zhang * Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD 21205, USA *Author to whom correspondence should be addressed. yzhang@jhsph.edu

Abstract: Lyme disease is a most common vector-borne disease in the US. Although the majority of Lyme patients can be cured with the standard two- to four-week antibiotic treatment, at least 10%–20% of patients continue to suffer from prolonged post-treatment Lyme disease syndrome (PTLDS). While the cause for this is unclear, one possibility is that persisting organisms are not killed by current Lyme antibiotics. In our previous studies, we screened an FDA drug library and an NCI compound library on B. burgdorferi and found some drug hits including sulfa drugs as having good activity against B. burgdorferi stationary phase cells. In this study, we evaluated the relative activity of three commonly used sulfa drugs, sulfamethoxazole (Smx), dapsone (Dps), sulfachlorpyridazine (Scp), and also trimethoprim (Tmp), and assessed their combinations with the commonly prescribed Lyme antibiotics for activities against B. burgdorferi stationary phase cells. Using the same molarity concentration, dapsone, sulfachlorpyridazine and trimethoprim showed very similar activity against stationary phase B. burgdorferi enriched in persisters; however, sulfamethoxazole was the least active drug among the three sulfa drugs tested. Interestingly, contrary to other bacterial systems, Tmp did not show synergy in drug combinations with the three sulfa drugs at their clinically relevant serum concentrations against B. burgdorferi. We found that sulfa drugs combined with other antibiotics were more active than their respective single drugs and that four-drug combinations were more active than three-drug combinations. Four-drug combinations dapsone + minocycline + cefuroxime + azithromycin and dapsone + minocycline + cefuroxime + rifampin showed the best activity against stationary phase B. burgdorferi in these sulfa drug combinations. However, these four-sulfa-drug–containing combinations still had considerably less activity against B. burgdorferi stationary phase cells than the Daptomycin + cefuroxime + doxycycline used as a positive control which completely eradicated B. burgdorferi stationary phase cells. Future studies are needed to evaluate and optimize the sulfa drug combinations in vitro and also in animal models.


40. Frontiers in Microbiology, October 19, 2016 http://journal.frontiersin.org/article/10.3389/fmicb.2016.01744/pdf
Ceftriaxone Pulse Dosing Fails to Eradicate Biofilm-like Microcolony B. burgdorferi Persisters Which Are Sterilized by Daptomycin/Doxycycline/Cefuroxime Drug Combination without Pulse Dosing
Jie Feng1, Shuo Zhang1, Wanliang Shi1, Ying Zhang1*

1Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, USA *Corresponding author: Ying Zhang, MD, PhD Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD 21205, USA yzhang@jhsph.edu

Abstract: Although the majority of Lyme disease patients can be cured, at least 10-20% of the patients continue to suffer from persisting symptoms such as fatigue, muscular and joint pain, and neurologic impairment after standard 2-4 week antibiotic treatment. While the causes for this post-treatment Lyme disease symptoms are unclear, one possibility is due to B. burgdorferi persisters that are not effectively killed by current antibiotics such as doxycycline or amoxicillin used to treat Lyme disease. A previous study showed that four rounds of ceftriaxone pulse dosing treatment eradicated B. burgdorferi persisters in vitro using a relatively young late log phase culture (5 day old). In this study, we investigated if ceftriaxone pulse dosing could also eradicate B. burgdorferi persisters in older stationary phase cultures (10 day old) enriched with more resistant microcolony form of persisters. We found that ceftriaxone pulse dosing could only eradicate planktonic log phase B. burgdorferi spirochetal forms and round body forms but not more resistant aggregated biofilm-like microcolony persisters enriched in stationary phase cultures. Moreover, we found that not all drugs are suitable for pulse dosing, with bactericidal drugs ceftriaxone and cefuroxime being more appropriate for pulse dosing than bacteriostatic drug doxycycline and persister drug daptomycin. We also showed that drug combination pulse dosing treatment was more effective than single drug pulse dosing. Importantly, we demonstrate that pulse dosing treatment impaired the activity of the persister drug daptomycin and its drug combination against B. burgdorferi persisters and that the most effective way to kill the more resistant biofilm-like microcolonies is the daptomycin/doxycycline/ceftriaxone triple drug combination without pulse dosing. Our findings indicate pulse dosing may not always work as a general principle but rather depends on the specific drugs used, with cidal drugs being more appropriate for pulse dosing than static or persister drugs, and that drug combination approach with persister drugs is more effective at killing the more resistant microcolony form of persisters than pulse dosing. These observations may have implications for more effective treatment of Lyme disease. Future studies are required to validate these findings in animal models of B. burgdorferi persistence.


39. Frontiers in Microbiology, May 23, 2016 http://journal.frontiersin.org/article/10.3389/fmicb.2016.00743/full
A Drug Combination Screen Identifies Drugs Active against Amoxicillin-Induced Round Bodies of In Vitro Borrelia burgdorferi Persisters from an FDA Drug Library Ying Zhang,
Jie Feng, Wanliang Shi, Shuo Zhang, David Sullivan, Paul G. Auwaerter, Johns Hopkins

Abstract: Although currently recommended antibiotics for Lyme disease such as doxycycline or amoxicillin cure the majority of the patients, about 10–20% of patients treated for Lyme disease may experience lingering symptoms including fatigue, pain, or joint and muscle aches. Under experimental stress conditions such as starvation or antibiotic exposure, Borrelia burgdorferi can develop round body forms, which are a type of persister bacteria that appear resistant in vitro to customary first-line antibiotics for Lyme disease. To identify more effective drugs with activity against the round body form of B. burgdorferi, we established a round body persister model induced by exposure to amoxicillin (50 μg/ml) and then screened the Food and Drug Administration drug library consisting of 1581 drug compounds and also 22 drug combinations using the SYBR Green I/propidium iodide viability assay. We identified 23 drug candidates that have higher activity against the round bodies of B. burgdorferi than either amoxicillin or doxycycline. Eleven individual drugs scored better than metronidazole and tinidazole which have been previously described to be active against round bodies. In this amoxicillin-induced round body model, some drug candidates such as daptomycin and clofazimine also displayed enhanced activity which was similar to a previous screen against stationary phase B. burgdorferi persisters not exposure to amoxicillin. Additional candidate drugs active against round bodies identified include artemisinin, ciprofloxacin, nifuroxime, fosfomycin, chlortetracycline, sulfacetamide, sulfamethoxypyridazine and sulfathiozole. Two triple drug combinations had the highest activity against amoxicillin-induced round bodies and stationary phase B. burgdorferi persisters: artemisinin/cefoperazone/doxycycline and sulfachlorpyridazine/daptomycin/doxycycline. These findings confirm and extend previous findings that certain drug combinations have superior activity against B. burgdorferi persisters in vitro, even when pre-treated with amoxicillin. These findings may have implications for improved treatment of Lyme disease.


38. Park Science (NPS, Department of Interior) March 2016 http://www.nature.nps.gov/ParkScience/Archive/PDF/Article_PDFs/ParkScience32(1)Summer2015_36-41_Ford_et_al_3819.pdf
Tick surveillance and disease prevention on the Appalachian Trail (Also published in Appalachian Trail Journeys, The Magazine of the Appalachian Trail Conservancy, May/June 2014)

Karl Ford, Robyn Nadolny, Ellen Stromdahl, and Graham Hickling Abstract: The Appalachian National Scenic Trail (AT) runs 3,520 km (2,187 mi) from northern Georgia to northern Maine, traversing 14 states where Lyme disease and other tickborne diseases are endemic or emerging. Approximately 2–3 million visitors hike the AT annually, including through-hikers who spend five to six months on the trail in spring through early fall, when common tick species are active. Disease vector tick surveillance was conducted from April through August 2013 at 42 randomly selected AT shelter areas along a south-to-north transect covering the full length of the AT. Tick abundance at shelters and tenting areas was compared with tick abundance on the AT itself, and the collected ticks were tested for common bacterial pathogens. Human-biting tick species collected comprised Ixodes scapularis, Amblyomma americanum, Amblyomma maculatum, and Dermacentor variabilis. Human pathogens Borrelia burgdorferi and Rickettsia montanensis were detected in tested ticks. Tick abundance on the trail was low overall (2.8 ticks per 1,000 m2 sampled), but exceeded tick abundance in shelters and tenting areas by 14.5 times. No ticks were collected south of Virginia or north of Massachusetts, or above 829 m (2,720 ft) in elevation, which suggests that season and elevation are significant determinants of the risk of hiker exposure to questing ticks on the AT. Such information should be included in future health messaging to hikers along with preventive measures. Management issues are discussed.


37. FEMS Microbiology Letters Advance Access, July 24, 2015 http://femsle.oxfordjournals.org/content/early/2015/07/23/femsle.fnv120
Biofilm formation by 1 Borrelia sensu lato Arun Timmaraju1,2,†, Priyanka A.S. Theophilus1,†, Kunthavai Balasubramanian1,3, Shafiq Shakih1, David F. Leucke1 and Eva Sapi1,*

1 Lyme disease research group, Department of Biology and Environmental Science, University of New Haven, West Haven, CT, USA
2 Present address: Interpace Diagnostics, New Haven, CT, 06519 3 Present address: Department of Hematology, Yale School of Medicine, New Haven, CT, 06520 † Contributed equally * Correspondence: Eva Sapi Ph.D., Department of Biology and Environmental Sciences, University of New Haven, 1211 Campbell Avenue, Charger Plaza LL16. West Haven, CT, 06516, USA. esapi@newhaven.edu

Abstract: Bacterial biofilms are microbial communities held together by an extracellular polymeric substance matrix predominantly composed of polysaccharides, proteins and nucleic acids. We had previously shown that Borrelia burgdorferi sensu stricto, the causative organism of Lyme disease in the United States is capable of forming biofilms in vitro. Here, we investigated biofilm formation by Borrelia afzelii and Borrelia garinii, which cause Lyme disease in Europe. Using various histochemistry and microscopy techniques, we show that Borrelia afzelii and Borrelia garinii form biofilms, which resemble biofilms formed by Borrelia burgdorferi sensu stricto. High-resolution atomic force microscopy revealed similarities in the ultra-structural organization of the biofilms form by three Borrelia species. Histochemical experiments revealed a heterogeneous organization of exopolysaccharides among the three Borrelia species. These results suggest that biofilm formation might be a common trait of Borrelia genera physiology.


36. Clinical Infectious Diseases Advance Access, September 2, 2014 http://www.ncbi.nlm.nih.gov/pubmed/25182244
A comparison of Lyme disease serologic test results from four laboratories in patients with persistent symptoms after antibiotic treatment Brian A. Fallon1, Martina Pavlicova2, Samantha W. Coffino3, Carl Brenner4

1 Department of Psychiatry, Columbia University, New York NY
2 Department of Biostatistics, Mailman School of Public Health, Columbia University, New York, NY
3 Department of Neurology, Columbia University, New York, NY 4 Lamont-Doherty Earth Observatory of Columbia University, Palisades, NY Summary: In patients with post-treatment Lyme syndrome, rates of positive serologic test results were generally similar among a university laboratory, a commercial laboratory, and two Lyme specialty laboratories, although interlaboratory variability was high and the IgM Western Blot performed poorly.

Abstract: Background – As the incidence of Lyme disease (LD) has increased, a number of “Lyme specialty laboratories” have emerged, claiming singular expertise in LD testing. We investigated the degree of interlaboratory variability of several LD serologic tests—whole cell sonicate (WCS) enzyme-linked immunosorbent assay (ELISA), IgM and IgG Western blots (WB), and an ELISA based on the conserved sixth region of VlsE (C6)—performed at one university laboratory, one commercial laboratory and two laboratories that specialize in LD testing.


35. Veterinary Sciences, 2014
www.mdpi.com/2306-7381/1/1/5
Filarial Nematode Infection in Ixodes scapularis Ticks Collected from Southern Connecticut Pabbati Namrata, Jamie M. Miller, Madari Shilpa, Patlolla Raghavender Reddy, Cheryl Bandoski, Michael J. Rossi and Eva Sapi* Department of Biology and Environmental Science, University of New Haven, West Haven, CT 06516, USA

*Author to whom correspondence should be addressed.

Abstract: It was recently demonstrated that the lone star tick Amblyomma americanum could harbor filarial nematodes within the genus Acanthocheilonema. In this study, Ixodes scapularis (deer) ticks collected from Southern Connecticut were evaluated for their potential to harbor filarial nematodes. Non-engorged nymphal and adult stage Ixodes scapularis ticks were collected in Southern Connecticut using the standard drag method. In situ hybridization with filarial nematode specific sequences demonstrated the presence of filarial nematodes in Ixodes ticks. Filarial nematode specific DNA sequences were amplified and confirmed by direct sequencing in Ixodes nymphal and adult ticks using either general filarial nematode or Onchocercidae family specific PCR primers. Phylogenetic analysis of the 12S rDNA gene sequence indicated that the filarial nematode infecting Ixodes scapularis ticks is most closely related to the species found in Amblyomma americanum ticks and belongs to the genus of Acanthocheilonema. Our data also demonstrated that infection rate of these filarial nematode in Ixodes ticks is relatively high (about 22% and 30% in nymphal and adult Ixodes ticks, respectively). In summary, the results from our studies demonstrated that filarial nematode infection was found in Ixodes ticks similar to what has been found in Amblyomma americanum ticks. Vet. Sci. 2014, 1, 5-15; doi:10.3390/vetsci1010005


34. Psychosomatics 2013
http://www.psychosomaticsjournal.com/article/S0033-3182%2813%2900078-9/fulltext
Correlates of Perceived Health-Related Quality of Life in Post-treatment Lyme Encephalopathy Avinash M.Chandra , B.A.-1, John G. Keilp, Ph.D.-2, Brian A. Fallon*, M.D.-2

1 Department of Psychiatry, Columbia University, New York, NY.
2 Division of Clinical Therapeutics of the New York State Psychiatric Institute, New York, NY *Corresponding Author: Brian A. Fallon, M.D baf1@columbia.edu

ABSTRACT: Background – Marked functional impairment has been reported by patients with post-treatment Lyme disease syndrome (PTLDS). Objective: We sought to identify but the clinical features that contribute most strongly to the impaired health status associated with PTLDS. Methods: Enrolled patients had a well-documented history of Lyme disease, prior treatment with at least 3 weeks with intravenous ceftriaxone, a positive IgG Western blot, and objective problems with memory. An index score to capture aggregate cognitive functioning, Short-Form 36 physical and mental component summer scores, and scores on other clinical and demographic measures were examined. Multiple linear regressions were performed to determine significant predictors of perceptions of impaired life functioning as delineated by theShort-Form36.Results: Fatigue was the most important contributor to perceived impairments in overall physical functioning, and fatigue and depression significantly predicted perceived impairments in overall mental functioning. Conclusions: Because fatigue and depression contribute prominently to reports of impaired physical functioning and mental functioning among patients with PTLDS, clinicians should assess patients for these symptoms and consider targeting these symptoms in the selection of treatment interventions. Future controlled studies should examine the effectiveness of such agents for patients with PTLDS.


33. International Journal of Medical Sciences 2013 http://www.medsci.org/v10p0915.htm
Lyme Borreliosis in Human Patients in Florida and Georgia, USA
Kerry L. Clark-1, Brian Leydet-1,2, Shirley Hartman-3 1-University of North Florida, 2-Louisiana State University, 3-Mandarin Wellness Center, Florida Corresponding author: Kerry L. Clark, M.P.H., Ph.D., Department of Public Health, University of North Florida, 1 UNF Drive, Jacksonville, FL 32224. Phone: (904) 620-1427. Fax: (904) 620-2848. E-mail: kclark@unf.edu.

ABSTRACT: The aim of this study was to determine the cause of illness in several human patients residing in Florida and Georgia, USA, with suspected Lyme disease based upon EM-like skin lesions and/or symptoms consistent with early localized or late disseminated Lyme borreliosis. Using polymerase chain reaction (PCR) assays developed specifically for Lyme group Borrelia spp., followed by DNA sequencing for confirmation, we identified Borrelia burgdorferi sensu lato DNA in samples of blood and skin and also in lone star ticks (Amblyomma americanum) removed from several patients who either live in or were exposed to ticks in Florida or Georgia. This is the first report to present combined PCR and DNA sequence evidence of infection with Lyme Borrelia spp. in human patients in the southern U.S., and to demonstrate that several B. burgdorferi sensu lato species may be associated with Lyme disease-like signs and symptoms in southern states. Based on the findings of this study, we suggest that human Lyme borreliosis occurs in Florida and Georgia, and that some cases of Lyme-like illness referred to as southern tick associated rash illness (STARI) in the southern U.S. may be attributable to previously undetected B. burgdorferi sensu lato infections.


32. Northeastern Naturalist 2013; 20(1):197–204.
http://www.bioone.org/doi/abs/10.1656/045.020.0116
Distribution of Ticks & Prevalence of Borrelia burgdorferi in the Upper Connecticut River Valley of Vermont
Abigail C. Serra-1, Paul S. Warden-2, Colin R. Fricker-2, and Alan R. Giese-1,*

1-Lyndon State College VT, 2-Analytical Services, Inc. VT
Corresponding author – giese@lyndonstate. edu.

ABSTRACT: Ixodes scapularis (Black-legged Tick) has expanded its range in recent decades. To establish baseline data on the abundance of the Black-legged Tick and Borrelia burgdorferi (the causative agent of Lyme disease) at the edge of a putative range expansion, we collected 1398 ticks from five locations along the Connecticut River in Vermont. Collection locations were approximately evenly distributed between the villages of Ascutney and Guildhall. Relative abundance and distribution by species varied across sites. Black-legged Ticks dominated our collections (n = 1348, 96%), followed by Haemaphysalis leporispalustris (Rabbit Tick; n = 45, 3%), and Dermacentor variabilis (American Dog Tick; n = 5, <1%). Black-legged Tick abundance ranged from 6198 ticks per survey hectare (all life stages combined) at the Thetford site to zero at the Guildhall site. There was little to no overlap of tick species across sites. Phenology of Black-legged Ticks matched published information from other regions of the northeastern USA. Prevalence of B. burgdorferi in adult Black-legged Ticks was 8.9% (n = 112)


31. J Neuropsychiatry Clin Neurosci. 2013 Feb 27. doi: 10.1176/appi.neuropsych.12090223. http://www.ncbi.nlm.nih.gov/pubmed/23446551

Post-Treatment Lyme Syndrome and Central Sensitization.

Batheja S, Nields JA, Landa A, Fallon BA*. Department of Psychiatry, Columbia University, and The New York State Psychiatric Institute.
*Send correspondence to Dr. Fallon; e-mail: baf1@columbia.edu

ABSTRACT: Central sensitization is a process that links a variety of chronic pain disorders that are characterized by hypersensitivity to noxious stimuli and pain in response to non-noxious stimuli. Among these disorders, treatments that act centrally may have greater efficacy than treatments acting peripherally. Because many individuals with post-treatment Lyme syndrome (PTLS) have a similar symptom cluster, central sensitization may be a process mediating or exacerbating their sensory processing. This article reviews central sensitization, reports new data on sensory hyperarousal in PTLS, explores the potential role of central sensitization in symptom chronicity, and suggests new directions for neurophysiologic and treatment research.


30. The Open Neurology Journal 2012: 6, (Suppl 1-M2) 79-87 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3474942/
A Reappraisal of the U.S. Clinical Trials of Post-Treatment Lyme Disease Syndrome Brian A. Fallon*,1, Eva Petkova2, John G. Keilp3 and Carolyn B. Britton4

1 Columbia University, Dept. of Psychiatry, Division of Clinical Therapeutics, USA
2 New York University, Dept. of Child and Adolescent Psychiatry, Division of Biostatistics, USA
3 Columbia University, Dept. of Psychiatry, Division of Neuroscience, USA
4 Columbia University, Dept. of Neurology, USA *Send correspondence to Dr. Fallon; e-mail: baf1@columbia.edu

ABSTRACT: Four federally funded randomized placebo-controlled treatment trials of post-treatment Lyme syndrome in the United States have been conducted. Most international treatment guidelines summarize these trials as having shown no acute or sustained benefit to repeated antibiotic therapy. The goal of this paper is to determine whether this summary conclusion is supported by the evidence. Methods: The methods and results of the 4 U.S. treatment trials are described and their critiques evaluated. Results: 2 of the 4 U.S. treatment trials demonstrated efficacy of IV ceftriaxone on primary and/or secondary outcome measures. Conclusions: Future treatment guidelines should clarify that efficacy of IV ceftriaxone for post-treatment Lyme fatigue was demonstrated in one RCT and supported by a second RCT, but that its use was not recommended primarily due to adverse events stemming from the IV route of treatment. While repeated IV antibiotic therapy can be effective, safer modes of delivery are needed.


29. PLoS ONE 7(10) 2012: e48277. doi:10.1371/journal.pone.0048277 http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0048277
Characterization of Biofilm Formation by Borrelia burgdorferi In Vitro

Eva Sapi*, Scott L. Bastian, Cedric M. Mpoy, Shernea Scott, Amy Rattelle, Namrata Pabbati, Akhila Poruri, Divya Burugu, Priyanka A. S. Theophilus, Truc V. Pham, Akshita Datar, Navroop K. Dhaliwal, Alan MacDonald, Michael J. Rossi, David F. Luecke (Lyme Disease Research Group, Department of Biology and Environmental Sciences, University of New Haven, West Haven, Connecticut, United States of America); Saion K. Sinha (Department of Physics, University of New Haven, West Haven, Connecticut, United States of America).

*Corresponding Author Email: Eva Sapi, esapi@newhaven.edu

ABSTRACT: Borrelia burgdorferi, the causative agent of Lyme disease, has long been known to be capable of forming aggregates and colonies. It was recently demonstrated that Borrelia burgdorferi aggregate formation dramatically changes the in vitro response to hostile environments by this pathogen. In this study, we investigated the hypothesis that these aggregates are indeed biofilms, structures whose resistance to unfavorable conditions are well documented. We studied Borrelia burgdorferi for several known hallmark features of biofilm, including structural rearrangements in the aggregates, variations in development on various substrate matrices and secretion of a protective extracellular polymeric substance (EPS) matrix using several modes of microscopic, cell and molecular biology techniques. The atomic force microscopic results provided evidence that multilevel rearrangements take place at different stages of aggregate development, producing a complex, continuously rearranging structure. Our results also demonstrated that Borrelia burgdorferi is capable of developing aggregates on different abiotic and biotic substrates, and is also capable of forming floating aggregates. Analyzing the extracellular substance of the aggregates for potential exopolysaccharides revealed the existence of both sulfated and non-sulfated/carboxylated substrates, predominately composed of an alginate with calcium and extracellular DNA present. In summary, we have found substantial evidence that Borrelia burgdorferi is capable of forming biofilm in vitro. Biofilm formation by Borrelia species might play an important role in their survival in diverse environmental conditions by providing refuge to individual cells.


28. PLOS One 2012
http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0033280
Genome Stability of Lyme Disease Spirochetes: Comparative Genomics of Borrelia burgdorferi Plasmids

Sherwood R. Casjens1*, Emmanuel F. Mongodin2, Wei-Gang Qiu3, Benjamin J. Luft4, Steven E. Schutzer5, Eddie B. Gilcrease1, Wai Mun Huang1, Marija Vujadinovic1, John K. Aron1, Levy C. Vargas3, Sam Freeman3, Diana Radune6, Janice F. Weidman6, George I. Dimitrov6, Hoda M. Khouri6, Julia E. Sosa6, Rebecca A. Halpin6, John J. Dunn7, Claire M. Fraser2

1-University of Utah School of Medicine,
2-University of Maryland School of Medicine,
3-Hunter College of the City University of New York,
4-Stony Brook University, NY,
5- New Jersey Medical School,
6-J. Craig Venter Institute, MD,
7-Brookhaven National Laboratory, NY

*Corresponding Author Email: sherwood.casjens@path.utah.edu

ABSTRACT: Lyme disease is the most common tick-borne human illness in North America. In order to understand the molecular pathogenesis, natural diversity, population structure and epizootic spread of the North American Lyme agent, Borrelia burgdorferi sensu stricto, a much better understanding of the natural diversity of its genome will be required. Towards this end we present a comparative analysis of the nucleotide sequences of the numerous plasmids of B. burgdorferi isolates B31, N40, JD1 and 297. These strains were chosen because they include the three most commonly studied laboratory strains, and because they represent different major genetic lineages and so are informative regarding the genetic diversity and evolution of this organism. A unique feature of Borrelia genomes is that they carry a large number of linear and circular plasmids, and this work shows that strains N40, JD1, 297 and B31 carry related but non-identical sets of 16, 20, 19 and 21 plasmids, respectively, that comprise 33–40% of their genomes. We deduce that there are at least 28 plasmid compatibility types among the four strains. The B. burgdorferi ~900 Kbp linear chromosomes are evolutionarily exceptionally stable, except for a short ≤20 Kbp plasmid-like section at the right end. A few of the plasmids, including the linear lp54 and circular cp26, are also very stable. We show here that the other plasmids, especially the linear ones, are considerably more variable. Nearly all of the linear plasmids have undergone one or more substantial inter-plasmid rearrangements since their last common ancestor. In spite of these rearrangements and differences in plasmid contents, the overall gene complement of the different isolates has remained relatively constant.


27. Journal of Bacteriology 2011 http://jb.asm.org/content/193/4/1018.long Whole-Genome Sequences of Thirteen Isolates of Borrelia burgdorferi Steven E. Schutzer1,*, Claire M. Fraser-Liggett2, Sherwood R. Casjens3,*, Wei-Gang Qiu4, John J. Dunn5, Emmanuel F. Mongodin2, and Benjamin J. Luft6 1-University of Medicine and Dentistry of New Jersey—New Jersey Medical School, 2-Institute for Genome Sciences, University of Maryland, School of Medicine, 3-University of Utah Medical School, 4- Hunter College of the City University of New York, 5-Brookhaven National Laboratory, Upton, New York 6-Stony Brook University *Corresponding author. Mailing address for Steven E. Schutzer: Department of Medicine, University of Medicine and Dentistry of New Jersey—New Jersey Medical School, Newark, NJ 07103. E-mail: schutzer@umdnj.edu. Mailing address for Sherwood R. Casjens: Department of Pathology, University of Utah Medical School, Room 2200 EEJMRB, 15 North Medical Dr. East, Salt Lake City, UT 84112. E-mail: sherwood.casjens@path.utah.edu ABSTRACT: Borrelia burgdorferi is a causative agent of Lyme disease in North America and Eurasia. The first complete genome sequence of B. burgdorferi strain 31, available for more than a decade, has assisted research on the pathogenesis of Lyme disease. Because a single genome sequence is not sufficient to understand the relationship between genotypic and geographic variation and disease phenotype, we determined the whole-genome sequences of 13 additional B. burgdorferi isolates that span the range of natural variation. These sequences should allow improved understanding of pathogenesis and provide a foundation for novel detection, diagnosis, and prevention strategies.


26. PLoS ONE 6(2): e17287. doi:10.1371/journal.pone.0017287 2011 http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0017287
Distinct Cerebrospinal Fluid Proteomes Differentiate Post-Treatment Lyme Disease from Chronic Fatigue Syndrome

Steven E. Schutzer-1#*, Thomas E. Angel-4#, Tao Liu-4#, Athena A. Schepmoes-4, Therese R. Clauss-4, Joshua N. Adkins-4, David G. Camp II-4, Bart K. Holland-3, Jonas Bergquist-5, Patricia K. Coyle-6, Richard D. Smith-4, Brian A. Fallon-7, Benjamin H. Natelson-2,8

1-Department of Medicine, University of Medicine and Dentistry of New Jersey-New Jersey Medical School, Newark, New Jersey, United States of America,
2 Department of Neurology, University of Medicine and Dentistry of New Jersey-New Jersey Medical School, Newark, New Jersey, United States of America,
3 Division of Biostatistics and Epidemiology, University of Medicine and Dentistry of New Jersey-New Jersey Medical School, Newark, New Jersey, United States of America,
4 Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, United States of America,
5 Department of Physical and Analytical Chemistry, Uppsala University, Uppsala, Sweden,
6 Department of Neurology, State University of New York-Stony Brook, Stony Brook, New York, United States of America,
7 Department of Psychiatry, Columbia University Medical Center, New York, New York, United States of America, 8 Department of Pain Medicine and Palliative Care and Beth Israel Medical Center, Albert Einstein School of Medicine, Bronx, New York, United States of America

*Corresponding Author Email: schutzer@umdnj.edu #These authors contributed equally to this work

ABSTRACT: Neurologic Post Treatment Lyme disease (nPTLS) and Chronic Fatigue (CFS) are syndromes of unknown etiology. They share features of fatigue and cognitive dysfunction, making it difficult to differentiate them. Unresolved is whether nPTLS is a subset of CFS. Pooled cerebrospinal fluid (CSF) samples from nPTLS patients, CFS patients, and healthy volunteers were comprehensively analyzed using high-resolution mass spectrometry (MS), coupled with immunoaffinity depletion methods to reduce protein-masking by abundant proteins. Individual patient and healthy control CSF samples were analyzed directly employing a MS-based label-free quantitative proteomics approach. We found that both groups, and individuals within the groups, could be distinguished from each other and normals based on their specific CSF proteins (p<0.01). CFS (n = 43) had 2,783 non-redundant proteins, nPTLS (n = 25) contained 2,768 proteins, and healthy normals had 2,630 proteins. Preliminary pathway analysis demonstrated that the data could be useful for hypothesis generation on the pathogenetic mechanisms underlying these two related syndromes. nPTLS and CFS have distinguishing CSF protein complements. Each condition has a number of CSF proteins that can be useful in providing candidates for future validation studies and insights on the respective mechanisms of pathogenesis. Distinguishing nPTLS and CFS permits more focused study of each condition, and can lead to novel diagnostics and therapeutic interventions.


25. Genetics: Published Articles Ahead of Print, published on September 2, 2011 as 0.1534/genetics.111.130773 Copyright 2011 http://www.genetics.org/content/189/3/951.long
Pervasive Recombination and Sympatric Genome Diversification Driven by Frequency-Dependent Selection in Borrelia burgdorferi, the Lyme disease Bacterium
James Haven*,1, Levy C. Vargas†, Emmanuel F. Mongodin‡, Vincent Xue§, Yozen Hernandez†, Pedro Pagan†, Claire M. Fraser-Liggett‡, Steven E. Schutzer**, Benjamin J. Luft††, Sherwood R. Casjens‡‡, and Wei-Gang Qiu†*, §§, 2 *Department of Biology, The Graduate Center, City University of New York, New York, New York 10016 †Department of Biological Sciences and The Center for Gene Structure and Function and §Department of Computer Science, Hunter College, City University of New York, New York, New York 10065 ‡Institute for Genome Sciences, University of Maryland BioPark, Baltimore, Maryland 21201 **Department of Medicine, University of Medicine and Dentistry of New Jersey–New Jersey Medical School, Newark, New Jersey 07103 ††Department of Medicine, Health Science Center, Stony Brook University, Stony Brook, New York 11794 ‡‡Department of Pathology, Division of Molecular Cell Biology and Immunology, University of Utah School of Medicine, Salt Lake City, Utah 84112 §§National Evolutionary Synthesis Center, Durham, North Carolina 27705 1 Present address: Odum School of Ecology, University of Georgia, Athens, GA 30602. 2 Corresponding author: Department of Biological Sciences, Hunter College of the City University of New York, 695 Park Ave., New York, NY 10065. E-mail: weigang@genectr.hunter.cuny.edu

ABSTRACT: How genomic diversity within bacterial populations originates and is maintained in thepresence of frequent recombination is a central problem in understanding bacterial evolution. Natural populations of Borrelia burgdorferi, the bacterial agent of Lyme disease, consist of diverse genomic groups co-infecting single individual vertebrate hosts and tick vectors. To understand mechanisms of sympatric genome differentiation in B. burgdorferi, we sequenced and compared 23 genomes representing major genomic groups in North America and Europe. Linkage analysis of over 13,500 single nucleotide polymorphisms revealed pervasive horizontal DNA exchanges. Although three times more frequent than point mutation, recombination is localized and weakly affects genome-wide linkage disequilibrium. We show by computer simulations that, while enhancing population fitness, recombination constrains neutral and adaptive divergence among sympatric genomes through periodic selective sweeps. In contrast, simulations of frequency-dependent selection with recombination produced the observed pattern of a large number of sympatric genomic groups associated with major sequence variations at the selected locus. We conclude that negative frequency-dependent selection targeting a small number of surface-antigen loci (ospC in particular) sufficiently explains the maintenance of sympatric genome diversity in B. burgdorferi without adaptive divergence. In fact, pervasive recombination makes it unlikely for local B. burgdorferi genomic groups to achieve host specialization. B. burgdorferi genomic groups in northeastern United States are thus best viewed as constituting a single bacterial species, whose generalist nature is a key to its rapid spread and human virulence.


24. Journal of Medical Entomology 47(1):89-94. 2010 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2837073/
Extraction of Total Nucleic Acids from Ticks for the Detection of Bacterial & Viral Pathogens
Chris D. Crowder-1, Megan A. Rounds-1, Curtis A. Phillipson-1, John M. Picuri-1, Heather E. Matthews-1, Justina Halverson-1, Steven E. Schutzer-2, David J. Ecker-1, and Mark W. Eshoo-1 1-Ibis Biosciences, 1896 Rutherford Road, Carlsbad, CA 92008 (Mark W. Eshoo e-mail: meshoo@ibisbio.com), 2-University of Medicine and Dentistry of New Jersey, Dept. of Medicine, 185 South Orange Ave., Newark, NJ 07103.]

ABSTRACT: Ticks harbor numerous bacterial, protozoal, and viral pathogens that can cause serious infections in humans and domestic animals. Active surveillance of the tick vector can provide insight into the frequency and distribution of important pathogens in the environment. Nucleic-acid based detection of tick-borne bacterial, protozoan, and viral pathogens requires the extraction of both DNA and RNA (total nucleic acids) from ticks. Traditional methods for nucleic acid extraction are limited to extraction of either DNA or the RNA from a sample. Here we present a simple bead-beating based protocol for extraction of DNA and RNA from a single tick and show detection of Borrelia burgdorferi and Powassan virus from individual, infected Ixodes scapularis ticks. We determined expected yields for total nucleic acids by this protocol for a variety of adult tick species. The method is applicable to a variety of arthropod vectors, including fleas and mosquitoes, and was partially automated on a liquid handling robot.


23. PLoS ONE 5(5) 2010 http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0010650 Genotypic variation and Mixtures of Lyme Borrelia in Ixodes Ticks from North America and Europe
Chris D. Crowder, Heather E. Matthews, Megan A. Rounds, Curtis A. Phillipson, Feng Li, Ranga Sampath, David J. Ecker, Mark W. Eshoo* – Ibis Biosciences, Carlsbad, California, United States of America; Scott R. Campbell – Suffolk County Department of Health Services, Yaphank, New York, United States of America; Benjamin J. Luft – Department of Medicine, State University of New York at Stony Brook, Stony Brook, New York, United States of America; Oliver Nolte – Laboratory of Dr. Brunner, Constance, Germany; Steven Schutzer – Department of Medicine, University of Medicine and Dentistry of New Jersey, Newark, New Jersey, United States of America *Corresponding Author Email: Mark W. Eshoo meshoo@ibisbio.com

ABSTRACT: Lyme disease, caused by various species of Borrelia, is transmitted by Ixodes ticks in North America and Europe. Studies have shown the genotype of Borrelia burgdorferi sensu stricto (s.s.) or the species of B. burgdorferi sensu lato (s.l.) affects the ability of the bacteria to cause local or disseminated infection in humans. Methodology/Principal Findings: We used a multilocus PCR electrospray mass spectrometry assay to determine the species and genotype Borrelia from ticks collected in New York, Connecticut, Indiana, Southern Germany, and California and characterized isolates from parts of the United States and Europe. These analyses identified 53 distinct genotypes of B. burgdorferi sensu stricto with higher resolution than ospC typing. Genotypes of other members of the B. burgdorferi sensu lato complex were also identified and genotyped including B. afzelii, B. garinii, B. lusitaniae, B. spielmanii, and B. valaisiana. While each site in North America had genotypes unique to that location, we found genotypes shared between individual regions and two genotypes found across the United States. Significant B. burgdorferi s.s. genotypic diversity was observed between North America and Europe: only 6.6% of US genotypes (3 of 45) were found in Europe and 27% of the European genotypes (3 of 11) were observed in the US. Interestingly, 39% of adult Ixodes scapularis ticks from North America were infected with more than one genotype of B. burgdorferi s.s. and 22.2% of Ixodes ricinus ticks from Germany were infected with more than one genotype of B. burgdorferi s.l. Conclusions/Significance: The presence of multiple Borrelia genotypes in ticks increases the probability that a person will be infected with more than one genotype of B. burgdorferi, potentially increasing the risks of disseminated Lyme disease. Our study indicates that the genotypic diversity of Borrelia in ticks in both North America and Europe is higher then previously reported and can have potential clinical consequences.


22. Neurobiology of Disease 2010 http://www.sciencedirect.com/science/article/pii/S0969996109003386 Inflammation and central nervous system Lyme disease.
Brian A. Fallon*-a, d, Elizabeth S. Levin-b, Pernilla J. Schweitzer-b, David Hardestya-c a Department of Psychiatry, Columbia University, New York, NY, USA b College of Physicians and Surgeons, Columbia University, New York, NY, USA c Department of Neurology, Columbia University, New York, NY, USA d New York State Psychiatric Institute, New York, NY, USA

*Corresponding Author, Dr. Fallon, Columbia University Medical Center, 1051 Riverside Drive, New York, NY 10032, United States. Fax: +1 212 543 6515. baf1@columbia.edu

ABSTRACT: Lyme disease, caused by the bacterium Borrelia burgdorferi, can cause multi-systemic signs and symptoms, including peripheral and central nervous system disease. This review examines the evidence for and mechanisms of inflammation in neurologic Lyme disease, with a specific focus on the central nervous system, drawing upon human studies and controlled research with experimentally infected rhesus monkeys. Directions for future human research are suggested that may help to clarify the role of inflammation as a mediator of the chronic persistent symptoms experienced by some patients despite antibiotic treatment for neurologic Lyme disease.


21. Arch Gen Psychiatry. 2009;66(5):554-563. 
http://archpsyc.jamanetwork.com/article.aspx?articleid=483068
Regional Cerebral Blood Flow and Metabolic Rate in Persistent Lyme Encephalopathy
Brian A. Fallon, MD*; Richard B. Lipkin, BA; Kathy M. Corbera, MD; Shan Yu, PhD; Mitchell S. Nobler, MD; John G. Keilp, PhD; Eva Petkova, PhD; Sarah H. Lisanby, MD; James R. Moeller, PhD; Iordan Slavov, PhD; Ronald Van Heertum, MD; Brett D. Mensh, MD, PhD; Harold A. Sackeim, PhD Author Affiliations: Departments of Psychiatry (Drs Fallon, Corbera, Yu, Keilp, Petkova, Lisanby, Moeller, Slavov, Mensh, and Sackeim), Radiology (Drs Van Heertum and Sackeim), and Biostatistics (Dr Petkova), College of Physicians and Surgeons of Columbia University, New York, New York; the New York Sta