TY - JOUR TI - Regulation of Drosophila Lifespan by bellwether Promoter Alleles AU - Garcia, Júlia Frankenberg AU - Carbone, Mary Anna AU - Mackay, Trudy F. C. AU - Anholt, Robert R. H. T2 - Scientific Reports AB - Longevity varies among individuals, but how natural genetic variation contributes to variation in lifespan is poorly understood. Drosophila melanogaster presents an advantageous model system to explore the genetic underpinnings of longevity, since its generation time is brief and both the genetic background and rearing environment can be precisely controlled. The bellwether (blw) gene encodes the α subunit of mitochondrial ATP synthase. Since metabolic rate may influence lifespan, we investigated whether alternative haplotypes in the blw promoter affect lifespan when expressed in a co-isogenic background. We amplified 521 bp upstream promoter sequences containing alternative haplotypes and assessed promoter activity both in vitro and in vivo using a luciferase reporter system. The AG haplotype showed significantly greater expression of luciferase than the GT haplotype. We then overexpressed a blw cDNA construct driven by either the AG or GT haplotype promoter in transgenic flies and showed that the AG haplotype also results in greater blw cDNA expression and a significant decrease in lifespan relative to the GT promoter haplotype, in male flies only. Thus, our results show that naturally occurring regulatory variants of blw affect lifespan in a sex-specific manner. DA - 2017/6/23/ PY - 2017/6/23/ DO - 10.1038/S41598-017-04530-X VL - 7 IS - 1 J2 - Sci Rep LA - en OP - SN - 2045-2322 UR - http://dx.doi.org/10.1038/S41598-017-04530-X DB - Crossref ER - TY - JOUR TI - Tailoring Panchromatic Absorption and Excited-State Dynamics of Tetrapyrrole–Chromophore (Bodipy, Rylene) Arrays—Interplay of Orbital Mixing and Configuration Interaction AU - Mandal, Amit Kumar AU - Diers, James R. AU - Niedzwiedzki, Dariusz M. AU - Hu, Gongfang AU - Liu, Rui AU - Alexy, Eric J. AU - Lindsey, Jonathan S. AU - Bocian, David F. AU - Holten, Dewey T2 - Journal of the American Chemical Society AB - Three sets of tetrapyrrole–chromophore arrays have been examined that exhibit panchromatic absorption across large portions of the near-ultraviolet (NUV) to near-infrared (NIR) spectrum along with favorable excited-state properties for use in solar-energy conversion. The arrays vary the tetrapyrrole (porphyrin, chlorin, bacteriochlorin), chromophore (boron-dipyrrin, perylene, terrylene), and attachment sites (meso-position, β-pyrrole position). In all, seven dyads, one triad, and nine benchmarks in toluene and benzonitrile were studied using steady-state and time-resolved absorption and fluorescence spectroscopy. The results were analyzed with the aid of density functional theory (DFT) and time-dependent DFT calculations. Natural transition orbitals (NTOs) were constructed to assess the net change in electron density associated with each NUV–NIR absorption transition. The porphyrin–perylene dyad P-PMI displays the most even spectral coverage from 400 to 700 nm, with an average ε ∼ 43 000 M–1 cm–1. A significant contributor is a chromophore-induced reduction in the configuration interaction involving the four frontier molecular orbitals of benchmark porphyrins and associated constructive/destructive transition-dipole interference that results in intense (ε ∼ 400 000 M–1 cm–1) NUV and weak (<20 000 M–1 cm–1) visible features. P-PMI has an S1 lifetime (τS) of 4.7 ns in toluene and 1.3 ns in benzonitrile. Bacteriochlorin analogue BC-PMI has more extended spectral coverage (350–750 nm) and τS = 2.8 ns in toluene and 30 ps in benzonitrile. Terrylene analogue P-TMI has intermediate optical characteristics with τS = 310 ps in toluene and 150 ps in benzonitrile. The NTOs for most arrays show that S0 → S1 primarily involves the tetrapyrrole, but for P-TMI the NTOs have electron density delocalized over the two units as a result of extensive orbital mixing. Collectively, the insights obtained should aid the design of tetrapyrrole-based architectures for panchromatic light-harvesting systems for solar-energy conversion. DA - 2017/11/21/ PY - 2017/11/21/ DO - 10.1021/JACS.7B09548 VL - 139 IS - 48 SP - 17547-17564 J2 - J. Am. Chem. Soc. LA - en OP - SN - 0002-7863 1520-5126 UR - http://dx.doi.org/10.1021/JACS.7B09548 DB - Crossref ER - TY - JOUR TI - Tracking 19th Century Late Blight from Archival Documents using Text Analytics and Geoparsing AU - Tateosian, L. AU - Guenter, R. AU - Yang, Y.P. AU - Ristaino, J. T2 - Free and Open Source Software for Geospatial (FOSS4G) Conference Proceedings DA - 2017/// PY - 2017/// DO - 10.7275/R5J964K5 VL - 17 IS - 1 SP - 17 UR - http://scholarworks.umass.edu/foss4g/vol17/iss1/17/ ER - TY - JOUR TI - An inclusive Research Education Community (iREC): Impact of the SEA-PHAGES program on research outcomes and student learning AU - Hanauer, David I. AU - Graham, Mark J. AU - Betancur, Laura AU - Bobrownicki, Aiyana AU - Cresawn, Steven G. AU - Garlena, Rebecca A. AU - Jacobs-Sera, Deborah AU - Kaufmann, Nancy AU - Pope, Welkin H. AU - Russell, Daniel A. AU - Jacobs, William R., Jr. AU - Sivanathan, Viknesh AU - Asai, David J. AU - Hatfull, Graham F. T2 - Proceedings of the National Academy of Sciences AB - Engaging undergraduate students in scientific research promises substantial benefits, but it is not accessible to all students and is rarely implemented early in college education, when it will have the greatest impact. An inclusive Research Education Community (iREC) provides a centralized scientific and administrative infrastructure enabling engagement of large numbers of students at different types of institutions. The Science Education Alliance-Phage Hunters Advancing Genomics and Evolutionary Science (SEA-PHAGES) is an iREC that promotes engagement and continued involvement in science among beginning undergraduate students. The SEA-PHAGES students show strong gains correlated with persistence relative to those in traditional laboratory courses regardless of academic, ethnic, gender, and socioeconomic profiles. This persistent involvement in science is reflected in key measures, including project ownership, scientific community values, science identity, and scientific networking. DA - 2017/12/5/ PY - 2017/12/5/ DO - 10.1073/pnas.1718188115 VL - 114 IS - 51 SP - 13531-13536 J2 - Proc Natl Acad Sci USA LA - en OP - SN - 0027-8424 1091-6490 UR - http://dx.doi.org/10.1073/pnas.1718188115 DB - Crossref KW - bacteriophage KW - genomics KW - science education KW - evolution KW - assessment ER - TY - JOUR TI - Foundational and Translational Research Opportunities to Improve Plant Health AU - Michelmore, Richard AU - Coaker, Gitta AU - Bart, Rebecca AU - Beattie, Gwyn AU - Bent, Andrew AU - Bruce, Toby AU - Cameron, Duncan AU - Dangl, Jeffery AU - Dinesh-Kumar, Savithramma AU - Edwards, Rob AU - Eves-van den Akker, Sebastian AU - Gassmann, Walter AU - Greenberg, Jean T. AU - Hanley-Bowdoin, Linda AU - Harrison, Richard J. AU - Harvey, Jagger AU - He, Ping AU - Huffaker, Alisa AU - Hulbert, Scot AU - Innes, Roger AU - Jones, Jonathan D. G. AU - Kaloshian, Isgouhi AU - Kamoun, Sophien AU - Katagiri, Fumiaki AU - Leach, Jan AU - Ma, Wenbo AU - McDowell, John AU - Medford, June AU - Meyers, Blake AU - Nelson, Rebecca AU - Oliver, Richard AU - Qi, Yiping AU - Saunders, Diane AU - Shaw, Michael AU - Smart, Christine AU - Subudhi, Prasanta AU - Torrance, Lesley AU - Tyler, Bret AU - Valent, Barbara AU - Walsh, John T2 - Molecular Plant-Microbe Interactions AB - The white paper reports the deliberations of a workshop focused on biotic challenges to plant health held in Washington, D.C. in September 2016. Ensuring health of food plants is critical to maintaining the quality and productivity of crops and for sustenance of the rapidly growing human population. There is a close linkage between food security and societal stability; however, global food security is threatened by the vulnerability of our agricultural systems to numerous pests, pathogens, weeds, and environmental stresses. These threats are aggravated by climate change, the globalization of agriculture, and an over-reliance on nonsustainable inputs. New analytical and computational technologies are providing unprecedented resolution at a variety of molecular, cellular, organismal, and population scales for crop plants as well as pathogens, pests, beneficial microbes, and weeds. It is now possible to both characterize useful or deleterious variation as well as precisely manipulate it. Data-driven, informed decisions based on knowledge of the variation of biotic challenges and of natural and synthetic variation in crop plants will enable deployment of durable interventions throughout the world. These should be integral, dynamic components of agricultural strategies for sustainable agriculture. DA - 2017/7// PY - 2017/7// DO - 10.1094/mpmi-01-17-0010-cr VL - 30 IS - 7 SP - 515-516 J2 - MPMI LA - en OP - SN - 0894-0282 UR - http://dx.doi.org/10.1094/mpmi-01-17-0010-cr DB - Crossref ER - TY - JOUR TI - Quantitation of Tolyporphins, Diverse Tetrapyrrole Secondary Metabolites with Chlorophyll-Like Absorption, from a Filamentous Cyanobacterium-Microbial Community AU - Zhang, Yunlong AU - Zhang, Ran AU - Hughes, Rebecca-Ayme AU - Dai, Jingqiu AU - Gurr, Joshua R. AU - Williams, Philip G. AU - Miller, Eric S. AU - Lindsey, Jonathan S. T2 - Phytochemical Analysis AB - Abstract Introduction Tolyporphins are unusual tetrapyrrole macrocycles produced by a non‐axenic filamentous cyanobacterium (HT‐58‐2). Tolyporphins A–J, L, and M share a common dioxobacteriochlorin core, differ in peripheral substituents, and exhibit absorption spectra that overlap that of the dominant cyanobacterial pigment, chlorophyll a . Identification and accurate quantitation of the various tolyporphins in these chlorophyll‐rich samples presents challenges. Objective To develop methods for the quantitative determination of tolyporphins produced under various growth conditions relative to that of chlorophyll a . Methodology Chromatographic fractionation of large‐scale (440 L) cultures afforded isolated individual tolyporphins. Lipophilic extraction of small‐scale (25 mL) cultures, HPLC separation with an internal standard, and absorption detection enabled quantitation of tolyporphin A and chlorophyll a , and by inference the amounts of tolyporphins A–M. Absorption spectroscopy with multicomponent analysis of lipophilic extracts (2 mL cultures) afforded the ratio of all tolyporphins to chlorophyll a . The reported absorption spectral data for the various tolyporphins required re‐evaluation for quantitative purposes. Results and Discussion The amount of tolyporphin A after 50 days of illumination ranged from 0.13 nmol/mg dry cells (media containing nitrate) to 1.12 nmol/mg (without nitrate), with maximum 0.23 times that of chlorophyll a . Under soluble‐nitrogen deprivation after 35–50 days, tolyporphin A represents 1/3–1/2 of the total tolyporphins, and the total amount of tolyporphins is up to 1.8‐fold that of chlorophyll a . Conclusions The quantitative methods developed herein should facilitate investigation of the biosynthesis of tolyporphins (and other tetrapyrroles) as well as examination of other strains for production of tolyporphins. Copyright © 2017 John Wiley & Sons, Ltd. DA - 2017/11/6/ PY - 2017/11/6/ DO - 10.1002/pca.2735 VL - 29 IS - 2 SP - 205-216 J2 - Phytochem. Anal. LA - en OP - SN - 0958-0344 UR - http://dx.doi.org/10.1002/pca.2735 DB - Crossref KW - HPLC KW - mass spectrometry KW - absorption spectroscopy with multicomponent analysis KW - nitrogen deprivation KW - growth curve ER - TY - CONF TI - Tracking 19th century late blight from archival documents using text analytics and geoparsing AU - Tateosian, Laura AU - Guenter, Rachael AU - Yang, Yi-Peng AU - Ristaino, Jean C2 - 2017/// C3 - Free and Open Source Software for Geospatial (FOSS4G) Conference Proceedings DA - 2017/// VL - 17 SP - 17 M1 - 1 ER - TY - RPRT TI - A Risk Analysis of precision farming for tomato production AU - Liu, Yangxuan AU - Langemeier, Michael AU - Small, Ian AU - Joseph, Laura AU - Fry, William AU - Ristaino, Jean AU - Saville, Amanda DA - 2017/// PY - 2017/// ER - TY - CONF TI - Evolutionary relatedness and sources of US lineages of Phytophthora infestans (Mont.) de Bary. AU - Saville, Amanda C AU - Ristaino, Jean T2 - APSNET C2 - 2017/// C3 - 2017 APS Annual Meeting DA - 2017/// ER - TY - CONF TI - Charles Darwin and the Irish Potato Famine:“A Painfully Interesting Subject” AU - Ristaino, Jean B AU - Pfister, Donald T2 - APSNET C2 - 2017/// C3 - 2017 APS Annual Meeting DA - 2017/// ER - TY - CONF TI - A species of Pestalotiopsis identified infecting red mangrove in The Bahamas AU - Rossi, Ryann E AU - Layman, Craig A AU - Ristaino, Jean B T2 - APSNET C2 - 2017/// C3 - 2017 APS Annual Meeting DA - 2017/// ER - TY - JOUR TI - Bartonella henselae, Bartonella koehlerae and Rickettsia rickettsii seroconversion and seroreversion in a dog with acute-onset fever, lameness, and lymphadenopathy followed by a protracted disease course AU - Golly, Elizabeth AU - Breitschwerdt, Edward B. AU - Balakrishnan, Nandhakumar AU - Moore, Deedee AU - Bizikova, Petra T2 - Veterinary Parasitology: Regional Studies and Reports AB - Following recent tick exposure in Arkansas, a 2-year-old, female spayed Labradoodle was examined because of a one-week history of lethargy and shifting-leg lameness. The dog was febrile, had prominent lymph nodes, dull mentation, a stiff gait, and left forelimb lameness. Thrombocytopenia was the only initial hematological or biochemical abnormality. Despite treatment with doxycycline for suspected Rocky Mountain spotted fever, the dog continued to have waxing and waning clinical signs including inappetence, fever, shifting-leg lameness, lymphadenopathy, splenomegaly, and weight loss in association with moderate to severe hematological abnormalities, including anemia, thrombocytopenia, neutrophilia, and monocytosis. Sequential serological testing confirmed Bartonella henselae, Bartonella koehlerae and R. rickettsii seroconversion. Doxycycline, enrofloxacin and clarithromycin were administered in sequential combination for treatment of rickettsioses, B. henselae and B. koehlerae. Prednisone, thyroid supplementation and other drugs were administered to elicit symptomatic improvement. Based upon seroreversion, and the eventual resolution of all clinical and hematological abnormalities, therapeutic elimination of all three pathogens was seemingly achieved. Whether cortisol insufficiency due to adrenal exhaustion syndrome or post-infectious immune-mediated sequelae contributed to the symptoms and pathophysiological abnormalities reported in this dog was not determined, but are considerations for future cases. DA - 2017/1// PY - 2017/1// DO - 10.1016/j.vprsr.2016.12.002 VL - 7 SP - 19-24 J2 - Veterinary Parasitology: Regional Studies and Reports LA - en OP - SN - 2405-9390 UR - http://dx.doi.org/10.1016/j.vprsr.2016.12.002 DB - Crossref ER - TY - JOUR TI - Improved molecular detection of Babesia infections in animals using a novel quantitative real-time PCR diagnostic assay targeting mitochondrial DNA AU - Qurollo, Barbara A. AU - Archer, Nikole R. AU - Schreeg, Megan E. AU - Marr, Henry S. AU - Birkenheuer, Adam J. AU - Haney, Kaitlin N. AU - Thomas, Brittany S. AU - Breitschwerdt, Edward B. T2 - Parasites & Vectors AB - Babesiosis is a protozoal, tick transmitted disease found worldwide in humans, wildlife and domesticated animals. Commonly used approaches to diagnose babesiosis include microscopic examination of peripheral blood smears, detection of circulating antibodies and PCR. To screen and differentiate canine Babesia infections many PCR assays amplify the 18S rRNA gene. These sequences contain hypervariable regions flanked by highly conserved regions allowing for amplification of a broad-range of Babesia spp. However, differences in the 18S rRNA gene sequence of distantly related clades can make it difficult to design assays that will amplify all Babesia species while excluding the amplification of other eukaryotes. By targeting Babesia mitochondrial genome (mtDNA), we designed a novel three primer qPCR with greater sensitivity and broader screening capabilities to diagnose and differentiate Babesia spp. Using 13 Babesia mtDNA sequences, a region spanning two large subunit rRNA gene fragments (lsu5-lsu4) was aligned to design three primers for use in a qPCR assay (LSU qPCR) capable of amplifying a wide range of Babesia spp. Plasmid clones were generated and used as standards to determine efficiency, linear dynamic range and analytical sensitivity. Animals naturally infected with vector-borne pathogens were tested retrospectively and prospectively to determine relative clinical sensitivity and specificity by comparing the LSU qPCR to an established 18S rDNA qPCR. The LSU qPCR efficiencies ranged between 92 and 100% with the limit of detection at five copies/reaction. The assay did not amplify mammalian host or other vector-borne pathogen gDNA except Cytauxzoon felis (a feline protozoal pathogen). The LSU qPCR assay amplified 12 different Babesia. sp. and C. felis from 31/31 (100%) archived samples, whereas the 18S qPCR amplified only 26/31 (83.9%). By prospective analysis, 19/394 diagnostic accessions (4.8%) were LSU qPCR positive, compared to 11/394 (2.8%) 18S rDNA qPCR positive. We have developed a more sensitive qPCR assay with a more expansive range of Babesia spp. detection by targeting a highly conserved region of mtDNA, when compared to an established 18S qPCR. DA - 2017/3/7/ PY - 2017/3/7/ DO - 10.1186/s13071-017-2064-1 VL - 10 IS - 1 J2 - Parasites Vectors LA - en OP - SN - 1756-3305 UR - http://dx.doi.org/10.1186/s13071-017-2064-1 DB - Crossref KW - Canine babesiosis KW - Babesia KW - Mitochondrial DNA KW - Quantitative PCR ER - TY - JOUR TI - Sensitivity and specificity levels of two rapid assays for antibodies to Anaplasma spp. in dogs AU - Liu, Jiayou AU - Eberts, Matthew AU - Bewsey, Hannah AU - O’Connor, Thomas P. AU - Chandrashekar, Ramaswamy AU - Breitschwerdt, Edward B. T2 - Journal of Veterinary Diagnostic Investigation AB - Canine anaplasmosis is a tick-borne disease of dogs that results following infection with Anaplasma phagocytophilum or Anaplasma platys. The SNAP 4Dx Plus test (IDEXX Laboratories) and the VetScan Canine Anaplasma Rapid test (Abaxis) are commercial in-house rapid tests for the detection of antibody to these 2 antigenically related Anaplasma species. We evaluated 2 tests using serum and whole blood samples obtained from reference laboratories and veterinary hospitals. Samples were obtained from regions of the country known to be habitats of the primary tick vectors. The A. phagocytophilum sample set comprised 236 dog sera from the northeastern and midwestern United States; the A. platys sample set comprised 179 sera from dogs living in the southwestern United States. An indirect immunofluorescent antibody (IFA) test and an A. platys species-specific ELISA were used as reference assays for the A. phagocytophilum and A. platys samples, respectively. The SNAP test demonstrated significantly higher sensitivity (84.7% for A. phagocytophilum and 83.1% for A. platys), compared to the VetScan test (39.0% for A. phagocytophilum and 57.6% for A. platys). The specificity of the SNAP test (95.8% for A. phagocytophilum and 99.2% for A. platys) was significantly greater than the VetScan test (85.6% for A. phagocytophilum and 82.5% for A. platys). In a separate clinic study, conducted within an A. phagocytophilum–endemic state (Minnesota) using 154 whole blood samples from client-owned dogs, the VetScan test was negative for 22 of 39 SNAP and IFA seropositive samples. DA - 2017/12/4/ PY - 2017/12/4/ DO - 10.1177/1040638717745932 VL - 30 IS - 2 SP - 290-293 J2 - J VET Diagn Invest LA - en OP - SN - 1040-6387 1943-4936 UR - http://dx.doi.org/10.1177/1040638717745932 DB - Crossref KW - Anaplasma phagocytophilum KW - dogs KW - tick-borne diseases ER - TY - JOUR TI - Doxycycline treatment efficacy in dogs with naturally occurring Anaplasma phagocytophilum infection AU - Yancey, C. B. AU - Diniz, P. P. V. P. AU - Breitschwerdt, E. B. AU - Hegarty, B. C. AU - Wiesen, C. AU - Qurollo, B. A. T2 - Journal of Small Animal Practice AB - Objectives To evaluate doxycycline treatment efficacy and post‐treatment pathogen persistence in dogs naturally infected with Anaplasma phagocytophilum in endemic regions of the USA. Materials and Methods Symptomatic dogs in four US states (MN, WI, CT and CA) were evaluated before treatment with doxycycline and approximately 30 and 60 days post‐treatment. Clinicopathological parameters, co‐exposures and A. phagocytophilum DNA in whole blood and lymph node samples were compared between A. phagocytophilum infected and uninfected dogs. Results In total, 42 dogs fulfilled the inclusion criteria, with 16 dogs (38%) blood PCR‐positive and 26 dogs (62%) blood PCR‐negative for A. phagocytophilum . At initial evaluation, the proportion of clinicopathological abnormalities was similar between A. phagocytophilum infected and uninfected dogs, although thrombocytopenia and lymphopenia were statistically more prevalent among A. phagocytophilum infected dogs. Treatment with doxycycline resulted in resolution of all clinical abnormalities in infected dogs; four dogs had persistent haematological abnormalities, including mild leukopenia, eosinopenia and lymphopenia. All 16 infected dogs became blood PCR‐negative approximately 30 and 60 days after treatment onset. Additionally, 13/13 (100%) lymph node specimens tested post‐treatment were PCR‐negative. Select clinicopathological abnormalities persisted in uninfected dogs after treatment. Clinical Significance The results of this study support the efficacy of doxycycline therapy for clinical treatment of dogs naturally infected with A. phagocytophilum in the USA. This study did not find clinical, haematological or microbiological indicators that supported the persistence of A. phagocytophilum infection in naturally infected dogs following treatment with doxycycline for 28 days. DA - 2017/12/27/ PY - 2017/12/27/ DO - 10.1111/jsap.12799 VL - 59 IS - 5 SP - 286-293 J2 - J Small Anim Pract LA - en OP - SN - 0022-4510 UR - http://dx.doi.org/10.1111/jsap.12799 DB - Crossref ER - TY - JOUR TI - Bartonella Seroepidemiology in Dogs from North America, 2008-2014 AU - Lashnits, E. AU - Correa, M. AU - Hegarty, B.C. AU - Birkenheuer, A. AU - Breitschwerdt, E.B. T2 - Journal of Veterinary Internal Medicine AB - Improved understanding of Bartonella species seroepidemiology in dogs may aid clinical decision making and enhance current understanding of naturally occurring arthropod vector transmission of this pathogen.To identify demographic groups in which Bartonella exposure may be more likely, describe spatiotemporal variations in Bartonella seroreactivity, and examine co-exposures to other canine vector-borne diseases (CVBD).A total of 15,451 serology specimens from dogs in North America were submitted to the North Carolina State University, College of Veterinary Medicine Vector Borne Disease Diagnostic Laboratory between January 1, 2008, and December 31, 2014.Bartonella henselae, Bartonella koehlerae, and Bartonella vinsonii subspecies berkhoffii indirect fluorescent antibody (IFA) serology results, as well as results from a commercial assay kit screening for Dirofilaria immitis antigen and Ehrlichia species, Anaplasma phagocytophilum, and Borrelia burgdorferi antibodies, and Ehrlichia canis, Babesia canis, Babesia gibsoni, and Rickettsia species IFA results were reviewed retrospectively.Overall, 3.26% of dogs were Bartonella spp. seroreactive; B. henselae (2.13%) and B. koehlerae (2.39%) were detected more frequently than B. vinsonii subsp. berkhoffii (1.42%, P < 0.0001). Intact males had higher seroreactivity (5.04%) than neutered males (2.87%, P < 0.0001) or intact or spayed females (3.22%, P = 0.0003). Mixed breed dogs had higher seroreactivity (4.45%) than purebred dogs (3.02%, P = 0.0002). There was no trend in seasonal seroreactivity; geographic patterns supported broad distribution of exposure, and co-exposure with other CVBD was common.Bartonella spp. exposure was documented throughout North America and at any time of year. Male intact dogs, mixed breed dogs, and dogs exposed to other CVBD have higher seroreactivity to multiple Bartonella species. DA - 2017/12/2/ PY - 2017/12/2/ DO - 10.1111/jvim.14890 VL - 32 IS - 1 SP - 222-231 J2 - J Vet Intern Med LA - en OP - SN - 0891-6640 UR - http://dx.doi.org/10.1111/jvim.14890 DB - Crossref KW - Canine KW - Seroreactivity KW - Zoonoses ER - TY - CONF TI - Polynomial Time Interactive Proofs for Linear Algebra with Exponential Matrix Dimensions and Scalars Given by Polynomial Time Circuits AU - Dumas, Jean-Guillaume AU - Kaltofen, Erich L. AU - Villard, Gilles AU - Zhi, Lihong T2 - the 2017 ACM AB - We present an interactive probabilistic proof protocol that certifies in (log N)O(1) arithmetic and Boolean operations for the verifier the determinant, for example, of an N x N matrix over a field whose entries a(i,j) are given by a single (log NO(1)-depth arithmetic circuit, which contains (log NO(1) field constants and which is polynomial time uniform, for example, which has size (log NO(1). The prover can produce the interactive certificate within a (log NO(1) factor of the cost of computing the determinant. Our protocol is a version of the proofs for muggles protocol by Goldwasser, Kalai and Rothblum [STOC 2008, J. ACM 2015]. An application is the following: suppose in a system of k homogeneous polynomials of total degree ≤ d in the k variables y1,...,yk the coefficient of the term y1e1 ... ykek in the i-th polynomial is the (hypergeometric) value ((i+e1 + ... + ek)!)/((i!)(e1!)...(ek!)), where e! is the factorial of e. Then we have a probabilistic protocol that certifies (projective) solvability or inconsistency of such a system in (k log(d))O(1) bit complexity for the verifier, that is, in polynomial time in the number of variables k and the logarithm of the total degree, log(d). C2 - 2017/// C3 - Proceedings of the 2017 ACM on International Symposium on Symbolic and Algebraic Computation - ISSAC '17 DA - 2017/// DO - 10.1145/3087604.3087640 PB - ACM Press SN - 9781450350648 UR - http://dx.doi.org/10.1145/3087604.3087640 DB - Crossref ER - TY - CONF TI - Early Termination in Parametric Linear System Solving and Rational Function Vector Recovery with Error Correction AU - Kaltofen, Erich L. AU - Pernet, Clément AU - Storjohann, Arne AU - Waddell, Cleveland T2 - the 2017 ACM AB - Consider solving a black box linear system, A(u) x = b(u), where the entries are polynomials in u over a field K, and A(u) is full rank. The solution, x = 1/g(u) f(u), where g is always the least common monic denominator, can be found by evaluating the system at distinct points ξl in K. The solution can be recovered even if some evaluations are erroneous. In [Boyer and Kaltofen, Proc. SNC 2014] the problem is solved with an algorithm that generalizes Welch/Berlekamp decoding of an algebraic Reed-Solomon code. Their algorithm requires the sum of a degree bound for the numerators plus a degree bound for the denominator of the solution. It is possible that the degree bounds input to their algorithm grossly overestimate the actual degrees. We describe an algorithm that given the same inputs uses possibly fewer evaluations to compute the solution. We introduce a second count for the number of evaluations required to recover the solution based on work by Stanley Cabay. The Cabay count includes bounds for the highest degree polynomial in the coefficient matrix and right side vector, but does not require solution degree bounds. Instead our algorithm iterates until the Cabay termination criterion is reached. At this point our algorithm returns the solution. Assuming we have the actual degrees for all necessary input parameters, we give the criterion that determines when the Cabay count is fewer than the generalized Welch/Berlekamp count. C2 - 2017/// C3 - Proceedings of the 2017 ACM on International Symposium on Symbolic and Algebraic Computation - ISSAC '17 DA - 2017/// DO - 10.1145/3087604.3087645 PB - ACM Press SN - 9781450350648 UR - http://dx.doi.org/10.1145/3087604.3087645 DB - Crossref ER - TY - JOUR TI - Spontaneous activation of a MAVS-dependent antiviral signaling pathway determines high basal interferon-beta expression in cardiac myocytes AU - Rivera-Serrano, E. E. AU - DeAngelis, N. AU - Sherry, B. T2 - Journal of Molecular and Cellular Cardiology AB - Viral myocarditis is a leading cause of sudden death in young adults as the limited turnover of cardiac myocytes renders the heart particularly vulnerable to viral damage. Viruses induce an antiviral type I interferon (IFN-α/β) response in essentially all cell types, providing an immediate innate protection. Cardiac myocytes express high basal levels of IFN-β to help pre-arm them against viral infections, however the mechanism underlying this expression remains unclear. Using primary cultures of murine cardiac and skeletal muscle cells, we demonstrate here that the mitochondrial antiviral signaling (MAVS) pathway is spontaneously activated in unstimulated cardiac myocytes but not cardiac fibroblasts or skeletal muscle cells. Results suggest that MAVS association with the mitochondrial-associated ER membranes (MAM) is a determinant of high basal IFN-β expression, and demonstrate that MAVS is essential for spontaneous high basal expression of IFN-β in cardiac myocytes and the heart. Together, results provide the first mechanism for spontaneous high expression of the antiviral cytokine IFN-β in a poorly replenished and essential cell type. DA - 2017/// PY - 2017/// DO - 10.1016/j.yjmcc.2017.08.008 VL - 111 SP - 102-113 ER - TY - JOUR TI - Optimal seed deployment under climate change using spatial models: Application to loblolly pine in the Southeastern US AU - Farjat, A. AU - Reich, B. J. AU - Guinness, J. AU - Whetten, Ross AU - McKeand, Steven AU - Isik, Fikret T2 - Journal of the American Statistical Association AB - Provenance tests are a common tool in forestry designed to identify superior genotypes for planting at specific locations. The trials are replicated experiments established with seed from parent trees collected from different regions and grown at several locations. In this work, a Bayesian spatial approach is developed for modeling the expected relative performance of seed sources using climate variables as predictors associated with the origin of seed source and the planting site. The proposed modeling technique accounts for the spatial dependence in the data and introduces a separable Matérn covariance structure that provides a flexible means to estimate effects associated with the origin and planting site locations. The statistical model was used to develop a quantitative tool for seed deployment aimed to identify the location of superior performing seed sources that could be suitable for a specific planting site under a given climate scenario. Cross-validation results indicate that the proposed spatial models provide superior predictive ability compared to multiple linear regression methods in unobserved locations. The general trend of performance predictions based on future climate scenarios suggests an optimal assisted migration of loblolly pine seed sources from southern and warmer regions to northern and colder areas in the southern USA. Supplementary materials for this article are available online. DA - 2017/// PY - 2017/// DO - 10.1080/01621459.2017.1292179 VL - 112 IS - 519 SP - 909–920 ER - TY - JOUR TI - Mass spectrometric detection of chlorophyll a and the tetrapyrrole secondary metabolite tolyporphin A in the filamentous cyanobacterium HT-58-2. Approaches to high-throughput screening of intact cyanobacteria AU - Zhang, Yunlong AU - Zhang, Ran AU - Nazari, Milad AU - Bagley, Michael C. AU - Miller, Eric S. AU - Williams, Philip G. AU - Muddiman, David C. AU - Lindsey, Jonathan S. T2 - JOURNAL OF PORPHYRINS AND PHTHALOCYANINES AB - Tolyporphins are unusual tetrapyrrole macrocycles produced by the filamentous cyanobacterium–microbial community HT-58-2, the only known source to date. Numerous cyanobacterial samples have been collected worldwide but most have not been screened for secondary metabolites. Identification of tolyporphins typically has entailed lipophilic extraction followed by chromatographic fractionation and spectroscopic and/or mass spectrometric analysis. For quantitation, lengthy lipophilic extraction, sample processing and HPLC separation are needed. Examination by MALDI-TOF-MS (with the matrix 1,5-diaminonaphthalene) of lipophilic crude extracts of small-scale HT-58-2 samples (2 mL) without chromatographic fractionation enabled semi-quantitation of tolyporphin A over a 41-day growth period. Screening for tolyporphin A in intact or slightly sheared and vortexed HT-58-2 samples (no lipophilic extraction), and confirmation of identity by tandem MS, were carried out by IR-MALDESI-FTMS. Tolyporphin A was identified by the molecular ion and four characteristic fragments. The molecular ion of chlorophyll [Formula: see text] also was observed. The sheared and vortexed sample contained substantial numbers of intact cells as demonstrated by regrowth of the filamentous cyanobacterium–microbial culture. The semi-quantitative and rapid qualitative methods developed herein should facilitate examination of other tolyporphin-producing organisms among the vast worldwide strains of cyanobacteria as well as investigation of the biosynthesis of tolyporphins. DA - 2017/11// PY - 2017/11// DO - 10.1142/s108842461750078x VL - 21 IS - 11 SP - 759-768 SN - 1099-1409 KW - IR-MALDESI-FTMS KW - MS/MS KW - MALDI-TOF-MS KW - chlorophyll KW - bacteriochlorin KW - high-throughput screening ER - TY - JOUR TI - Correspondence of Loblolly Pine Response for Fusiform Rust Disease from Local and Wide-Ranging Tests in the Southern United States AU - Spitzer, Jesse AU - Isik, Fikret AU - Whetten, Ross W. AU - Farjat, Alfredo E. AU - McKeand, Steven E. T2 - FOREST SCIENCE AB - Fusiform rust is the most economically important disease of loblolly pine (Pinus taeda L.) in the southern United States. Estimates of family resistance to rust are critical for deployment decisions because 95% of loblolly pine plantations are established with individual families. If families show significant interactions with different pathogen inocula, then the performance of some families in different regions may not be predictable. This study compared rust breeding values of 56 loblolly families estimated from two independent sets of trials. We regressed the rust incidence breeding values of the families estimated from broadly based field tests on breeding values of the same families estimated from narrowly based tests. The model F test was highly significant (P < 0.0001), and breeding values based on local testing explained 75% of the variation in breeding values based on wide-range geographic testing, indicating that local rust breeding values are relatively reliable predictors of families' performance across a broad range of sites. Family rankings were highly consistent across test sites within broadly and narrowly based testing schemes as shown by type B genetic correlations (0.90 and 0.91). We conclude that field testing provides a reliable prediction of the operational value of loblolly families for deployment in regions with a high hazard for fusiform rust. Management and Policy Implications When choosing loblolly pine families to be planted on sites where resistance to fusiform rust is necessary, foresters and landowners can have confidence in the performance data coming from a range of field trials. Results show that local testing for rust resistance in relatively narrow geographic regions provides reasonably reliable rust disease resistance/susceptibility predictions and adequate predictive power for deployment decisions across a broad range of planting sites in the Gulf and Atlantic Coastal Plain regions of the southeastern United States. DA - 2017/10/11/ PY - 2017/10/11/ DO - 10.5849/fs-2016-093r1 VL - 63 IS - 5 SP - 496-503 SN - 1938-3738 KW - tree improvement KW - loblolly KW - pine KW - fusiform KW - rust ER - TY - JOUR TI - The Arabidopsis Leucine-Rich Repeat Receptor Kinase BIR3 Negatively Regulates BAK1 Receptor Complex Formation and Stabilizes BAK1 AU - Imkampe, Julia AU - Halter, Thierry AU - Huang, Shuhua AU - Schulze, Sarina AU - Mazzotta, Sara AU - Schmidt, Nikola AU - Manstretta, Raffaele AU - Postel, Sandra AU - Wierzba, Micheal AU - Yang, Yong AU - Dongen, Walter M. A. M. AU - Stahl, Mark AU - Zipfel, Cyril AU - Goshe, Michael B. AU - Clouse, Steven AU - Vries, Sacco C. AU - Tax, Frans AU - Wang, Xiaofeng AU - Kemmerling, Birgit T2 - PLANT CELL AB - BAK1 is a coreceptor and positive regulator of multiple ligand binding leucine-rich repeat receptor kinases (LRR-RKs) and is involved in brassinosteroid (BR)-dependent growth and development, innate immunity, and cell death control. The BAK1-interacting LRR-RKs BIR2 and BIR3 were previously identified by proteomics analyses of in vivo BAK1 complexes. Here, we show that BAK1-related pathways such as innate immunity and cell death control are affected by BIR3 in Arabidopsis thaliana BIR3 also has a strong negative impact on BR signaling. BIR3 directly interacts with the BR receptor BRI1 and other ligand binding receptors and negatively regulates BR signaling by competitive inhibition of BRI1. BIR3 is released from BAK1 and BRI1 after ligand exposure and directly affects the formation of BAK1 complexes with BRI1 or FLAGELLIN SENSING2. Double mutants of bak1 and bir3 show spontaneous cell death and constitutive activation of defense responses. BAK1 and its closest homolog BKK1 interact with and are stabilized by BIR3, suggesting that bak1 bir3 double mutants mimic the spontaneous cell death phenotype observed in bak1 bkk1 mutants via destabilization of BIR3 target proteins. Our results provide evidence for a negative regulatory mechanism for BAK1 receptor complexes in which BIR3 interacts with BAK1 and inhibits ligand binding receptors to prevent BAK1 receptor complex formation. DA - 2017/9// PY - 2017/9// DO - 10.1105/tpc.17.00376 VL - 29 IS - 9 SP - 2285-2303 SN - 1532-298X ER - TY - JOUR TI - Genomic Analysis of the DNA Replication Timing Program during Mitotic S Phase in Maize (Zea mays) Root Tips AU - Wear, Emily E. AU - Song, Jawon AU - Zynda, Gregory J. AU - LeBlanc, Chantal AU - Lee, Tae-Jin AU - Mickelson-Young, Leigh AU - Concia, Lorenzo AU - Mulvaney, Patrick AU - Szymanski, Eric S. AU - Allen, George C. AU - Martienssen, Robert A. AU - Vaughn, Matthew W. AU - Hanley-Bowdoin, Linda AU - Thompson, William F. T2 - PLANT CELL AB - All plants and animals must replicate their DNA, using a regulated process to ensure that their genomes are completely and accurately replicated. DNA replication timing programs have been extensively studied in yeast and animal systems, but much less is known about the replication programs of plants. We report a novel adaptation of the “Repli-seq” assay for use in intact root tips of maize (Zea mays) that includes several different cell lineages and present whole-genome replication timing profiles from cells in early, mid, and late S phase of the mitotic cell cycle. Maize root tips have a complex replication timing program, including regions of distinct early, mid, and late S replication that each constitute between 20 and 24% of the genome, as well as other loci corresponding to ∼32% of the genome that exhibit replication activity in two different time windows. Analyses of genomic, transcriptional, and chromatin features of the euchromatic portion of the maize genome provide evidence for a gradient of early replicating, open chromatin that transitions gradually to less open and less transcriptionally active chromatin replicating in mid S phase. Our genomic level analysis also demonstrated that the centromere core replicates in mid S, before heavily compacted classical heterochromatin, including pericentromeres and knobs, which replicate during late S phase. DA - 2017/9// PY - 2017/9// DO - 10.1105/tpc.17.00037 VL - 29 IS - 9 SP - 2126-2149 SN - 1532-298X UR - http://europepmc.org/abstract/med/28842533 ER - TY - JOUR TI - Synthesis of tailored hydrodipyrrins and their examination in directed routes to bacteriochlorins and tetradehydrocorrins AU - Zhang, S. F. AU - Reddy, M. N. AU - Mass, O. AU - Kim, H. J. AU - Hu, G. F. AU - Lindsey, Jonathan T2 - New Journal of Chemistry AB - 18 gem-dimethyl stabilized hydrodipyrrins with diverse α-substituents have been prepared and examined in directed syntheses of unsymmetrically substituted hydroporphyrins. DA - 2017/// PY - 2017/// DO - 10.1039/c7nj01892d VL - 41 IS - 19 SP - 11170–11189 ER - TY - JOUR TI - Seroprevalence and risk factors associated with Ehrlichia canis, Anaplasma spp., Borrelia burgdorferi sensu lato, and D. immitis in hunting dogs from southern Italy AU - Piantedosi, Diego AU - Neola, Benedetto AU - D’Alessio, Nicola AU - Di Prisco, Francesca AU - Santoro, Mario AU - Pacifico, Laura AU - Sgroi, Giovanni AU - Auletta, Luigi AU - Buch, Jesse AU - Chandrashekar, Ramaswamy AU - Breitschwerdt, Edward B. AU - Veneziano, Vincenzo T2 - Parasitology Research DA - 2017/8/4/ PY - 2017/8/4/ DO - 10.1007/s00436-017-5574-z VL - 116 IS - 10 SP - 2651-2660 J2 - Parasitol Res LA - en OP - SN - 0932-0113 1432-1955 UR - http://dx.doi.org/10.1007/s00436-017-5574-z DB - Crossref KW - Ehrlichia canis KW - Anaplasma spp KW - Borrelia burgdorferi KW - Dirofilaria immitis KW - Hunting dogs KW - Italy ER - TY - JOUR TI - Photophysical Characterization of the Naturally Occurring Dioxobacteriochlorin Tolyporphin A and Synthetic Oxobacteriochlorin Analogues AU - Hood, Don AU - Niedzwiedzki, Dariusz M. AU - Zhang, Ran AU - Zhang, Yunlong AU - Dai, Jingqiu AU - Miller, Eric S. AU - Bocian, David F. AU - Williams, Philip G. AU - Lindsey, Jonathan S. AU - Holten, Dewey T2 - PHOTOCHEMISTRY AND PHOTOBIOLOGY AB - Abstract Tolyporphins are tetrapyrrole macrocycles produced by a cyanobacterium‐containing culture known as HT ‐58‐2. Tolyporphins A–J are free base dioxobacteriochlorins, whereas tolyporphin K is an oxochlorin. Here, the photophysical characterization is reported of tolyporphin A and two synthetic analogues, an oxobacteriochlorin and a dioxobacteriochlorin. The characterization (in toluene, diethyl ether, ethyl acetate, dichloromethane, 1‐pentanol, 2‐butanone, ethanol, methanol, N,N ‐dimethylformamide and dimethylsulfoxide) includes static absorption and fluorescence spectra, fluorescence quantum yields and time‐resolved data. The data afford the lifetime of the lowest singlet excited state and the yields of the nonradiative decay pathways (intersystem crossing and internal conversion). The three macrocycles exhibit only modest variation in spectroscopic and excited‐state photophysical parameters across the solvents. The long‐wavelength (Q y ) absorption band of tolyporphin A appears at ~680 nm and is remarkably narrow (full‐width‐at‐half‐maximum ~7 nm). The position of the long‐wavelength (Q y ) absorption band of tolyporphin A (~680 nm) more closely resembles that of chlorophyll a (662 nm) than bacteriochlorophyll a (772 nm). The absorption spectra of tolyporphins B–I, K (which were available in minute quantities) are also reported in methanol; the spectra of B–I closely resemble that of tolyporphin A. Taken together, tolyporphin A generally exhibits spectral and photophysical features resembling those of chlorophyll a . DA - 2017/10// PY - 2017/10// DO - 10.1111/php.12781 VL - 93 IS - 5 SP - 1204-1215 SN - 1751-1097 ER - TY - JOUR TI - Pharmacokinetics and efficacy of doxorubicin-loaded plant virus nanoparticles in preclinical models of cancer AU - Madden, Andrew J. AU - Oberhardt, Bruce AU - Lockney, Dustin AU - Santos, Charlene AU - Vennam, Preethi AU - Arney, David AU - Franzen, Stefan AU - Lommel, Steven A. AU - Miller, C. Ryan AU - Gehrig, Paola AU - Zamboni, William C. T2 - NANOMEDICINE AB - To compare the pharmacokinetics and efficacy of doxorubicin containing plant virus nanoparticles (PVNs) with PEGylated liposomal doxorubicin (PLD) and small molecule doxorubicin in two mouse models of cancer.Studies were performed in A375 melanoma and intraperitoneal SKOV3ip1 ovarian cancer xenografts. The PVNs were administered in lower and more frequent doses in the ovarian model.The PVNs were more efficacious than PLD and small molecule doxorubicin in the ovarian cancer model, but not in the melanoma cancer model. The pharmacokinetics profiles of the PVNs showed fast plasma clearance, but more efficient tumor delivery as compared with other carrier-mediated agents.PVNs administered at lower repeated doses provide both pharmacologic and efficacy advantages compared with PLD. DA - 2017/10// PY - 2017/10// DO - 10.2217/nnm-2016-0421 VL - 12 IS - 20 SP - 2519-2532 SN - 1748-6963 KW - efficacy KW - pharmacokinetics KW - plant virus nanoparticles ER - TY - JOUR TI - Linkage map construction and QTL analysis for internal heat necrosis in autotetraploid potato AU - Schumann, Mitchell J. AU - Zeng, Zhao-Bang AU - Clough, Mark E. AU - Yencho, G. Craig T2 - THEORETICAL AND APPLIED GENETICS DA - 2017/10// PY - 2017/10// DO - 10.1007/s00122-017-2941-1 VL - 130 IS - 10 SP - 2045-2056 SN - 1432-2242 ER - TY - JOUR TI - Genome Sequence and Composition of a Tolyporphin-Producing Cyanobacterium-Microbial Community AU - Hughes, Rebecca-Ayme AU - Zhang, Yunlong AU - Zhang, Ran AU - Williams, Philip G. AU - Lindsey, Jonathan S. AU - Miller, Eric S. T2 - APPLIED AND ENVIRONMENTAL MICROBIOLOGY AB - ABSTRACT The cyanobacterial culture HT-58-2 was originally described as a strain of Tolypothrix nodosa with the ability to produce tolyporphins, which comprise a family of distinct tetrapyrrole macrocycles with reported efflux pump inhibition properties. Upon reviving the culture from what was thought to be a nonextant collection, studies of culture conditions, strain characterization, phylogeny, and genomics have been undertaken. Here, HT-58-2 was shown by 16S rRNA analysis to closely align with Brasilonema strains and not with Tolypothrix isolates. Light, fluorescence, and scanning electron microscopy revealed cyanobacterium filaments that are decorated with attached bacteria and associated with free bacteria. Metagenomic surveys of HT-58-2 cultures revealed a diversity of bacteria dominated by Erythrobacteraceae , 97% of which are Porphyrobacter species. A dimethyl sulfoxide washing procedure was found to yield enriched cyanobacterial DNA (presumably by removing community bacteria) and sequence data sufficient for genome assembly. The finished, closed HT-58-2Cyano genome consists of 7.85 Mbp (42.6% G+C) and contains 6,581 genes. All genes for biosynthesis of tetrapyrroles (e.g., heme, chlorophyll a , and phycocyanobilin) and almost all for cobalamin were identified dispersed throughout the chromosome. Among the 6,177 protein-encoding genes, coding sequences (CDSs) for all but two of the eight enzymes for conversion of glutamic acid to protoporphyrinogen IX also were found within one major gene cluster. The cluster also includes 10 putative genes (and one hypothetical gene) encoding proteins with domains for a glycosyltransferase, two cytochrome P450 enzymes, and a flavin adenine dinucleotide (FAD)-binding protein. The composition of the gene cluster suggests a possible role in tolyporphin biosynthesis. IMPORTANCE A worldwide search more than 25 years ago for cyanobacterial natural products with anticancer activity identified a culture (HT-58-2) from Micronesia that produces tolyporphins. Tolyporphins are tetrapyrroles, like chlorophylls, but have several profound structural differences that reside outside the bounds of known biosynthetic pathways. To begin probing the biosynthetic origin and biological function of tolyporphins, our research has focused on studying the cyanobacterial strain, about which almost nothing has been previously reported. We find that the HT-58-2 culture is composed of the cyanobacterium and a community of associated bacteria, complicating the question of which organisms make tolyporphins. Elucidation of the cyanobacterial genome revealed an intriguing gene cluster that contains tetrapyrrole biosynthesis genes and a collection of unknown genes, suggesting that the cluster may be responsible for tolyporphin production. Knowledge of the genome and the gene cluster sharply focuses research to identify related cyanobacterial producers of tolyporphins and delineate the tolyporphin biosynthetic pathway. DA - 2017/10// PY - 2017/10// DO - 10.1128/aem.01068-17 VL - 83 IS - 19 SP - SN - 1098-5336 KW - cyanobacteria KW - genome analysis KW - phylogenetic analysis KW - tetrapyrroles KW - tolyporphin ER - TY - JOUR TI - Disparate gain and loss of parasitic abilities among nematode lineages AU - Holterman, Martijn AU - Karegar, Akbar AU - Mooijman, Paul AU - Megen, Hanny AU - Elsen, Sven AU - Vervoort, Mariette T. W. AU - Quist, Casper W. AU - Karssen, Gerrit AU - Decraemer, Wilfrida AU - Opperman, Charles H. AU - Bird, David M. AU - Kammenga, Jan AU - Goverse, Aska AU - Smant, Geert AU - Helder, Johannes T2 - PLOS ONE AB - Plant parasitism has arisen time and again in multiple phyla, including bacteria, fungi, insects and nematodes. In most of these organismal groups, the overwhelming diversity hampers a robust reconstruction of the origins and diversification patterns of this trophic lifestyle. Being a moderately diversified phylum with ≈ 4,100 plant parasites (15% of total biodiversity) subdivided over four independent lineages, nematodes constitute a major organismal group for which the genesis of plant parasitism could be mapped. Since substantial crop losses worldwide have been attributed to less than 1% of these plant parasites, research efforts are severely biased towards this minority. With the first molecular characterisation of numerous basal and supposedly harmless plant parasites as well as their non-parasitic relatives, we were able to generate a comprehensive molecular framework that allows for the reconstruction of trophic diversification for a complete phylum. In each lineage plant parasites reside in a single taxonomic grouping (family or order), and by taking the coverage of the next lower taxonomic level as a measure for representation, 50, 67, 100 and 85% of the known diversity was included. We revealed distinct gain and loss patterns with regard to plant parasitism per se as well as host exploitation strategies between these lineages. Our map of parasitic nematode biodiversity also revealed an unanticipated time reversal in which the two most ancient lineages showed the lowest level of ecological diversification and vice versa. DA - 2017/9/21/ PY - 2017/9/21/ DO - 10.1371/journal.pone.0185445 VL - 12 IS - 9 SP - SN - 1932-6203 UR - http://www.scopus.com/inward/record.url?eid=2-s2.0-85029856456&partnerID=MN8TOARS ER - TY - JOUR TI - Spotted Gar and the Evolution of Innate Immune Receptors AU - Wcisel, Dustin J. AU - Ota, Tatsuya AU - Litman, Gary W. AU - Yoder, Jeffrey A. T2 - JOURNAL OF EXPERIMENTAL ZOOLOGY PART B-MOLECULAR AND DEVELOPMENTAL EVOLUTION AB - The resolution of the gar genome affords an opportunity to examine the diversification and functional specialization of immune effector molecules at a distant and potentially informative point in phylogenetic development. Although innate immunity is effected by a particularly large number of different families of molecules, the focus here is to provide detailed characterization of several families of innate receptors that are encoded in large multigene families, for which orthologous forms can be identified in other species of bony fish but not in other vertebrate groups as well as those for which orthologs are present in other vertebrate species. The results indicate that although teleost fish and the gar, as a holostean reference species, share gene families thought previously to be restricted to the teleost fish, the manner in which the members of the multigene families of innate immune receptors have undergone diversification is different in these two major phylogenetic radiations. It appears that both the total genome duplication and different patterns of genetic selection have influenced the derivation and stabilization of innate immune genes in a substantial manner during the course of vertebrate evolution. DA - 2017/11// PY - 2017/11// DO - 10.1002/jez.b.22738 VL - 328 IS - 7 SP - 666-684 SN - 1552-5015 ER - TY - JOUR TI - Sequence analysis of RAS and RAF mutation hot spots in canine carcinoma AU - Mochizuki, H. AU - Breen, M. T2 - VETERINARY AND COMPARATIVE ONCOLOGY AB - Recent discovery of the BRAF V595E mutation in a variety of canine cancers indicates that mutant BRAF may represent a novel therapeutic target. Presence of RAS mutations is associated with poor tumour response to BRAF inhibition but has not been investigated in BRAF-mutated canine cancers. The aim of this study was to evaluate the mutational status of three RAS genes (HRAS, KRAS and NRAS) in four types of canine carcinoma with and without the BRAF V595E mutation. Novel HRAS mutations were identified in 18% (3/17) of oral squamous cell carcinoma, whereas 17% (3/18) of pulmonary carcinoma carried KRAS or NRAS mutations. These RAS mutations and BRAF V595E were mutually exclusive, indicating similar functional consequence of these mutations (e.g. MAPK pathway activation). In contrast, RAS mutations were absent in 39 urothelial carcinoma and 19 prostatic carcinoma, adding another rational for BRAF-targeted therapy for these canine cancers. DA - 2017/12// PY - 2017/12// DO - 10.1111/vco.12275 VL - 15 IS - 4 SP - 1598-1605 SN - 1476-5829 KW - bladder KW - cancer KW - dog KW - prostate KW - squamous cell carcinoma KW - transitional cell carcinoma ER - TY - JOUR TI - Reciprocal cross-regulation of VND and SND multigene TF families for wood formation in Populus trichocarpa AU - Lin, Ying-Chung Jimmy AU - Chen, Hao AU - Li, Quanzi AU - Li, Wei AU - Wang, Jack P. AU - Shi, Rui AU - Tunlaya-Anukit, Sermsawat AU - Shuai, Peng AU - Wang, Zhifeng AU - Ma, Hongyan AU - Li, Huiyu AU - Sun, Ying-Hsuan AU - Sederoff, Ronald R. AU - Chiang, Vincent L. T2 - PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA AB - Secondary cell wall (SCW) biosynthesis is the biological process that generates wood, an important renewable feedstock for materials and energy. NAC domain transcription factors, particularly Vascular-Related NAC-Domain (VND) and Secondary Wall-Associated NAC Domain (SND) proteins, are known to regulate SCW differentiation. The regulation of VND and SND is important to maintain homeostasis for plants to avoid abnormal growth and development. We previously identified a splice variant, PtrSND1-A2IR , derived from PtrSND1-A2 as a dominant-negative regulator, which suppresses the transactivation of all PtrSND1 family members. PtrSND1-A2IR also suppresses the self-activation of the PtrSND1 family members except for its cognate transcription factor, PtrSND1-A2, suggesting the existence of an unknown factor needed to regulate PtrSND1-A2 Here, a splice variant, PtrVND6-C1IR , derived from PtrVND6-C1 was discovered that suppresses the protein functions of all PtrVND6 family members. PtrVND6-C1IR also suppresses the expression of all PtrSND1 members, including PtrSND1-A2, demonstrating that PtrVND6-C1IR is the previously unidentified regulator of PtrSND1-A2 We also found that PtrVND6-C1IR cannot suppress the expression of its cognate transcription factor, PtrVND6-C1PtrVND6-C1 is suppressed by PtrSND1-A2IR Both PtrVND6-C1IR and PtrSND1-A2IR cannot suppress their cognate transcription factors but can suppress all members of the other family. The results indicate that the splice variants from the PtrVND6 and PtrSND1 family may exert reciprocal cross-regulation for complete transcriptional regulation of these two families in wood formation. This reciprocal cross-regulation between families suggests a general mechanism among NAC domain proteins and likely other transcription factors, where intron-retained splice variants provide an additional level of regulation. DA - 2017/11/7/ PY - 2017/11/7/ DO - 10.1073/pnas.1714422114 VL - 114 IS - 45 SP - E9722-E9729 SN - 0027-8424 KW - reciprocal cross-regulation KW - NAC transcription factors KW - alternative splicing KW - wood formation KW - Populus trichocarpa ER - TY - JOUR TI - Prevalence of Bartonella spp. by culture, PCR and serology, in veterinary personnel from Spain AU - Oteo, José A. AU - Maggi, Ricardo AU - Portillo, Aránzazu AU - Bradley, Julie AU - García-Álvarez, Lara AU - San-Martín, Montserrat AU - Roura, Xavier AU - Breitschwerdt, Edward T2 - Parasites & Vectors AB - The genus Bartonella includes fastidious, facultative intracellular bacteria mainly transmitted by arthropods and distributed among mammalian reservoirs. Bartonella spp. implicated as etiological agents of zoonoses are increasing. Apart from the classical Bartonella henselae, B. bacilliformis or B. quintana, other species (B. elizabethae, B. rochalimae, B. vinsonii arupensis and B. v. berkhoffii, B. tamiae or B. koehlerae, among others) have also been associated with human and/or animal diseases. Laboratory techniques for diagnosis (culture, PCR assays and serology) usually show lack of sensitivity. Since 2005, a method based on a liquid enrichment Bartonella alphaproteobacteria growth medium (BAPGM) followed by PCRs for the amplification of Bartonella spp. has been developed. We aimed to assess culture, molecular and serological prevalence of Bartonella infections in companion animal veterinary personnel from Spain. Each of 89 participants completed a questionnaire. Immunofluorescence assays (IFA) using B. vinsonii berkhoffii (genotypes I, II and III), B. henselae, B. quintana and B. koehlerae as antigens were performed. A cut-off of 1:64 was selected as a seroreactivity titer. Blood samples were inoculated into BAPGM and subcultured onto blood agar plates. Bartonella spp. was detected using conventional and quantitative real-time PCR assays and DNA sequencing. Among antigens corresponding to six Bartonella spp. or genotypes, the lowest seroreactivity was found against B. quintana (11.2%) and the highest, against B. v. berkhoffii genotype III (56%). A total of 27% of 89 individuals were not seroreactive to any test antigen. Bartonella spp. IFA seroreactivity was not associated with any clinical sign or symptom. DNA from Bartonella spp., including B. henselae (n = 2), B. v. berkhoffii genotypes I (n = 1) and III (n = 2), and B. quintana (n = 2) was detected in 7/89 veterinary personnel. PCR and DNA sequencing findings were not associated with clinical signs or symptoms. No co-infections were observed. One of the two B. henselae PCR-positive individuals was IFA seronegative to all tested antigens whereas the other one was not B. henselae seroreactive. The remaining PCR-positive individuals were seroreactive to multiple Bartonella spp. antigens. High serological and molecular prevalences of exposure to, or infection with, Bartonella spp. were found in companion animal veterinary personnel from Spain. More studies using BAPGM enrichment blood culture and PCR are needed to clarify the finding of Bartonella PCR-positive individuals lacking clinical symptoms. DA - 2017/11/7/ PY - 2017/11/7/ DO - 10.1186/s13071-017-2483-z VL - 10 IS - 1 SP - J2 - Parasites Vectors LA - en OP - SN - 1756-3305 UR - http://dx.doi.org/10.1186/s13071-017-2483-z DB - Crossref KW - Bartonella alphaproteobacteria growth medium KW - Bartonella henselae KW - Bartonella quintana KW - Bartonella vinsonii berkhoffii KW - Bartonella koehlerae KW - Veterinary personnel KW - Spain ER - TY - JOUR TI - Population Structure of Pseudocercospora fijiensis in Costa Rica Reveals Shared Haplotype Diversity with Southeast Asian Populations AU - Saville, Amanda AU - Charles, Melodi AU - Chavan, Suchitra AU - Muñoz, Miguel AU - Gómez-Alpizar, Luis AU - Ristaino, Jean Beagle T2 - Phytopathology AB - Pseudocercospora fijiensis is the causal pathogen of black Sigatoka, a devastating disease of banana that can cause 20 to 80% yield loss in the absence of fungicides in banana crops. The genetic structure of populations of P. fijiensis in Costa Rica was examined and compared with Honduran and global populations to better understand migration patterns and inform management strategies. In total, 118 isolates of P. fijiensis collected from Costa Rica and Honduras from 2010 to 2014 were analyzed using multilocus genotyping of six loci and compared with a previously published global dataset of populations of P. fijiensis. The Costa Rican and Honduran populations shared haplotype diversity with haplotypes from Southeast Asia, Oceania, and the Americas but not Africa for all but one of the six loci studied. Gene flow and shared haplotype diversity was found in Honduran and Costa Rican populations of the pathogen. The data indicate that the haplotypic diversity observed in Costa Rican populations of P. fijiensis is derived from dispersal from initial outbreak sources in Honduras and admixtures between genetically differentiated sources from Southeast Asia, Oceania, and the Americas. DA - 2017/12// PY - 2017/12// DO - 10.1094/phyto-02-17-0045-r VL - 107 IS - 12 SP - 1541–1548 SN - 0031-949X UR - http://dx.doi.org/10.1094/PHYTO-02-17-0045-R ER - TY - JOUR TI - A VIGS screen identifies immunity in the Arabidopsis Pla-1 accession to viruses in two different genera of the Geminiviridae AU - Reyes, Maria Ines AU - Flores-Vergara, Miguel A. AU - Guerra-Peraza, Orlene AU - Rajabu, Cyprian AU - Desai, Jigar AU - Hiromoto-Ruiz, Yokiko H. AU - Ndunguru, Joseph AU - Hanley-Bowdoin, Linda AU - Kjemtrup, Susanne AU - Ascencio-Ibanez, Jose T. AU - Robertson, Dominique T2 - PLANT JOURNAL AB - Summary Geminiviruses are DNA viruses that cause severe crop losses in different parts of the world, and there is a need for genetic sources of resistance to help combat them. Arabidopsis has been used as a source for virus‐resistant genes that derive from alterations in essential host factors. We used a virus‐induced gene silencing ( VIGS ) vector derived from the geminivirus Cabbage leaf curl virus (Ca LC uV) to assess natural variation in virus–host interactions in 190 Arabidopsis accessions. Silencing of CH ‐42 , encoding a protein needed to make chlorophyll, was used as a visible marker to discriminate asymptomatic accessions from those showing resistance. There was a wide range in symptom severity and extent of silencing in different accessions, but two correlations could be made. Lines with severe symptoms uniformly lacked extensive VIGS , and lines that showed attenuated symptoms over time (recovery) showed a concomitant increase in the extent of VIGS . One accession, Pla‐1, lacked both symptoms and silencing, and was immune to wild‐type infectious clones corresponding to Ca LC uV or Beet curly top virus ( BCTV ), which are classified in different genera in the Geminiviridae. It also showed resistance to the agronomically important Tomato yellow leaf curl virus ( TYLCV ). Quantitative trait locus mapping of a Pla‐1 X Col‐0 F 2 population was used to detect a major peak on chromosome 1, which is designated gip‐1 ( geminivirus immunity Pla‐1‐1 ). The recessive nature of resistance to Ca LC uV and the lack of obvious candidate genes near the gip‐1 locus suggest that a novel resistance gene(s) confers immunity. DA - 2017/12// PY - 2017/12// DO - 10.1111/tpj.13716 VL - 92 IS - 5 SP - 796-807 SN - 1365-313X KW - Arabidopsis KW - BCTV KW - CaLCuV KW - geminivirus KW - gip-1 KW - immunity KW - Pla-1 KW - VIGS ER - TY - JOUR TI - Repliscan: a tool for classifying replication timing regions AU - Zynda, Gregory J. AU - Song, Jawon AU - Concia, Lorenzo AU - Wear, Emily E. AU - Hanley-Bowdoin, Linda AU - Thompson, William F. AU - Vaughn, Matthew W. T2 - BMC BIOINFORMATICS AB - Replication timing experiments that use label incorporation and high throughput sequencing produce peaked data similar to ChIP-Seq experiments. However, the differences in experimental design, coverage density, and possible results make traditional ChIP-Seq analysis methods inappropriate for use with replication timing.To accurately detect and classify regions of replication across the genome, we present Repliscan. Repliscan robustly normalizes, automatically removes outlying and uninformative data points, and classifies Repli-seq signals into discrete combinations of replication signatures. The quality control steps and self-fitting methods make Repliscan generally applicable and more robust than previous methods that classify regions based on thresholds.Repliscan is simple and effective to use on organisms with different genome sizes. Even with analysis window sizes as small as 1 kilobase, reliable profiles can be generated with as little as 2.4x coverage. DA - 2017/8/7/ PY - 2017/8/7/ DO - 10.1186/s12859-017-1774-x VL - 18 SP - 1-14 SN - 1471-2105 UR - http://europepmc.org/abstract/med/28784090 KW - DNA replication KW - Repli-seq KW - Classification ER - TY - JOUR TI - Neurological and immunological dysfunction in two patients with Bartonella henselae bacteremia AU - Kaufman, David L. AU - Kogelnik, Andreas M. AU - Mozayeni, Robert B. AU - Cherry, Natalie A. AU - Breitschwerdt, Edward B. T2 - Clinical Case Reports AB - Recently, BAPGM enrichment culture has documented Bartonella bacteremia in previously healthy, “nonimmunocompromised” patients following arthropod exposures. Neurobartonellosis should be among the differential diagnoses for patients with persistent or recurrent neurological symptoms of undetermined etiology. Microbiological and immunological testing should be concurrently pursued to determine whether defective immune function accompanies Bartonella bacteremia. A 49-year-old female veterinarian from California was previously healthy working 60 h a week managing a cat shelter with 500 cats. These factors may have predisposed acquisition of B. henselae and failure to immunologically eliminate the infection. In October 2011, she experienced an extensive infestation of mange mites (Sarcoptes scabiei or Notoedres cati) after handling an infested cat. One month later, she experienced an episode of confusion and severe headache. There was no prior history of headaches or migraines. That evening, she again became confused, experienced a 15-min episode of total aphasia, witnessed by her husband, and had a headache. All symptoms resolved after 3 h. Over the next few months, she continued to have headaches, episodic confusion, and aphasia. Because an aura preceded symptoms, she would stop performing surgery, sit down, or stop driving. Initially, these episodes occurred one to two times a month; however, over the next 4 years, all symptoms increased in frequency and duration, occurring 20–25 days a month and lasting minutes to hours. She subsequently developed seizures and balance and disequilibrium problems causing gait impairment and falls. She was examined by multiple physicians. An extensive neurological workup included an MRI performed in August 2013; all results were negative except for electroencephalographic “left temporal lobe slowing.” After treatment with the anticonvulsant levetiracetam, she reported a 3-week period of mental clarity and felt normal. Subsequently, symptoms recurred and she did not respond to dose adjustment or additional medications. Her illness progressively worsened, and she stopped working 4 years after symptom onset. Based on her clinical history, high arthropod and feral cat exposure risk, ambiguous neurological test results, and failure to improve with medical therapy, a search for an infectious cause was initiated. Initial serology results (Quest Diagnostics, Inc.) were negative for Borrelia burgdorferi, Anaplasma phagocytophilum, Ehrlichia chaffeensis, Babesia, Bartonella, Toxoplasma, and Cryptococcus. In December 2014, a repeat MRI revealed left frontal convexity falx calcifications. A SPECT scan documented heterogeneous decreased uptake bilaterally in the frontal and anterior temporal lobes with greater decrease involving the left hemisphere. Cerebrospinal fluid analyses were within laboratory reference ranges. Due to progressive illness and lack of a diagnosis, immunological testing was performed. There were decreased total IgG and deficiencies in subclasses 1, 2, and 3. CD3, CD4, and CD8 absolute cell counts were abnormally low, and a natural killer cell (NK) function assay was profoundly low (Table 1). Because immunological test results were abnormal, additional infectious disease testing was pursued. Given her daily work with cats, frequent exposure to fleas, and historical mite infestation, Bartonella spp. bacteremia remained a diagnostic consideration. EDTA-anticoagulated blood specimens were screened for Bartonella using the Bartonella alpha-proteobacteria growth medium (BAPGM) enrichment culture/PCR diagnostic platform. (Bartonella enrichment PCR™ or ePCR™ (Galaxy Diagnostics, Inc., Research Triangle Park, NC)) Serological analysis of Bartonella henselae and Bartonella quintana was performed using indirect fluorescent antibody (IFA) testing (Galaxy Diagnostics, Inc., Research Triangle Park, NC). Bartonella henselae DNA was amplified and sequenced from a 21-day BAPGM enrichment blood culture. Serum was not reactive to B. henselae or B. quintana antigens at 1:16 or 1:32 screening dilutions. Genus PCR assays (Galaxy Diagnostics, Inc., Research Triangle Park, NC) for Anaplasma, Babesia, Ehrlichia, and Rickettsia spp. were negative. Based on these results, she was treated with clarithromycin, rifampin, and intravenous gamma globulin every 3 weeks. By week six of antibiotic/IVIG administration, her headaches slowly improved, and she returned to work. After 9 months of antibiotics, she was B. henselae and B. quintana seroreactive at titers of 1:128 and 1:64, respectively. Three BAPGM enrichment blood cultures collected on alternate days were PCR negative. During the subsequent 9-month follow-up, she has experienced no seizures. An MRI, repeated 14 months after the December 2014 MRI study, and 11 months after beginning antibiotics, was unchanged. Her previously abnormal lymphocyte subsets normalized, although NK function remained depressed at 10. A 69-year-old previously healthy woman hiking in New York State during November 2013 became acutely ill 3 weeks after returning to California. She reported flu-like symptoms including headaches, nausea, vertigo with disequilibrium, and fatigue but no fever or pain. She developed a bulls-eye rash on her right infraclavicular fossa, consistent with erythema chronicum migrans. Lyme disease was diagnosed by another physician, who initiated doxycycline treatment for 2 weeks. She continued to have symptoms and was examined by the primary author 10 days after starting doxycycline (Open Medicine Institute, Mountain View, CA). Laboratory testing results for Borrelia burgdorferi, Anaplasma phagocytophilum, Ehrlichia chaffeensis, Babesia, and Bartonella antibodies (Quest Diagnostics, Inc.) were negative. Doxycycline was continued for 4 weeks resulting in a transient decrease in symptoms. Headaches, vertigo, disequilibrium, and nausea recurred 2 weeks after completing antibiotics. Treatment with doxycycline and azithromycin initiated on January 2014 resulted in symptomatic improvement; however, antibiotic administration ceased after 2 weeks due to a photosensitivity reaction while visiting Florida. Symptoms returned after 2 weeks, only to resolve again with amoxicillin and azithromycin administration. Within 5 days, she developed severe diarrhea, antibiotics were stopped, and within 3 days, her headache and vertigo returned. A neurology workup in August 2014, including MRI, MRA, and cerebral angiogram, detected no abnormalities. In October 2014, she was treated with minocycline and metronidazole for headaches, vertigo, and disequilibrium. After 5 weeks, there was symptomatic improvement that continued through 8 weeks, when antibiotics were stopped. Six weeks later, all symptoms recurred. In April 2015, immunological testing documented an IgG subclass 3 deficiency; low absolute CD3, CD8, and CD19 cell counts; and decreased NK cell function (Table 1). Repeat serological testing (Quest Diagnostics, Inc.) for tickborne diseases remained negative. Bartonella henselae DNA was amplified and sequenced from a 21-day BAPGM enrichment blood culture (Bartonella enrichment PCR™ or ePCR™ (Galaxy Diagnostics, Inc., Research Triangle Park, NC)). Serum was not B. henselae or B. quintana seroreactive at 1:16 or 1:32 screening dilutions. Genus PCR assays (Galaxy Diagnostics, Inc., Research Triangle Park, NC) for Anaplasama, Babesia, Ehrlichia, and Rickettsia spp. were negative. After being treated with clarithromycin and rifampin for 5 months, headaches and vertigo were almost completely resolved and she remained B. henselae and B. quintana IFA seronegative, although lymphocyte subsets and NK function remained abnormal. She remained B. henselae or B. quintana seronegative, and three BAPGM enrichment blood cultures collected on alternate days were PCR negative. Two important clinical observations evolved out of the microbiological, immunological, and therapeutic findings associated with the medical management of these two patients. First, persistent or recurrent neurological symptoms of undetermined etiology in patients with historical vector exposures should prompt testing for bartonellosis. Historically, B. henselae infections in immunocompetent individuals have been associated with self-limiting cat scratch disease, whereas recent research supports persistent and potentially relapsing bacteremia 1-3. As previously reported 2, 3, both of these B. henselae bacteremic patients experienced headaches and disequilibrium. Patient 1 also experienced seizures, episodic confusion, and aphasia, which resolved completely following antibiotic therapy, despite persistence of the MRI abnormalities 1. Secondly, immunological testing should be concurrently pursued to determine whether defective immune function accompanies neurological symptoms. Both patients had immunological abnormalities, including suppression of NK function, despite lacking a prior medical history indicative of immunodeficiency. The extent to which persistent B. henselae bacteremia may have induced immunocompromise or whether a chronic latent infection resulted in bacterial reactivation is unknown. It is important to note that after 9 months of treatment, the CD3, CD4, and CD19 deficiencies in Patient 1 resolved, while Patient 2 remained immunologically impaired after 5 months of therapy. There remains a substantial need for sequential electroencephalographic, MRI, immunological, and bacteriological patient data to guide physician decision making in patients with longstanding B. henselae bacteremia. Using a previously validated diagnostic approach 3, 4, B. henselae bacteremia was confirmed in both patients by BAPGM enrichment blood culture, PCR amplification, and DNA sequence confirmation. Importantly, PCR did not amplify B. henselae DNA from patient's blood, serum, 8-day, or 14-day BAPGM enrichment blood cultures, supporting the need for prolonged bacterial incubation times to obtain PCR confirmation for some B. henselae bacteremic patients. Consistent with previous studies3, 5 in which a subset of patients with persistent bacteremia were not IFA seroreactive to a panel of Bartonella sp. antigens, neither patient was initially B. henselae or B. quintana IFA seroreactive, whereas antibody reactivity was documented after antibiotic treatment in Patient 1, potentially due to enhanced immunological recognition of antigenic epitopes. Based upon these patients and previously published studies 3, 5, enrichment blood culture and PCR should be used in conjunction with Bartonella sp. serological analysis when attempting to confirm bacteremic infection with a Bartonella sp. Vector transmission of B. henselae was the suspected source of infection for both patients. The veterinarian had ongoing vector (flea, mite, and potentially ticks) and animal exposure, which are occupational risks for animal health workers 3, 5. The cat flea (Ctenocephalides felis) transmits B. henselae among cats that develop a relapsing bacteremia and remain persistently infected reservoir hosts for months to years 6. Although there is no evidence that cat-associated mites (S. scabiei or N. cati) are vector competent for the transmission of Bartonella species, rat mite (Ornithonyssus bacoti) and pigeon mite (Dermanyssus spp.) transmissions of B. henselae and B. quintana, respectively, have been suspected 7, 8. Due to an acute-onset illness and presumed tick attachment in a Lyme-endemic region with the subsequent development of erythema chronicum migrans, Lyme disease was initially suspected in Patient 2. Rapid treatment with doxycycline may have prevented serodiagnostic confirmation of B. burgdorferi transmission, whereas testing for other tickborne pathogens was negative. Although tick transmission of B. henselae has not been proven, organism-specific DNA has been PCR-amplified and sequenced from Ixodes sp. ticks 9, vector competence for Bartonella transmission has been demonstrated in a rodent model 10, and French investigators have recently documented Bartonella spp. bacteremia in patients following tick exposures 11. Historically, systemic bartonellosis has been reported in immunocompromised patients, such as those with HIV/AIDS and transplant recipients. Recently, infection with Bartonella spp. has been reported in healthy asymptomatic Brazilian blood donor candidates12 and in previously immunocompetent patients with chronic neurological or rheumatologic symptoms 2, 3, 5. Because Bartonella spp. can infect erythrocytes, endothelial cells, and various macrophage-type cells, including brain-derived dendritic cells in vitro, the spectrum of neurological symptoms attributable to bartonellosis appear to be extremely diverse among patients 1, 2. Physicians should be aware of the rapidly increasing number of Bartonella spp., the large number of proven and suspected arthropod vectors, and the large number of reservoir hosts, all of which are collectively contributing to the enhanced recognition of neurobartonellosis as a medically important emerging infectious disease. We would like to thank Rowena Moss for her efforts in patient testing and editorial assistance. DK: principal investigator and physician of patients presented in this publication – analyzed clinical data and prepared the manuscript. AMK: provided medical consultation and received funding resources. RBM: provided medical consultation. NAC: involved in laboratory data collection and analysis and prepared the manuscript. EBB, MD: principal investigator – is involved in data analysis and manuscript preparation. In conjunction with Dr. Sushama Sontakke and North Carolina State University, Edward B. Breitschwerdt, DVM, holds U.S. Patent No. 7,115,385; Media and Methods for cultivation of microorganisms, which was issued on 3 October 2006. He is a co-founder, shareholder, and chief scientific officer for Galaxy Diagnostics, a company that provides advanced diagnostic testing for the detection of Bartonella species infections. Robert B. Mozayeni, MD, is chief medical officer, and Natalie Cherry, PhD, is the laboratory supervisor and research analyst for Galaxy Diagnostics. The remaining authors have no competing interests. DA - 2017/4/26/ PY - 2017/4/26/ DO - 10.1002/ccr3.977 VL - 5 IS - 6 SP - 931-935 J2 - Clin Case Rep LA - en OP - SN - 2050-0904 UR - http://dx.doi.org/10.1002/ccr3.977 DB - Crossref KW - Bartonella KW - immune dysfunction KW - Infection KW - neurological symptoms KW - vectorborne ER - TY - JOUR TI - Genetic Architecture of Natural Variation Underlying Adult Foraging Behavior That Is Essential for Survival of Drosophila melanogaster AU - Lee, Yuh Chwen G. AU - Yang, Qian AU - Chi, Wanhao AU - Turkson, Susie A. AU - Du, Wei A. AU - Kemkemer, Claus AU - Zeng, Zhao-Bang AU - Long, Manyuan AU - Zhuang, Xiaoxi T2 - GENOME BIOLOGY AND EVOLUTION AB - Foraging behavior is critical for the fitness of individuals. However, the genetic basis of variation in foraging behavior and the evolutionary forces underlying such natural variation have rarely been investigated. We developed a systematic approach to assay the variation in survival rate in a foraging environment for adult flies derived from a wild Drosophila melanogaster population. Despite being such an essential trait, there is substantial variation of foraging behavior among D. melanogaster strains. Importantly, we provided the first evaluation of the potential caveats of using inbred Drosophila strains to perform genome-wide association studies on life-history traits, and concluded that inbreeding depression is unlikely a major contributor for the observed large variation in adult foraging behavior. We found that adult foraging behavior has a strong genetic component and, unlike larval foraging behavior, depends on multiple loci. Identified candidate genes are enriched in those with high expression in adult heads and, demonstrated by expression knock down assay, are involved in maintaining normal functions of the nervous system. Our study not only identified candidate genes for foraging behavior that is relevant to individual fitness, but also shed light on the initial stage underlying the evolution of the behavior. DA - 2017/5// PY - 2017/5// DO - 10.1093/gbe/evx089 VL - 9 IS - 5 SP - 1357-1369 SN - 1759-6653 KW - genome-wide association study KW - foraging behavior KW - Drosophila melanogaster KW - inbreeding depression ER - TY - JOUR TI - Characterization of Hydroporphyrins Covalently Attached to Si(100) AU - Jiao, Jieying AU - Yu, Miao AU - Holten, Dewey AU - Lindsey, Jonathan S. AU - Bocian, David F. T2 - JOURNAL OF PORPHYRINS AND PHTHALOCYANINES AB - Attachment of synthetic analogs of natural tetrapyrroles to electroactive surfaces enables physicochemical interrogation and may provide material for use in catalysis, diagnostics, and energy conversion. Six synthetic zinc chlorins and one free base bacteriochlorin, tailored analogs of chlorophyll and bacteriochlorophyll, respectively, have been attached to Si(100) via a high-temperature (400°C) baking method. The hydroporphyrins bear diverse functional groups that enable surface attachment (vinyl, acetyl, triisopropylsilylethynyl, pentafluorophenyl, and hydroxymethylphenyl) and a geminal dimethyl group in each reduced ring for stabilization toward adventitious dehydrogenation. The films were examined by cyclic voltammetry, FTIR spectroscopy, X-ray photoelectron spectroscopy, and ellipsometry. Monofunctionalized and difunctionalized hydroporphyrins gave monolayer and multilayer films, respectively, indicating robustness of the hydroporphyrin molecules, but in each case the film was more heterogeneous than observed with comparable porphyrins. The data suggest that some amount of unattached molecules remain intercalated with surface-attached molecules. Additional molecular designs will need to be examined to develop a deep understanding of the structure-activity relationship for formation of homogeneous monolayers and multilayers of synthetic hydroporphyrins. DA - 2017/// PY - 2017/// DO - 10.1142/s1088424617500547 VL - 21 IS - 7-8 SP - 453-464 SN - 1099-1409 KW - hydroporphyrins KW - Si(100) KW - cyclic voltammetry KW - FTIR spectroscopy KW - X-ray photoelectron spectroscopy KW - ellipsometry ER - TY - JOUR TI - Vector-borne and zoonotic diseases of dogs in North-west New South Wales and the Northern Territory, Australia AU - Shapiro, Amanda J. AU - Brown, Graeme AU - Norris, Jacqueline M. AU - Bosward, Katrina L. AU - Marriot, Debbie J. AU - Balakrishnan, Nandhakumar AU - Breitschwerdt, Edward B. AU - Malik, Richard T2 - BMC Veterinary Research AB - Vector-borne diseases of dogs in Australian Aboriginal communities are relatively unexplored. These dogs represent a unique group with variable ecto- and endo-parasitic burdens, nutritional stresses and a general lack of veterinary intervention. We investigated haemoprotozoal and bacterial pathogen prevalences in relation to erythrocyte and platelet numbers in dogs from North-West New South Wales (N-W NSW) and the Northern Territory (NT; Central Australia).Real-time PCR (qPCR) amplification of Anaplasma platys, Babesia vogeli, Mycoplasma haemocanis, Candidatus Mycoplasma haematoparvum and Bartonella spp., serological screening for Coxiella burnetii, and Bartonella spp. and haematological analyses were performed on dogs from the two cohorts (96 dogs in total). Brucella suis serology was determined additionally for the N-W NSW cohort.Anaplasma platys (n = 26 dogs), Babesia vogeli (n = 7), Candidatus Mycoplasma haematoparvum (n = 10 dogs), and Mycoplasma haemocanis (n = 14) were detected in the sample population (n = 96) using qPCR. There were significant associations between (i) A. platys and anaemia (OR 8.7, CI 2.4-31.7; P < 0.001), thrombocytopenia (OR 12.1, CI 3.4-43.2; P < 0.001) and breed (OR 16.1, CI 2.1-121.5; P = 0.007), and (ii) between B. vogeli and anaemia (OR 11.8, CI 2.3-61.6; P = 0.003). Neither protozoal nor bacterial DNA loads, estimated using qPCR, were positively correlated with anaemia or thrombocytopenia. Haemotropic mycoplasmas were not associated with any haematologic abnormality. Four dogs from the NT were seropositive for Coxiella burnetii, while no dogs were seropositive for Brucella suis or to a panel of Bartonella spp. antigens. Despite directed efforts, Bartonella DNA was not detected in blood from any of the cohorts studied. A sample of dogs from the NT recruited specifically for Bartonella α-proteobacteria growth medium enrichment blood culture were also Bartonella PCR negative.Vector-borne pathogens occur in dogs free ranging near Aboriginal communities, with higher detection rates in NT than N-W NSW. The preponderant haematologic abnormalities were anaemia and thrombocytopenia, likely attributable to A. platys and B. vogeli infections, but also probably affected by nutritional, parasitic, lactational and environmental stressors. The absence of Bartonella spp. is of importance to the Australian setting, and work needs to be extended to tropical coastal communities where fleas are present as well as ticks. Dogs living in and around Aboriginal communities may provide valuable sentinel information on disease infection status of human public health significance. DA - 2017/8/15/ PY - 2017/8/15/ DO - 10.1186/s12917-017-1169-2 VL - 13 IS - 1 SP - J2 - BMC Vet Res LA - en OP - SN - 1746-6148 UR - http://dx.doi.org/10.1186/s12917-017-1169-2 DB - Crossref KW - Dog KW - Babesia spp. KW - Anaplasma spp. KW - Haemotropic mycoplasmas KW - Coxiella burnetii KW - Bartonella spp. KW - Brucella spp. ER - TY - JOUR TI - Prevalence of Vector-Borne Pathogens in Southern California Dogs With Clinical and Laboratory Abnormalities Consistent With Immune-Mediated Disease AU - Kidd, L. AU - Qurollo, B. AU - Lappin, M. AU - Richter, K. AU - Hart, J.R. AU - Hill, S. AU - Osmond, C. AU - Breitschwerdt, E.B. T2 - Journal of Veterinary Internal Medicine AB - Background Studies investigating the prevalence of vector‐borne pathogens in southern California dogs are limited. Occult infections might be misdiagnosed as idiopathic immune‐mediated disease. Hypothesis/Objectives (1) To determine the prevalence of vector‐borne pathogens in southern California dogs with compatible clinical findings using PCR and serologic panels and (2) to determine whether testing convalescent samples and repeating PCR on acute samples using the same and different gene targets enhance detection. Animals Forty‐two client‐owned dogs with clinical signs of vector‐borne disease presenting to specialty practices in San Diego County. Methods Combined prospective and retrospective observational study. Forty‐two acute and 27 convalescent samples were collected. Acute samples were prospectively tested for antibodies to Rickettsia, Ehrlichia, Bartonella, Babesia, Borrelia, and Anaplasma species . PCR targeting Ehrlichia , Babesia , Anaplasma , hemotropic Mycoplasma, and Bartonella species was also performed. Retrospectively, convalescent samples were tested for the same organisms using serology, and for Ehrlichia , Babesia , Anaplasma , and Bartonella species using PCR. Acute samples were retested using PCR targeting Ehrlichia and Babesia species. Results Evidence of exposure to or infection with a vector‐borne pathogen was detected in 33% (14/42) of dogs. Ehrlichia and Babesia species were most common; each was identified in 5 dogs. Convalescent serologic testing, repeating PCR, and using novel PCR gene targets increased detection by 30%. Conclusions and Clinical Importance Repeated testing using serology and PCR enhances detection of infection by vector‐borne pathogens in dogs with clinical signs of immune‐mediated disease. Larger prevalence studies of emerging vector‐borne pathogens in southern California dogs are warranted. DA - 2017/5/30/ PY - 2017/5/30/ DO - 10.1111/jvim.14735 VL - 31 IS - 4 SP - 1081-1090 J2 - J Vet Intern Med LA - en OP - SN - 0891-6640 UR - http://dx.doi.org/10.1111/jvim.14735 DB - Crossref KW - Anaplasmosis KW - Babesiosis KW - Ehrlichiosis KW - Flea KW - Immune-mediated KW - Rickettsioses KW - Tick ER - TY - JOUR TI - Culture, PCR, DNA sequencing, and second harmonic generation (SHG) visualization of Bartonella henselae from a surgically excised human femoral head AU - Ericson, M. AU - Balakrishnan, N. AU - Mozayeni, B. R. AU - Woods, C. W. AU - Dencklau, J. AU - Kelly, S. AU - Breitschwerdt, E. B. T2 - CLINICAL RHEUMATOLOGY DA - 2017/7// PY - 2017/7// DO - 10.1007/s10067-016-3524-2 VL - 36 IS - 7 SP - 1669-1675 SN - 1434-9949 ER - TY - JOUR TI - Synthesis and spectral properties of meso-arylbacteriochlorins, including insights into essential motifs of their hydrodipyrrin precursors AU - Reddy, M. N. AU - Zhang, S. F. AU - Kim, H. J. AU - Mass, O. AU - Taniguchi, M. AU - Lindsey, J. S. T2 - Molecules DA - 2017/// PY - 2017/// VL - 22 IS - 4 ER - TY - JOUR TI - Seroprevalence of Bartonella species, Coxiella burnetii and Toxoplasma gondii among patients with hematological malignancies: A pilot study in Romania AU - Messinger, C. J. AU - Gurzau, E. S. AU - Breitschwerdt, E. B. AU - Tomuleasa, C. I. AU - Trufan, S. J. AU - Flonta, M. M. AU - Maggi, R. G. AU - Berindan-Neagoe, I. AU - Rabinowitz, P. M. T2 - Zoonoses and Public Health AB - Summary Patients receiving immunosuppressive cancer treatments in settings where there is a high degree of human–animal interaction may be at increased risk for opportunistic zoonotic infections or reactivation of latent infections. We sought to determine the seroprevalence of selected zoonotic pathogens among patients diagnosed with haematologic malignancies and undergoing chemotherapeutic treatments in Romania, where much of the general population lives and/or works in contact with livestock. A convenience sample of 51 patients with haematologic cancer undergoing chemotherapy at a referral clinic in Cluj‐Napoca, Romania, was surveyed regarding animal exposures. Blood samples were obtained and tested for evidence of infection with Bartonella species, Coxiella burnetii and Toxoplasma gondii , which are important opportunistic zoonotic agents in immunocompromised individuals. 58.8% of participants reported living or working on a farm, and living or working on a farm was associated with contact with livestock and other animals. 37.5% of participants were IgG seroreactive against one or more of five Bartonella antigens, and seroreactivity was statistically associated with living on farms. Farm dwellers were 3.6 times more likely to test IgG seroreactive to Bartonella antibodies than non‐farm dwellers. 47.1% of the participants tested T. gondii IgG positive and 13.7% tested C. burnetii IgG positive, indicating past or latent infection. C. burnetii IgM antibodies were detected in four participants (7.8%), indicating possible recent infection. These results indicate that a large proportion of patients with haematologic cancer in Romania may be at risk for zoonotic infections or for reactivation of latent zoonotic infections, particularly with respect to Bartonella species. Special attention should be paid to cancer patients' exposure to livestock and companion animals in areas where much of the population lives in rural settings. DA - 2017/3/22/ PY - 2017/3/22/ DO - 10.1111/zph.12353 VL - 64 IS - 6 SP - 485-490 J2 - Zoonoses Public Health LA - en OP - SN - 1863-1959 UR - http://dx.doi.org/10.1111/zph.12353 DB - Crossref KW - Bartonella KW - Coxiella burnetii KW - immunocompromised patients KW - livestock KW - seroprevalence KW - Toxoplasma KW - zoonoses ER - TY - JOUR TI - Intracoronary allogeneic cardiosphere-derived stem cells are safe for use in dogs with dilated cardiomyopathy AU - Hensley, Michael Taylor AU - Tang, Junnan AU - Woodruff, Kathleen AU - Defrancesco, Teresa AU - Tou, Sandra AU - Williams, Christina M. AU - Breen, Mathew AU - Meurs, Kathryn AU - Keene, Bruce AU - Cheng, Ke T2 - JOURNAL OF CELLULAR AND MOLECULAR MEDICINE AB - Cardiosphere-derived cells (CDCs) have been shown to reduce scar size and increase viable myocardium in human patients with mild/moderate myocardial infarction. Studies in rodent models suggest that CDC therapy may confer therapeutic benefits in patients with non-ischaemic dilated cardiomyopathy (DCM). We sought to determine the safety and efficacy of allogeneic CDC in a large animal (canine) model of spontaneous DCM. Canine CDCs (cCDCs) were grown from a donor dog heart. Similar to human CDCs, cCDCs express CD105 and are slightly positive for c-kit and CD90. Thirty million of allogeneic cCDCs was infused into the coronary vessels of Doberman pinscher dogs with spontaneous DCM. Adverse events were closely monitored, and cardiac functions were measured by echocardiography. No adverse events occurred during and after cell infusion. Histology on dog hearts (after natural death) revealed no sign of immune rejection from the transplanted cells. DA - 2017/8// PY - 2017/8// DO - 10.1111/jcmm.13077 VL - 21 IS - 8 SP - 1503-1512 SN - 1582-4934 KW - cardiosphere-derived cells KW - dogs KW - dilated cardiomyopathy KW - stem cell therapy KW - allogeneic ER - TY - JOUR TI - Integrated immunohistochemical and DNA copy number profiling analysis provides insight into the molecular pathogenesis of canine follicular lymphoma AU - Thomas, R. AU - Demeter, Z. AU - Kennedy, K. A. AU - Borst, L. AU - Singh, K. AU - Valli, V. E. AU - Le Boedec, K. AU - Breen, M. T2 - VETERINARY AND COMPARATIVE ONCOLOGY AB - Follicular lymphomas (FLs) typically exhibit a chromosome translocation that induces constitutive expression of the anti-apoptotic bcl2 protein and accumulation of additional molecular defects. This rearrangement offers a promising therapeutic target, but its nature as a fundamental driver of FL pathogenesis remains unclear as 15% of cases lack the translocation. We performed an integrated immunohistochemical and genomic investigation of 10 naturally occurring FL cases from domestic dogs, showing that, as with human tumours, they exhibit marked heterogeneity in the frequency and intensity of bcl2 protein expression. Genomic copy number aberrations were infrequent and broadly consistent with those of other canine B-cell lymphoma subtypes. None of the canine FL specimens exhibited a rearrangement consistent with the hallmark translocation of human FL, despite their remarkable histomorphologic similarity. Parallel exploration of canine and human cases may reveal alternative tumour-initiating mechanisms other than BCL2 disruption, yielding a more complete definition of the molecular pathogenesis of FL. DA - 2017/9// PY - 2017/9// DO - 10.1111/vco.12227 VL - 15 IS - 3 SP - 852-867 SN - 1476-5829 KW - BCL2 KW - canine KW - comparative genomic hybridization KW - fluorescence in situ hybridization KW - follicular lymphoma KW - immunohistochemistry ER - TY - JOUR TI - Association of breed and histopathological grade in canine mast cell tumours AU - Mochizuki, H. AU - Motsinger-Reif, A. AU - Bettini, C. AU - Moroff, S. AU - Breen, M. T2 - VETERINARY AND COMPARATIVE ONCOLOGY AB - Abstract The aim of this study was to evaluate the relationship between breed and the histopathological grade of canine mast cell tumours ( MCTs ). A retrospective survey of pathology data of 9375 histopathologically confirmed diagnoses of cutaneous MCTs in the US was evaluated in the context of breed prevalence in over two million registered purebred dogs. Association of histopathological grade with breed, age, sex and spay/neuter status was assessed. The data indicate that the proportion of high‐grade tumours increases with advancing age, and that male and intact dogs have increased odds of developing high‐grade tumours. A significant difference in the proportion of high‐grade tumours between breeds was detected. The Pug was at significantly increased risk of developing low/intermediate‐grade tumours, but not high‐grade tumours, resulting in preponderance of less aggressive MCTs in this breed. The results of this study suggest a genetic association for the development of high‐grade MCTs . DA - 2017/9// PY - 2017/9// DO - 10.1111/vco.12225 VL - 15 IS - 3 SP - 829-839 SN - 1476-5829 KW - Boxer KW - dog KW - epidemiology KW - genetics KW - MCT KW - Pug ER - TY - JOUR TI - Networks Underpinning Symbiosis Revealed Through Cross-Species eQTL Mapping AU - Guo, Yuelong AU - Fudali, Sylwia AU - Gimeno, Jacinta AU - DiGennaro, Peter AU - Chang, Stella AU - Williamson, Valerie M. AU - Bird, David Mck. AU - Nielsen, Dahlia M. T2 - GENETICS AB - Abstract Interactions between species are pervasive among plants, animals, and microbes, and identifying the molecular signals involved is an active area of research.. Organisms engage in extensive cross-species molecular dialog, yet the underlying molecular actors are known for only a few interactions. Many techniques have been designed to uncover genes involved in signaling between organisms. Typically, these focus on only one of the partners. We developed an expression quantitative trait locus (eQTL) mapping-based approach to identify cause-and-effect relationships between genes from two partners engaged in an interspecific interaction. We demonstrated the approach by assaying expression of 98 isogenic plants (Medicago truncatula), each inoculated with a genetically distinct line of the diploid parasitic nematode Meloidogyne hapla. With this design, systematic differences in gene expression across host plants could be mapped to genetic polymorphisms of their infecting parasites. The effects of parasite genotypes on plant gene expression were often substantial, with up to 90-fold (P = 3.2 × 10−52) changes in expression levels caused by individual parasite loci. Mapped loci included a number of pleiotropic sites, including one 87-kb parasite locus that modulated expression of &gt;60 host genes. The 213 host genes identified were substantially enriched for transcription factors. We distilled higher-order connections between polymorphisms and genes from both species via network inference. To replicate our results and test whether effects were conserved across a broader host range, we performed a confirmatory experiment using M. hapla-infected tomato. This revealed that homologous genes were similarly affected. Finally, to validate the broader utility of cross-species eQTL mapping, we applied the strategy to data from a Salmonella infection study, successfully identifying polymorphisms in the human genome affecting bacterial expression. DA - 2017/8// PY - 2017/8// DO - 10.1534/genetics.117.202531 VL - 206 IS - 4 SP - 2175-2184 SN - 1943-2631 UR - http://www.scopus.com/inward/record.url?eid=2-s2.0-85027059202&partnerID=MN8TOARS KW - transspecies KW - trans-eQTL KW - host-pathogen interaction KW - symbiosis KW - RNA-Seq ER - TY - JOUR TI - Evaluation of gene expression and DNA copy number profiles of adipose tissue-derived stromal cells and consecutive neurosphere-like cells generated from dogs with naturally occurring spinal cord injury AU - Lim, J. H. AU - Koh, S. AU - Thomas, R. AU - Breen, M. AU - Olby, N. J. T2 - American Journal of Veterinary Research AB - Abstract OBJECTIVE To evaluate gene expression and DNA copy number in adipose tissue-derived stromal cells (ADSCs) and in ADSC-derived neurosphere-like cell clusters (ADSC-NSCs) generated from tissues of chronically paraplegic dogs. ANIMALS 14 client-owned paraplegic dogs. PROCEDURES Dorsal subcutaneous adipose tissue (< 1 cm 3 ) was collected under general anesthesia; ADSCs were isolated and cultured. Third-passage ADSCs were cultured in neural cell induction medium to generate ADSC-NSCs. Relative gene expression of mesenchymal cell surface marker CD90 and neural progenitor marker nestin was assessed in ADSCs and ADSC-NSCs from 3 dogs by quantitative real-time PCR assay; expression of these and various neural lineage genes was evaluated for the same dogs by reverse transcription PCR assay. Percentages of cells expressing CD90, nestin, glial fibrillary acidic protein (GFAP), and tubulin β 3 class III (TUJ1) proteins were determined by flow cytometry for all dogs. The DNA copy number stability (in samples from 6 dogs) and neural cell differentiation (14 dogs) were assessed with array-comparative genomic hybridization analysis and immunocytochemical evaluation, respectively. RESULTS ADSCs and ADSC-NSCs expressed neural cell progenitor and differentiation markers; GFAP and microtubule-associated protein 2 were expressed by ADSC-NSCs but not ADSCs. Relative gene expression of CD90 and nestin was subjectively higher in ADSC-NSCs than in ADSCs. Percentages of ADSC-NSCs expressing nestin, GFAP, and TUJ1 proteins were substantially higher than those of ADSCs. Cells expressing neuronal and glial markers were generated from ADSC-NSCs and had no DNA copy number instability detectable by the methods used. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested ADSCs can potentially be a safe and clinically relevant autologous source for canine neural progenitor cells. Further research is needed to verify these findings. DA - 2017/// PY - 2017/// DO - 10.2460/ajvr.78.3.371 VL - 78 IS - 3 SP - 371-380 ER - TY - JOUR TI - A Cytoplasmic RNA Virus Alters the Function of the Cell Splicing Protein SRSF2 AU - Rivera-Serrano, Efrain E. AU - Fritch, Ethan J. AU - Scholl, Elizabeth H. AU - Sherry, Barbara T2 - JOURNAL OF VIROLOGY AB - ABSTRACT To replicate efficiently, viruses must create favorable cell conditions and overcome cell antiviral responses. We previously reported that the reovirus protein μ2 from strain T1L, but not strain T3D, represses one antiviral response: alpha/beta interferon signaling. We report here that T1L, but not T3D, μ2 localizes to nuclear speckles, where it forms a complex with the mRNA splicing factor SRSF2 and alters its subnuclear localization. Reovirus replicates in cytoplasmic viral factories, and there is no evidence that reovirus genomic or messenger RNAs are spliced, suggesting that T1L μ2 might target splicing of cell RNAs. Indeed, RNA sequencing revealed that reovirus T1L, but not T3D, infection alters the splicing of transcripts for host genes involved in mRNA posttranscriptional modifications. Moreover, depletion of SRSF2 enhanced reovirus replication and cytopathic effect, suggesting that T1L μ2 modulation of splicing benefits the virus. This provides the first report of viral antagonism of the splicing factor SRSF2 and identifies the viral protein that determines strain-specific differences in cell RNA splicing. IMPORTANCE Efficient viral replication requires that the virus create favorable cell conditions. Many viruses accomplish this by repressing specific antiviral responses. We demonstrate here that some mammalian reoviruses, RNA viruses that replicate strictly in the cytoplasm, express a protein variant that localizes to nuclear speckles, where it targets a cell mRNA splicing factor. Infection with a reovirus strain that targets this splicing factor alters splicing of cell mRNAs involved in the maturation of many other cell mRNAs. Depletion of this cell splicing factor enhances reovirus replication and cytopathic effect. Our results provide the first evidence of viral antagonism of this splicing factor and suggest that downstream consequences to the cell are global and benefit the virus. DA - 2017/4// PY - 2017/4// DO - 10.1128/jvi.02488-16 VL - 91 IS - 7 SP - SN - 1098-5514 KW - RNA processing KW - SRSF KW - reovirus KW - splicing ER - TY - JOUR TI - Synthesis, photophysics and electronic structure of oxobacteriochlorins AU - Liu, Mengran AU - Chen, Chih-Yuan AU - Hood, Don AU - Taniguchi, Masahiko AU - Diers, James R. AU - Bocian, David F. AU - Holten, Dewey AU - Lindsey, Jonathan S. T2 - NEW JOURNAL OF CHEMISTRY AB - Synthetic oxobacteriochlorins exhibit strong absorption in the deep-red window flanked by chlorins to the red and bacteriochlorins to the near-infrared. DA - 2017/5/21/ PY - 2017/5/21/ DO - 10.1039/c6nj04135c VL - 41 IS - 10 SP - 3732-3744 SN - 1369-9261 ER - TY - JOUR TI - Nasogastric tube-administered alectinib achieved long-term survival in a crizotinib-refractory nonsmall cell lung cancer patient with a poor performance status AU - Kaufman, D. L. AU - Kogelnik, A. M. AU - Mozayeni, R. B. AU - Cherry, N. A. AU - Breitschwerdt, E. B. T2 - Clinical Case Reports DA - 2017/// PY - 2017/// VL - 5 IS - 6 SP - 931-935 ER - TY - JOUR TI - Multi-step excitation energy transfer engineered in genetic fusions of natural and synthetic light-harvesting proteins AU - Mancini, J. A. AU - Kodali, G. AU - Jiang, J. B. AU - Reddy, K. R. AU - Lindsey, J. S. AU - Bryant, D. A. AU - Dutton, P. L. AU - Moser, C. C. T2 - Journal of the Royal Society Interface DA - 2017/// PY - 2017/// VL - 14 IS - 127 ER - TY - JOUR TI - Malignant canine mammary tumours: Preliminary genomic insights using oligonucleotide array comparative genomic hybridisation analysis AU - Santos, Marta AU - Dias-Pereira, Patricia AU - Williams, Christina AU - Lopes, Carlos AU - Breen, Matthew T2 - VETERINARY JOURNAL AB - Neoplastic mammary disease in female dogs represents a major health concern for dog owners and veterinarians, but the genomic basis of the disease is poorly understood. In this study, we performed high resolution oligonucleotide array comparative genomic hybridisation (oaCGH) to assess genome wide DNA copy number changes in 10 malignant canine mammary tumours from seven female dogs, including multiple tumours collected at one time from each of three female dogs. In all but two tumours, genomic imbalances were detected, with losses being more common than gains. Canine chromosomes 9, 22, 26, 27, 34 and X were most frequently affected. Dissimilar oaCGH ratio profiles were observed in multiple tumours from the same dogs, providing preliminary evidence for probable independent pathogenesis. Analysis of adjacent samples of one tumour revealed regional differences in the number of genomic imbalances, suggesting heterogeneity within tumours. DA - 2017/4// PY - 2017/4// DO - 10.1016/j.tvjl.2017.03.005 VL - 222 SP - 68-71 SN - 1532-2971 KW - Array comparative genomic hybridisation KW - Canine mammary tumours KW - Chromosome KW - Genetics ER - TY - JOUR TI - Interactions between CXCR4 and CXCL12 promote cell migration and invasion of canine hemangiosarcoma AU - Im, K. S. AU - Graef, A. J. AU - Breen, M. AU - Lindblad-Toh, K. AU - Modiano, J. F. AU - Kim, J. -H. T2 - VETERINARY AND COMPARATIVE ONCOLOGY AB - The CXCR4/CXCL12 axis plays an important role in cell locomotion and metastasis in many cancers. In this study, we hypothesized that the CXCR4/CXCL12 axis promotes migration and invasion of canine hemangiosarcoma (HSA) cells. Transcriptomic analysis across 12 HSA cell lines and 58 HSA whole tumour tissues identified heterogeneous expression of CXCR4 and CXCL12, which was associated with cell movement. In vitro, CXCL12 promoted calcium mobilization, cell migration and invasion that were directly proportional to surface expression of CXCR4; furthermore, these responses proved sensitive to the CXCR4 antagonist, AMD3100, in HSA cell lines. These results indicate that CXCL12 potentiates migration and invasion of canine HSA cells through CXCR4 signalling. The direct relationship between these responses in HSA cells suggests that the CXCR4/CXCL12 axis contributes to HSA progression. DA - 2017/6// PY - 2017/6// DO - 10.1111/vco.12165 VL - 15 IS - 2 SP - 315-327 SN - 1476-5829 KW - canine hemangiosarcoma KW - CXCR4 KW - CXCL12 KW - invasion KW - migration ER - TY - JOUR TI - Hydrogen Evolution Catalysis by a Sparsely Substituted Cobalt Chlorin AU - Maher, Andrew G. AU - Passard, Guillaume AU - Dogutan, Dilek K. AU - Halbach, Robert L. AU - Anderson, Bryce L. AU - Gagliardi, Christopher J. AU - Taniguchi, Masahiko AU - Lindsey, Jonathan S. AU - Nocera, Daniel G. T2 - ACS CATALYSIS AB - A sparsely substituted chlorin macrocycle containing a Co(II) center (1-Co) has been synthesized and structurally characterized. The Co(II) atom resides in a square planar coordination environment and induces significant out-of-plane distortion of the chlorin macrocycle. The paramagnetic Co(II) center resides in the macrocycle in a S = 1/2 spin state, which displays an axial doublet signal (g⊥ ≈ 2.3 and g∥ ≈ 2.03) in the X-band EPR spectrum. The open-shell d-orbital configuration is manifest in the transient absorption spectrum, which reveals an excited-state lifetime of 8.6 ± 0.2 ps for 1-Co. The Co(II) chlorin exhibits a rich oxidation–reduction chemistry with five reversible one-electron waves (three oxidative processes and two reductive processes) observed in the cyclic voltammogram. The reduction processes of 1-Co drive hydrogen evolution catalysis. Electrochemical kinetics analysis of HER by 1-Co in trifluoroacetic acid reveals a hydrogen evolution mechanism that proceeds by an ECEC mechanism. Benchmarking the catalytic activity, 1-Co exhibits higher HER activity at low overpotentials, versus its porphyrin congeners. DA - 2017/5// PY - 2017/5// DO - 10.1021/acscatal.7b00969 VL - 7 IS - 5 SP - 3597-3606 SN - 2155-5435 KW - electrocatalysis KW - macrocycle KW - chlorin KW - cobalt KW - hydrogen evolution reaction KW - solar fuels ER - TY - JOUR TI - Effect of different headspace concentrations of bornyl acetate on fecundity ofgreen peach aphid and balsam woolly adelgid AU - Bucholz, Ethan AU - Frampton, John AU - Jetton, Robert AU - Tilotta, David AU - Lucia, Lucian T2 - SCANDINAVIAN JOURNAL OF FOREST RESEARCH AB - Balsam woolly adelgid (Adelges piceae) (Hemiptera: Adelgidae) (BWA) is an exotic pest introduced from Europe to North America in the early 1900s. Subsequent introductions and spread have enabled this pest to infest native Fraser fir stands in the Southern Appalachians and become a troublesome pest for the region’s Christmas tree industry. Means to study its fecundity and control it are consequently of high importance. Headspace solid phase micro-extraction coupled with gas chromatography and mass spectrometry were used to compare chemical differences in stem tissue between a resistant species, Veitch fir (Abies veitchii) and the susceptible Fraser fir (Abies fraseri). Comparisons demonstrated that bornyl acetate (BA), a terpenoid, was qualitatively more abundant in resistant Veitch fir than Fraser fir. Varying headspace concentrations of BA were tested to ascertain any biological impacts on egg eclosion of BWA, as well as fecundity of green peach aphid (Myzus persicae) (GPA), an insect serving as a proxy. Varying concentrations of BA and a known number of adelgid eggs did not indicate any impact of concentration on egg eclosion success. However, defoliated Veitch fir branches in treatment jars produced a significant negative impact on BWA eclosion success. Implications of these findings are discussed. DA - 2017/// PY - 2017/// DO - 10.1080/02827581.2016.1275769 VL - 32 IS - 5 SP - 397-405 SN - 1651-1891 KW - Invasive insects KW - chemical biology KW - Fraser fir KW - Veitch fir KW - Adelges piceae KW - Myzus persicae ER - TY - JOUR TI - Death of Military Working Dogs Due to Bartonella vinsonii Subspecies berkhoffii Genotype III Endocarditis and Myocarditis AU - Shelnutt, Leslie M. AU - Balakrishnan, Nandhakumar AU - DeVanna, Justin AU - Batey, Kenneth L. AU - Breitschwerdt, Edward B. T2 - MILITARY MEDICINE AB - Objectives: As a result of extensive field-related activities, military working dogs (MWDs) have an increased occupational risk for acquiring vector-borne infectious diseases. Methods: Indirect fluorescent antibody, Bartonella alpha-proteobacteria growth medium enrichment culture, and 16-23S Bartonella intergenic transcribed spacer polymerase chain reaction were performed using blood, serum, or tissue specimens. Results: Endocarditis was diagnosed in three MWDs infected with Bartonella vinsonii subspecies (subsp.) berkhoffii genotype III. One dog was also infected with Bartonella rochalimae. Conclusions: B. vinsonii subsp. berkhoffii genotype III may represent an occupational risk for MWDs that develop endocarditis or myocarditis. Comprehensive periodic screening for canine vector-borne infections, in particular occult infections caused by Bartonella spp, is prudent to avoid serious or life-threating illnesses. DA - 2017/// PY - 2017/// DO - 10.7205/milmed-d-16-00125 VL - 182 IS - 3-4 SP - E1864-E1869 SN - 1930-613X ER - TY - JOUR TI - Bartonella henselae in canine cavitary effusions: prevalence, identification, and clinical associations AU - Weeden, Amy L. AU - Cherry, Natalie A. AU - Breitschwerdt, Edward B. AU - Cheves, Avery G. AU - Wamsley, Heather L. T2 - Veterinary Clinical Pathology AB - Background Previous reports suggest an association between Bartonella infection and effusions in dogs and human beings. Objectives The aims of this study were to determine the prevalence of Bartonella infection in canine effusions and to investigate historic and clinical parameters predictive of Bartonella in dogs with effusions. Methods Canine cavitary effusions submitted for analysis and, if available, paired EDTA blood, were screened for Bartonella infection using the Bartonella α‐proteobacteria growth medium enrichment culture/ PCR diagnostic platform ( Bartonella enrichment PCR or ePCR ) at Galaxy Diagnostics, Inc. Results Bartonella henselae DNA was PCR ‐amplified and sequenced from 15% (12/80) of sampled dogs. Enrichment culture prior to PCR testing was required for Bartonella detection in 92% (11/12) of cases. Twenty percent (4/20), 13% (8/60), and 0% (0/4) of dogs with pleural, peritoneal, and pericardial effusions, respectively, tested positive. Bartonella henselae was detected most frequently in the fall, and young and middle‐aged dogs appeared to be overrepresented. Golden Retrievers and Yorkshire/Silky Terriers each comprised 25% of infected dogs (odds ratio 3.4 for Golden Retrievers). There was a weak association with hemorrhagic effusions. Fifty percent of Bartonella ‐positive dogs had hemorrhage as a component of their effusion compared to 37% of PCR ‐negative dogs (odds ratio 1.7). Conclusions Viable B henselae organisms occur in pleural and peritoneal effusions of dogs; the clinical relevance of which remains unclear and may represent opportunistic infection. Associations found in this study included seasonal variation, age, breed, and site of effusion. DA - 2017/3/6/ PY - 2017/3/6/ DO - 10.1111/vcp.12469 VL - 46 IS - 2 SP - 326-330 J2 - Vet Clin Pathol LA - en OP - SN - 0275-6382 UR - http://dx.doi.org/10.1111/vcp.12469 DB - Crossref KW - Bartonellosis KW - dog KW - peritoneal effusion KW - pleural effusion KW - seasonal variation ER - TY - JOUR TI - Synthesis and photophysical characterization of bacteriochlorins equipped with integral swallowtail substituents AU - Liu, Yizhou AU - Allu, Srinivasarao AU - Reddy, Muthyala Nagarjuna AU - Hood, Don AU - Diers, James R. AU - Bocian, David F. AU - Holten, Dewey AU - Lindsey, Jonathan S. T2 - NEW JOURNAL OF CHEMISTRY AB - The two pyrroline units of bacteriochlorins can now bear gem-dialkyl or diaryl groups (L), which project above and below the macrocycle plane, whereas dimethyl groups generally have been accessible previously. DA - 2017/6/7/ PY - 2017/6/7/ DO - 10.1039/c7nj00499k VL - 41 IS - 11 SP - 4360-4376 SN - 1369-9261 ER - TY - JOUR TI - Response of Turkish and Trojan fir to Phytophthora cinnamomi and P.cryptogea AU - Kohlway, W. H. AU - Whetten, R. W. AU - Benson, D. M. AU - Frampton, J. T2 - SCANDINAVIAN JOURNAL OF FOREST RESEARCH AB - Phytophthora root rot, primarily caused by the oomycete Phytophthora cinnamomi, is a large problem for the Christmas tree industry in North Carolina. Fraser fir (Abies fraseri) has no known innate resistance to this pathogen while some exotic fir species, such as Trojan (Abies equi-trojani) and Turkish (Abies bornmuelleriana) fir display varying amounts of resistance. A Phytophthora-resistance screening trial was completed with 1600 seedlings from 12 Turkish and Trojan fir families and Fraser and momi fir (A. firma). Seedlings from each family or species were inoculated with each of eight Phytophthora isolates, six P. cinnamomi and two Phytophthora cryptogea, in an effort to describe variability in isolate aggressiveness. Mortality was assessed as percent shoot necrosis bi-weekly for 16 weeks. Overall, rankings of resistance in fir species confirmed previous single-isolate-based results; momi fir was the most resistant, followed by Turkish, Trojan, and Fraser fir. P. cinnamomi isolates were generally more aggressive than P. cryptogea isolates. The two P. cryptogea isolates resulted in 5.6% and 0.8% mortality on Turkish fir, and 10.9% and 6.7% mortality on Trojan fir, the first reported resistance screen of these host-pathogen combinations. Pearson’s correlation testing identified a high degree of correlation between most isolates and overall mean mortality. Turkish and Trojan fir families appear to possess resistance to Phytophthora species common in North Carolina. DA - 2017/// PY - 2017/// DO - 10.1080/02827581.2017.1280076 VL - 32 IS - 5 SP - 406-411 SN - 1651-1891 KW - Abies KW - disease resistance KW - North Carolina KW - root rot ER - TY - JOUR TI - Increased diversity of Phytophthora species in Fraser fir Christmas tree plantations in the Southern Appalachians AU - Pettersson, M. AU - Frampton, J. AU - Ronnberg, J. AU - Shew, H. D. AU - Benson, D. M. AU - Kohlway, W. H. AU - Escanferla, M. E. AU - Cubeta, M. A. T2 - SCANDINAVIAN JOURNAL OF FOREST RESEARCH AB - Phytophthora root rot (PRR) disease afflicts significant economic losses to the Fraser fir Christmas tree industry. In previous surveys conducted in 1972 and from 1997 to 1998 in North Carolina, the incidence of PRR was ∼9.5% with Phytophthora cinnamomi identified as the predominant causal species isolated from infected roots of Fraser fir. Due to increased use of out-of-state planting stock since 2000, we suspected increased diversity of Phytophthora species. During 2014, we surveyed Fraser fir Christmas tree plantations in the Southern Appalachians of North Carolina, Tennessee and Virginia to determine the occurrence of pathogenic root-rotting species of Phytophthora. A weighted sampling strategy based on Christmas tree acreage was deployed to collect symptomatic Fraser fir roots from 103 commercial production fields in 14 counties. Six species of Phytophthora were isolated from infected roots sampled from 82 sites in 13 counties. Phytophthora cinnamomi, P. cryptogea and P. pini represented 70.3%, 23.1% and 1.1% of the 91 isolates. Phytophthora citrophthora, P. europaea and P. sansomeana accounted for the remaining 5.5% of the isolates and have not been identified in previously published Fraser fir surveys conducted in the region. The pathogenicity of P. citrophthora on Fraser fir was confirmed based on completion of Koch’s postulates. DA - 2017/// PY - 2017/// DO - 10.1080/02827581.2016.1265144 VL - 32 IS - 5 SP - 412-420 SN - 1651-1891 KW - Abies fraseri KW - North Carolina KW - Phytophthora cinnamomi KW - root rot KW - survey ER - TY - JOUR TI - Construction of the Bacteriochlorin Macrocycle with Concomitant Nazarov Cyclization To Form the Annulated Isocyclic Ring: Analogues of Bacteriochlorophyll a AU - Zhang, Shaofei AU - Lindsey, Jonathan S. T2 - JOURNAL OF ORGANIC CHEMISTRY AB - Bacteriochlorophylls contain a bacteriochlorin macrocycle bearing an annulated fifth ring. The fifth ring, termed the isocyclic ring or ring E, is equipped with 131-oxo and 132-carbomethoxy substituents. Herein, a general route to stable, synthetic bacteriochlorophyll analogues is described. Knoevenagel condensation (∼40 mM, rt, CH2Cl2, piperidine/AcOH/molecular sieves) of a dihydrodipyrrin–carboxaldehyde (AD half) and a dihydrodipyrrin substituted with a β-ketoester (BC half) forms a propenone bearing the two halves (a hydrobilin analogue). Subsequent treatment (0.2 mM) with acid (Yb(OTf)3, CH3CN, 80 °C) promotes a double ring-closure process: (i) condensation between the α-position of pyrrole ring A and the α-acetal unit attached to pyrroline ring B forms the bacteriochlorin macrocycle, and (ii) Nazarov cyclization of the β-(propenoyl)-substituted ring C forms the isocyclic ring (E). Five new bacteriochlorins bearing various substituents (alkyl/alkyl, aryl, and alkyl/ester) at positions 2 and 3 (β-pyrrole sites, ring A) and 132 carboalkoxy groups (R = Me or Et) were constructed in 37–61% yield from the hydrobilin analogues. The BC half and AD half are available in five and eight steps, respectively, from the corresponding pyrrole-2-carboxaldehyde and unsaturated ketone. The bacteriochlorins exhibit absorption spectra typical of bacteriopheophytins (free base bacteriochlorophylls), with a strong near-infrared absorption band (707–751 nm). DA - 2017/3/3/ PY - 2017/3/3/ DO - 10.1021/acs.joc.6b02878 VL - 82 IS - 5 SP - 2489-2504 SN - 0022-3263 ER - TY - JOUR TI - Vibrio Phage KVP40 Encodes a Functional NAD(+) Salvage Pathway AU - Lee, Jae Yun AU - Li, Zhiqun AU - Miller, Eric S. T2 - JOURNAL OF BACTERIOLOGY AB - ABSTRACT The genome of T4-type Vibrio bacteriophage KVP40 has five genes predicted to encode proteins of pyridine nucleotide metabolism, of which two, nadV and natV , would suffice for an NAD + salvage pathway. NadV is an apparent nicotinamide phosphoribosyltransferase (NAmPRTase), and NatV is an apparent bifunctional nicotinamide mononucleotide adenylyltransferase (NMNATase) and nicotinamide-adenine dinucleotide pyrophosphatase (Nudix hydrolase). Genes encoding the predicted salvage pathway were cloned and expressed in Escherichia coli , the proteins were purified, and their enzymatic properties were examined. KVP40 NadV NAmPRTase is active in vitro , and a clone complements a Salmonella mutant defective in both the bacterial de novo and salvage pathways. Similar to other NAmPRTases, the KVP40 enzyme displayed ATPase activity indicative of energy coupling in the reaction mechanism. The NatV NMNATase activity was measured in a coupled reaction system demonstrating NAD + biosynthesis from nicotinamide, phosphoribosyl pyrophosphate, and ATP. The NatV Nudix hydrolase domain was also shown to be active, with preferred substrates of ADP-ribose, NAD + , and NADH. Expression analysis using reverse transcription-quantitative PCR (qRT-PCR) and enzyme assays of infected Vibrio parahaemolyticus cells demonstrated nadV and natV transcription during the early and delayed-early periods of infection when other KVP40 genes of nucleotide precursor metabolism are expressed. The distribution and phylogeny of NadV and NatV proteins among several large double-stranded DNA (dsDNA) myophages, and also those from some very large siphophages, suggest broad relevance of pyridine nucleotide scavenging in virus-infected cells. NAD + biosynthesis presents another important metabolic resource control point by large, rapidly replicating dsDNA bacteriophages. IMPORTANCE T4-type bacteriophages enhance DNA precursor synthesis through reductive reactions that use NADH/NADPH as the electron donor and NAD + for ADP-ribosylation of proteins involved in transcribing and translating the phage genome. We show here that phage KVP40 encodes a functional pyridine nucleotide scavenging pathway that is expressed during the metabolic period of the infection cycle. The pathway is conserved in other large, dsDNA phages in which the two genes, nadV and natV , share an evolutionary history in their respective phage-host group. DA - 2017/5// PY - 2017/5// DO - 10.1128/jb.00855-16 VL - 199 IS - 9 SP - SN - 1098-5530 KW - NAD KW - bacteriophages KW - genomics KW - metabolism ER - TY - JOUR TI - Tissue and cell-type co-expression networks of transcription factors and wood component genes in Populus trichocarpa AU - Shi, Rui AU - Wang, Jack P. AU - Lin, Ying-Chung AU - Li, Quanzi AU - Sun, Ying-Hsuan AU - Chen, Hao AU - Sederoff, Ronald R. AU - Chiang, Vincent L. T2 - PLANTA DA - 2017/5// PY - 2017/5// DO - 10.1007/s00425-016-2640-1 VL - 245 IS - 5 SP - 927-938 SN - 1432-2048 KW - Wood formation KW - Transcriptome KW - Fiber cells KW - Vessel elements KW - Cell wall biosynthesis KW - Co-expression network ER - TY - JOUR TI - NF-kappa B activation is cell type-specific in the heart AU - Rivera-Serrano, Efrain E. AU - Sherry, Barbara T2 - VIROLOGY AB - Viral myocarditis is common and can progress to cardiac failure. Cardiac cell pro-inflammatory responses are critical for viral clearance, however sustained inflammatory responses contribute to cardiac damage. The transcription factor NF-κB regulates expression of many pro-inflammatory cytokines, but basal and induced activation of NF-κB in different cardiac cell types have not been compared. Here, we used primary cultures of cardiac myocytes and cardiac fibroblasts to identify cardiac cell type-specific events. We show that while viral infection readily stimulates activation of NF-κB in cardiac fibroblasts, cardiac myocytes are largely recalcitrant to activation of NF-κB. Moreover, we show that cardiac myocyte subpopulations differ in their NF-κB subcellular localization and identify the cis-Golgi as a cardiac myocyte-specific host compartment. Together, results indicate that NF-κB-dependent signaling in the heart is cardiac cell type-specific, likely reflecting mechanisms that have evolved to balance responses that can be either protective or damaging to the heart. DA - 2017/2// PY - 2017/2// DO - 10.1016/j.virol.2016.12.022 VL - 502 SP - 133-143 SN - 0042-6822 KW - NF-kappa B KW - Myocarditis KW - Cardiac myocyte KW - Cardiac fibroblast KW - Cardiomyocyte KW - Golgi KW - Reovirus KW - Heart ER - TY - JOUR TI - Genomic profiling of canine mast cell tumors identifies DNA copy number aberrations associated with KIT mutations and high histological grade AU - Mochizuki, Hiroyuki AU - Thomas, Rachael AU - Moroff, Scott AU - Breen, Matthew T2 - CHROMOSOME RESEARCH DA - 2017/6// PY - 2017/6// DO - 10.1007/s10577-016-9543-7 VL - 25 IS - 2 SP - 129-143 SN - 1573-6849 KW - Cancer KW - Comparative genomic hybridization KW - Digital PCR KW - Dog KW - Mastocytosis KW - Pug ER - TY - JOUR TI - Generation of LIF-independent induced pluripotent stem cells from canine fetal fibroblasts AU - Goncalves, N. J. N. AU - Bressan, F. F. AU - Roballo, K. C. S. AU - Meirelles, F. V. AU - Xavier, P. L. P. AU - Fukumasu, H. AU - Williams, C. AU - Breen, M. AU - Koh, S. AU - Sper, R. AU - Piedrahita, J. AU - Ambrosio, C. E. T2 - THERIOGENOLOGY AB - Takahashi and Yamanaka established the first technique in which transcription factors related to pluripotency are incorporated into the genome of somatic cells to enable reprogramming of these cells. The expression of these transcription factors enables a differentiated somatic cell to reverse its phenotype to an embryonic state, generating induced pluripotent stem cells (iPSCs). iPSCs from canine fetal fibroblasts were produced through lentiviral polycistronic human and mouse vectors (hOSKM/mOSKM), aiming to obtain pluripotent stem cells with similar features to embryonic stem cells (ESC) in this animal model. The cell lines obtained in this study were independent of LIF or any other supplemental inhibitors, resistant to enzymatic procedure (TrypLE Express Enzyme), and dependent on bFGF. Clonal lines were obtained from slightly different protocols with maximum reprogramming efficiency of 0.001%. All colonies were positive for alkaline phosphatase, embryoid body formation, and spontaneous differentiation and expressed high levels of endogenous OCT4 and SOX2. Canine iPSCs developed tumors at 120 days post-injection in vivo. Preliminary chromosomal evaluations were performed by FISH hybridization, revealing no chromosomal abnormality. To the best of our knowledge, this report is the first to describe the ability to reprogram canine somatic cells via lentiviral vectors without supplementation and with resistance to enzymatic action, thereby demonstrating the pluripotency of these cell lines. DA - 2017/4/1/ PY - 2017/4/1/ DO - 10.1016/j.theriogenology.2017.01.013 VL - 92 SP - 75-82 SN - 1879-3231 KW - iPSC KW - Canine KW - Stem cells KW - Pluripotency KW - Cellular reprogramming ER - TY - JOUR TI - Constitutive activation of alternative nuclear factor kappa B pathway in canine diffuse large B-cell lymphoma contributes to tumor cell survival and is a target of new adjuvant therapies AU - Seelig, Davis M. AU - Ito, Daisuke AU - Forster, Colleen L. AU - Yoon, Una A. AU - Breen, Matthew AU - Burns, Linda J. AU - Bachanova, Veronika AU - Lindblad-Toh, Kerstin AU - Timothy D. O'Brien, AU - Schmechel, Stephen C. AU - Rizzardi, Anthony E. AU - Modiano, Jaime F. AU - Linden, Michael A. T2 - LEUKEMIA & LYMPHOMA AB - Activation of the classical nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB) pathway is a common molecular event observed in both human and canine diffuse large B-cell lymphoma (DLBCL). Although the oncogenic potential of the alternative NFκB pathway (ANFκBP) has also been recently identified in DLBCL, its precise role in tumor pathogenesis and potential as a treatment target is understudied. We hypothesized that up-regulation of the ANFκBP plays an important role in the proliferation and survival of canine DLBCL cells, and we demonstrate that the ANFκBP is constitutively active in primary canine DLBCL samples and a cell line (CLBL1). We further demonstrate that a small interfering RNA inhibits the activation of the NFκB pathway and induces apoptosis in canine DLBCL cells. In conclusion, the ANFκBP facilitates survival of canine DLBCL cells, and thus, dogs with spontaneous DLBCL can provide a useful large animal model to study therapies targeting the ANFκBP. DA - 2017/// PY - 2017/// DO - 10.1080/10428194.2016.1260122 VL - 58 IS - 7 SP - 1702-1710 SN - 1029-2403 KW - Diffuse large B-cell lymphoma KW - alternative NF-kappa B pathway KW - canine model KW - targeting therapy KW - comparative pathology ER - TY - JOUR TI - Antimicrobial use Guidelines for Treatment of Respiratory Tract Disease in Dogs and Cats: Antimicrobial Guidelines Working Group of the International Society for Companion Animal Infectious Diseases AU - Lappin, M.R. AU - Blondeau, J. AU - Boothe, D. AU - Breitschwerdt, E.B. AU - Guardabassi, L. AU - Lloyd, D.H. AU - Papich, M.G. AU - Rankin, S.C. AU - Sykes, J.E. AU - Turnidge, J. AU - Weese, J.S. T2 - Journal of Veterinary Internal Medicine AB - Respiratory tract disease can be associated with primary or secondary bacterial infections in dogs and cats and is a common reason for use and potential misuse, improper use, and overuse of antimicrobials. There is a lack of comprehensive treatment guidelines such as those that are available for human medicine. Accordingly, the International Society for Companion Animal Infectious Diseases convened a Working Group of clinical microbiologists, pharmacologists, and internists to share experiences, examine scientific data, review clinical trials, and develop these guidelines to assist veterinarians in making antimicrobial treatment choices for use in the management of bacterial respiratory diseases in dogs and cats. DA - 2017/2/10/ PY - 2017/2/10/ DO - 10.1111/jvim.14627 VL - 31 IS - 2 SP - 279-294 J2 - J Vet Intern Med LA - en OP - SN - 0891-6640 UR - http://dx.doi.org/10.1111/jvim.14627 DB - Crossref KW - Bronchitis KW - Pneumonia KW - Pyothorax KW - Rhinitis ER - TY - JOUR TI - Variation among Loblolly Pine Seed Sources across Diverse Environments in the Southeastern United States AU - Farjat, Alfredo E. AU - Chamblee, Aaron K. AU - Isik, Fikret AU - Whetten, Ross W. AU - McKeand, Steven E. T2 - FOREST SCIENCE AB - Seven seed sources of first-generation plantation selections of loblolly pine (Pinus taeda L.) were evaluated for six traits in test sites across most of its native range east of the Mississippi River in the southeastern United States. The traits evaluated were survival, height, volume, straightness, stem forking, and incidence of fusiform rust disease (caused by Cronartium quercuum [Berk.] Miyabe ex Shirai f. sp. fusiforme). At age 8 years, survival was high, with most seed sources having survival greater than 75% at all but two test sites. South Carolina Coastal and Georgia-Florida Coastal seed sources exhibited the fastest growth and most resistance to fusiform rust, whereas the Virginia seed source exhibited the slowest growth but had the best stem form. Test sites and seed source were significantly different for all traits. Seed source × site interactions (genotype × environment [G × E]) were also significant for all traits except stem forking. Low type B genetic correlation values (rB <0.67) for height, volume, and straightness suggest the presence of G × E interactions. The South Carolina Coastal and Virginia seed sources contributed disproportionally the most to G × E interactions for growth traits, but environmental contributions to G × E interactions were distributed relatively uniformly across most test sites. The results indicate that when seed sources are moved outside of their adaptive range, important G × E interactions should be expected and the difference among seed sources originating from a wide range of climates are expected to be more pronounced in older ages. DA - 2017/2// PY - 2017/2// DO - 10.5849/forsci.15-107 VL - 63 IS - 1 SP - 39-48 SN - 1938-3738 KW - Pinus taeda KW - seed source KW - provenance test KW - genotype x environment interaction KW - multienvironmental trial analysis ER - TY - JOUR TI - Synthetic Chlorins, Possible Surrogates for Chlorophylls, Prepared by Derivatization of Porphyrins AU - Taniguchi, Masahiko AU - Lindsey, Jonathan S. T2 - CHEMICAL REVIEWS AB - Chlorophylls make Earth green, are the central constituents in the engine of photosynthesis, and not surprisingly have garnered immense attention. A chlorin, the core chromophore of a chlorophyll, is a dihydroporphyrin macrocycle that contains one pyrroline ring and three pyrrole rings. The dominant method for the synthesis of chlorins has entailed the derivatization of porphyrins. The present review covers the ostensibly simple conversion of porphyrins, regardless of synthetic or biological origin, to chlorins. The period covered encompasses the entire history since the beginnings of chlorin synthetic chemistry in the early 20th century through 2015. Representative transformations include hydrogenation, cycloaddition, annulation, and diverse “breaking and mending” approaches. Altogether, the synthesis of >1000 chlorins or chlorin-like compounds (containing >50 distinct pyrroline motifs) is described. Such diversity animates the question “what structural features are essential for a chlorin to resemble chlorophyll?” To begin to address the structure–spectrum relationship, > 250 absorption spectra are provided for representative structures. The synthesis and spectral properties of the vast collection of compounds described herein are expected to illuminate the scope to which synthetic chlorins can serve as surrogates for chlorophylls and be exploited in diverse ways. DA - 2017/1/25/ PY - 2017/1/25/ DO - 10.1021/acs.chemrev.5b00696 VL - 117 IS - 2 SP - 344-535 SN - 1520-6890 ER - TY - JOUR TI - Pyogranulomatous Pancarditis with Intramyocardial Bartonella henselae San Antonio 2 (BhSA2) in a Dog AU - Donovan, T. A. AU - Fox, P. R. AU - Balakrishnan, N. AU - Ericson, M. AU - Hooker, V. AU - Breitschwerdt, E. B. T2 - JOURNAL OF VETERINARY INTERNAL MEDICINE AB - A 6-year-old, female, spayed American Pitbull terrier was presented for progressive lethargy, weakness, and anorexia of 3-weeks duration. The dog was found as a stray in New York City (NYC) approximately 4 years earlier, assessed to be 2 years of age, and in very poor condition. Pertinent past medical history included a deep hindquarter wound and minor injuries resulting from porcupine encounters. After adoption, the dog lived in NYC, spending summers and weekends in western Massachusetts, running freely on a large property with open fields and woodlands. Before presentation, the dog was healthy and had received routine vaccinations. Ticks were found by her owners on the surface of her fur but were not embedded. Initial physical examination revealed a body weight of 19 kg, pyrexia (103.6°F), and sensitivity over the epaxial muscle region. Clinical pathology abnormalities included mild hypoalbuminemia (2.4 g/dL, range 2.7–3.9 g/dL), hyperglobulinemia (5.9 g/dL, range 2.4–4.8 g/dL), and lymphopenia (847/μL, range 1060–4950/μL). Fecal flotation test results were negative. An injection of meloxicam1 (dose not provided) was given SC, and metronidazole2 (20 mg/kg PO q12h) was prescribed for 8 days. During the following week, the dog was reexamined for increasing lethargy, weakness, and pyrexia (103.4°F). Lyme quantitative C6 ELISA (Enzyme linked immunosorbent assay) was negative (<10 U/mL, range <30 U/mL)3. The dog was seroreactive to Anaplasma phagocytophilum (1 : 400) and Ehrlichia canis (1 : 25) antigens by immunofluorescence antibody immunoassay, but was not seroreactive to Rickettsia rickettsii3. Doxycycline4 (10 mg/kg PO q24h) and meloxicam1 (6 mg/kg PO q24h) were prescribed. When reexamined 4 days later, the patient's rectal temperature was 100.2°F. Multiple, nonpainful, 1–2 cm diameter, raised cutaneous masses were observed on the left thigh and right ventral abdomen. Punch biopsies were obtained from the 2 ulcerated mass lesions and immediately fixed in formalin, after which the dog was prescribed metoclopramide5 (0.3 mg/kg PO q12h) and firocoxib6 (6 mg/kg PO q24h) and referred to the Animal Medical Center (AMC). Upon presentation to the AMC, the dog was hypothermic (95.9°) and eupneic at rest. Respiratory rate and effort increased with ambulation. Thoracic auscultation revealed focal, right-sided, fine, inspiratory crackles, and a sinus arrhythmia. Femoral pulse pressure was synchronous and hypokinetic. Neurological examination revealed lethargic mentation, decreased response to stimuli, and inconsistent conscious proprioceptive deficits. Gentle abdominal palpation elicited cranial and caudal discomfort. Venous blood gas findings were consistent with a metabolic acidosis. The dog was hypoxemic (SpO2, 84%). Oscillometric blood pressure was 109/73 mmHg; however, systolic blood pressure dropped to 70 mmHg during hospitalization. Other laboratory abnormalities included neutrophilia (13.6 K/μL, range, 2.940—12.7 K/μL), thrombocytopenia (51 K/μL, range, 143–448 K/μL), increased alkaline phosphatase (587 U/L, range, 5–160 U/L), aspartate aminotransferase (76 U/L, range, 16–55 U/L) and creatine kinase (320 U/L, range, 10–200 U/L), hypoalbuminemia (1.1 g/dL, range, 2.7–3.9 g/dL), hyperglobulinemia (5.9 g/dL, range, 2.4–4.0 g/dL), hyperbilirubinemia (2.2 mg/dL, range, 0.0–0.3 mg/dL), (unconjugated 1.2 mg/dL, range, 0.0–0.2 mg/dL, conjugated 1.0 mg/dL, range, 0.0–0.1 mg/dL), azotemia (blood urea nitrogen 168 mg/dL, range, 9–31 mg/dL; creatinine 1.9 mg/dL, range, 0.5–1.5 mg/dL), and hyponatremia (124.4 mmol/L, range 135.0–148.0 mmol/L). Thoracic radiographs revealed a diffuse, alveolar pulmonary pattern. Abbreviated abdominal and thoracic ultrasound examinations detected pleural, pericardial, and peritoneal effusions. Fluid removed via abdominocentesis on 2 attempts revealed clotting blood (PCV/TS 13/5.6) and nonclotting serosanguinous fluid (PCV 3%). Cytologic evaluation of the serosanguinous fluid identified erythrocytes, few neutrophils, and band neutrophils. Punch biopsy results revealed pyogranulomatous, necrotizing dermatitis, and panniculitis. Special stains for fungus (Gomori's methenamine silver, GMS) and acid fast bacteria (Ziehl-Neelsen, ZN) were negative. Despite supportive emergency treatment, vital signs declined. The patient's owners elected euthanasia and granted permission for necropsy, 16 days after initial medical examination. The postmortem interval was between 6 and 12 hours. Approximately 350 mL of serosanguinous, pleural effusion and 60 mL of serosanguinous, pericardial effusion intermixed with fibrin were present. The heart was mildly enlarged (1.14% of body weight). Visible from the epicardium of all 4 cardiac chambers was a miliary pattern created by myriad pinpoint, multifocal to coalescing, tan to white nodular foci (Fig 1) which extended into the myocardium and endocardium. The left atrioventricular valve leaflets were thickened at the distal aspect without chordae tendineae involvement, consistent with grade II myxomatous valvular degeneration.1 The lungs were diffusely dark red, rubbery to firm, and upon sectioning, exuded serosanguinous fluid. Sections from the right middle and right cranial lung lobes sank in formalin, suggesting cellular infiltration. The abdomen contained 30 mL of serosanguinous effusion and small amounts of clotted blood (the latter presumed to be secondary to abdominocentesis). The kidneys were bilaterally enlarged and contained myriad, multifocal to coalescing, tan to light red, irregular, nodular foci, ranging from pinpoint to 1.0 cm in diameter (Figs 2, 3). Marked hepatomegaly was present (5.5% body weight), and the liver had a diffusely enhanced reticular pattern with plaques of fibrin over the capsule and diaphragm. A single porcupine quill was encased in the proximal right lateral liver lobe with no grossly apparent tissue reaction. Lymphadenomegaly was observed including the retropharyngeal, mandibular, sternal, renal, tracheobronchial, pancreaticoduodenal, and mesenteric lymph nodes. Impression smears of the kidney and heart revealed large numbers of neutrophils and macrophages, with fewer lymphocytes and plasma cells. Bacteria were not observed. Cardiac histopathology revealed multifocal to coalescing, transmural, severe, pyogranulomatous, lymphoplasmacytic myocarditis, epicarditis, and endocarditis (pancarditis) with regional cardiomyocyte degeneration and necrosis (Fig 4). There was variable inflammation in the right and left atrioventricular valves and pulmonic and aortic valves. Rare, granulomatous, pyogranulomatous, or lymphoplasmacytic vasculitis was noted. The kidneys contained extensive, multifocal to coalescing, pyogranulomatous, tubulointerstitial nephritis with degeneration, necrosis and regeneration of tubules in areas of inflammation (Fig 5). In the lungs, neutrophilic, pyogranulomatous, and lymphoplasmacytic interstitial pneumonia was present in all lobes, with necrosis, fibrin exudation, edema, alveolar histiocytosis, arteritis, periarteritis, and mild type II pneumocyte hyperplasia. Pyogranulomatous and lymphoplasmacytic arteritis and periarteritis were prominent in lung samples (Fig 6). Chronic, passive hepatic congestion was observed. Pyogranulomatous meningitis, encephalitis, anterior uveitis, scleritis, lymphadenitis, tracheitis, peritracheitis, cystitis, capsular splenitis, and mediastinal periarteritis were also noted. In addition to mild lymphadenitis, enlarged lymph nodes exhibited changes consistent with reactive hyperplasia. The adrenal glands did not exhibit adrenalitis or atrophy. Special stains for fungus (GMS), acid fast bacteria (ZN and Fite faraco), protozoa (periodic acid–Schiff), and bacteria (Brown and Brenn, Warthin-Starry) were negative on the heart, kidney, and lungs. Warthin-Starry silver stains contained abundant artifact, which hindered interpretation. DNA was extracted from fresh-frozen canine heart, formalin-fixed, paraffin-embedded (FFPE) heart, kidney, lung, lymph node, trachea, urinary bladder, adrenal glands, brain and eye tissues with DNeasy Blood and Tissue Kit7 according to the manufacturer's instructions. Elution buffer was used as a reagent control with each set of DNA extractions. DNA concentration and purity were determined with a spectrophotometer.8 Extracted DNA was stored at −20°C. Bartonella genus and B. koehlerae species-specific PCRs were performed with primers designed to amplify the 16–23S intergenic transcribed spacer (ITS) region.2 Bartonella DNA was amplified only from fresh heart tissue, whereas FFPE kidney, lung, lymph node, trachea, urinary bladder, adrenal glands, brain, and eye were PCR negative for Bartonella spp. By sequencing, Bartonella henselae San Antonio 2 (BhSA2) DNA amplified from fresh heart tissue shared 100% (444/444 base pairs) DNA sequence similarity with BhSA2 (GenBank accession number AF369529.1). Eubacterial PCR with pan oligonucleotide primers targeting the 16S rRNA gene was performed as described previously on FFPE heart, kidney, lung, lymph node, trachea, urinary bladder, adrenal glands, brain, and eye, respectively.3 Briefly, 25 μL reaction volume consisted of 10 μM forward (fD1-5-AGAGTTTGATCCTGGCTCAG-3; Escherichia coli spanning position 339–357) and reverse (rP2-5-ACGGCTACCTTGTTACGACTT-3; E. coli spanning position 519–536) primers, 5 μL template DNA and 12.5 μL of MyTaq master mix.9 The thermocycling conditions included initial denaturation (94°C for 2 minutes followed by 35 cycles of denaturation (94°C for 30 seconds), annealing (68°C for 30 seconds) extension (72°C for 1 minute), and final extension (72°C for 1 minute).3 None of the FFPE tissues tested were positive for pathogenic bacteria. Scrolls from FFPE blocks of heart, kidney, and lung were submitted for PCR testing for canine circovirus, canine herpesvirus, canine adenovirus, Blastomyces spp., Histoplasma spp., Coccidiomyocoides spp., Cryptococcus spp., Sporothrix spp., Trichophyton spp.,10 Borrelia burgdorferi, Anaplasma spp., Rickettsia spp., Ehrlichia spp., and Bartonella.11 All of these PCR results were negative. Deparaffinized floating sections (50 μm) of the dog's heart were immunostained with a mouse anti-B. henselae antibody12 at 1 : 100 dilution and a secondary antibody donkey anti-mouse conjugated to Alexa 48813 at 1 : 400 dilution by previously published protocols.4 Thirteen 0.45-μm optical sections (total Z projection = 5.85 μm) were captured by multiphoton laser scanning microscopy (Nikon A1RMP—University of Minnesota Imaging Center) with a Nikon Plan Apo LWD25X 1.10W DICN2, Melville, NY 11747, USA objective and electronic zoom = 2). Bartonella henselae organisms (green) were visualized in the myocardium (Fig 7). These organisms were frequently amorphous and nonlinear, interpreted to be present in clumps or clusters. With multiphoton microscopy, we were able to clearly separate the epifluorescent signal contribution of endogenous lipofuscin (red channel) from the B. henselae immunoreactivity (green channel).5, 6 Heart tissue incubated with only the secondary antibody (antibody control) showed no signal in the green channel (data not shown). A negative control on normal canine heart tissue was not performed. This report describes pyogranulomatous myocarditis, epicarditis, and endocarditis (pancarditis) in a dog infected with B. henselae SA2. Bartonella henselae organisms were documented in the heart using Bartonella genus-specific 16–23S ITS elements PCR amplification from fresh heart tissue. Bartonella henselae organisms were visualized with immunoreactivity with multiphoton laser confocal microscopy, a novel technique which has not been previously described for identification of Bartonella in canine tissue. This case was notable for its wide-ranging, multi-organ pathology, particularly pyogranulomatous pancarditis, nephritis, and interstitial pneumonia with tricavitary effusion. We were unable to document coinfection with other infectious organisms. Diagnostic investigations in this case were limited by the lack of bacterial culture or virus isolation on fresh tissues at the time of necropsy. Collectively, findings were suggestive of bacteremia; however, neither Bartonella spp. nor any other infectious organisms could be identified in multiple additional organs tested by PCR, including 16S rDNA eubacterial PCR for fastidious and nonfastidious bacteria. We were unable to demonstrate the colocalization of Bartonella organisms in the inflammatory foci with traditional immunohistochemistry despite the immune reaction demonstrated with confocal microscopy. Possible explanations for these discrepancies are discussed. Bartonella henselae has important one health implications and has been associated with a wide spectrum of clinical disease syndromes in dogs and humans.7-18 Bartonella spp. are Gram-negative, alpha-proteobacteria implicated as a cause of culture-negative endocarditis, arrhythmias, myocarditis, and sudden death in animals and humans.8 This fastidious, intracellular, vasculotropic bacterium is commonly referred to as a stealth pathogen and characteristically causes persistent intravascular infections.9, 10 Bartonella spp. may cause a chronic relapsing bacteremia among reservoir hosts and resultant opportunistic infections in nonreservoir hosts.9-17 Due to Bartonella's ability to cause persistent infection within numerous cell types, demonstration of causation is often challenging and requires a high index of suspicion.17 Additional, follow-up PCR performed on FFPE tissues from multiple organs was negative. The FFPE heart samples were obtained from a different location than the fresh cardiac tissue. Pathogen DNA amplification may be affected by the size of the tissue placed in formalin, the type of tissue, and the duration of formalin fixation before embedding. DNA degradation during formalin fixation has been well documented,7, 19 especially in detecting B. henselae in splenic7 and skin tissues (N. Balakrishnan, Perdergraft JS, Kolluru S, Lappin M, Breitschwerdt EB, unpublished data). Additionally, low numbers of Bartonella within some of the inflammatory lesions or the history of antibiotic treatment may have mitigated the ability to detect bacterial pathogens in the other organs tested. In this patient, Bartonella may have survived intact within the heart due to persistent intracellular or intravascular infection and periodic dissemination to other tissues.13 Bartonella has been shown to have higher bacterial loads in the heart and lymph node as compared with other parenchymal tissues,14 which may help to explain the lack of Bartonella positivity in other tissues tested.14 The cardiac gross and histologic findings of this case are similar to 2 previously reported feline cases of pyogranulomatous myocarditis and diaphragmatic myositis, in which Bartonella were demonstrated with Warthin-Starry and Steiner silver stains, Bartonella immunohistochemistry, and PCR.2 Pancardial involvement was not observed in the feline cases.2 In this case, we were unable to document the presence of Bartonella organisms in 5-μm-thick heart sections via Warthin-Starry silver stains and Bartonella immunohistochemistry. Multiphoton confocal microscopy uses 50-μm-thick floating sections and multiple Bartonella gene targets, resulting in an increased sensitivity for detecting low copy numbers of Bartonella spp. as previously documented.20, 21 Apparent clumping or clustering of the bacteria is demonstrated by the confocal microscopy image. These results are similar to the aforementioned case series of feline myocardial Bartonella infections,2 although bacterial visualization was achieved with different techniques (silver stains and immunohistochemistry). A possible explanation for the clusters is aggregation of bacteria within a biofilm matrix; however, there is no evidence to substantiate this possibility. Unfortunately, the exact location of the bacteria (intracellular or extracellular) could not be determined. With the current case, myocarditis attributed to B. henselae has now been reported in 3 mammalian genera including cats, dogs, and humans.2, 8, 13, 15, 17, 18, 22 Myocarditis was described in 2 dogs seroreactive to Bartonella vinsonii subspecies berkhoffii antigens, with PCR amplification in 1 dog; however, Bartonella bacteria were not visualized within the myocardium. Both of these dogs had concurrent endocarditis with vegetative lesions.8 The cause for the relatively few reported cases of Bartonella myocarditis is unknown and may be attributable to a truly low incidence, difficulty in establishing a definitive diagnosis, occurrence of occult disease, or unfamiliarity with this agent as a differential diagnosis.17 Pyogranulomatous inflammation involving a single organ or multiple tissues occurs with B. henselae in humans14, 16, 18, 23, 24, cats,2, 17 and dogs.14, 15, 18, 22, 25-31 Natural infection with B. henselae or B. vinsonii subsp berkhoffii in dogs has been correlated with granulomatous hepatitis, rhinitis, sialoadenitis, mediastinitis, polyarthritis, lymphadenitis, uveitis, meningitis, encephalitis, and panniculitis,18, 25-31 the latter 5 of which were observed in this case. Coinfections might contribute to complex interactions among microbial populations, aberrant host immune response, and atypical disease manifestations as compared to single pathogen infections. The dog in the present report was seroreactive to A. phagocytophilum and E. canis antigens. Numerous bacterial, fungal, and rickettsial PCR analyses of multiple FFPE tissues were negative. In a reported series of 12 dogs with cardiac arrhythmias, endocarditis, myocarditis, syncope, conduction defects, or sudden death, 11 were seroreactive to B. vinsonii subspecies berkhoffi antigens, and 7 were coinfected with other tick-transmitted pathogens including E. canis, Babesia canis, Babesia gibsonii, or spotted fever group Rickettsiae.8 Given the breed (American Pitbull terrier) in the present report, predisposition for Babesia infection is possible,32 although no hemoparasites were identified with CBC evaluation. A recent study demonstrated B. burgdorferi antigen using immunohistochemistry in 10 Boxer puppies with pyogranulomatous myocarditis.33 In this case, both serological testing and PCR were negative for Borrelia spp., although immunohistochemistry was not performed. Differential diagnoses for cardiopulmonary vasculitis in the present case include systemic bacterial, rickettsial, fungal or viral infections, autoimmune diseases; food or drug reaction; and uremia.34 Interestingly, although vasculitis is not a recognized sequel to Bartonella infection in dogs, B. henselae has been implicated in reported cases of human vasculitis.17 The source of infection in this case was undetermined. As the dog had a history of tick exposure, tick transmission is plausible. Alternatively, the wound of unknown origin, exposure to fleas, or other arthropod vectors may have been the source of infection. Although cat fleas (Ctenocephalides felis) are considered the primary vector, B. henselae DNA has also been amplified from other arthropods, which might indicate more ecologically diverse modes of transmission.35-38 In conclusion, this report describes extensive pyogranulomatous inflammation in a dog, including severe cardiac involvement. Clusters of B. henselae were demonstrated within the myocardium by immunoreactivity with multiphoton laser scanning confocal microscopy. It must be noted that the conclusions of this study are limited in determining whether Bartonella alone or in combination with other organisms were the cause of the disseminated inflammatory processes in this patient, as additional immunohistochemical and molecular tests failed to identify Bartonella or other infectious organisms within the inflamed FFPE tissues. Future studies are required to characterize the relationship between the host and Bartonella virulence for improved understanding of patterns of disease manifestation and optimization of diagnosis and treatment. The authors acknowledge Dr. Keith Linder for performing immunohistochemistry; Ms. Brittany Thomas and Dr. Ricardo Maggi in the NCSU-CVM Vector Borne Diseases Diagnostic Laboratory for PCR testing; Dr. Christian M. Leutenegger and Dr. Marko Estrada of IDEXX Laboratories, Inc., West Sacramento, CA 95605 for performing numerous PCR analyses; Ms. Amanda Ramkissoon, Ms. Nanette Ramkissoon, Ms Iveta Simanska, and IDEXX Laboratories, Inc., Grafton, MA, 01536 for data acquisition, slide preparation, and special stains; Dr. Alexandre Rousseau for review of clinical data and Ms. Barbara Hegarty for review of the manuscript. Grant support: The author disclosed receipt of the following financial support for the research, authorship, or publication of this article: The authors received financial support from the Schiff Foundation and the Caspary Institute of the Animal Medical Center and the Vector Borne Diseases Research Foundation Account at North Carolina State University College of Veterinary Medicine to support clinical and laboratory diagnostic testing. Conflict of Interest Declaration: The authors declared no potential conflict of interest with respect to the research, authorship, or publication of this article. Off-label Antimicrobial Declaration: Authors declare no off-label use of antimicrobials. DA - 2017/// PY - 2017/// DO - 10.1111/jvim.14609 VL - 31 IS - 1 SP - 142-148 SN - 1939-1676 KW - Canine KW - Myocarditis KW - Nephritis KW - Stealth KW - Vasculitis ER - TY - JOUR TI - The Porphobilinogen Conundrum in Prebiotic Routes to Tetrapyrrole Macrocycles AU - Taniguchi, Masahiko AU - Ptaszek, Marcin AU - Chandrashaker, Vanampally AU - Lindsey, Jonathan S. T2 - ORIGINS OF LIFE AND EVOLUTION OF BIOSPHERES DA - 2017/3// PY - 2017/3// DO - 10.1007/s11084-016-9506-1 VL - 47 IS - 1 SP - 93-119 SN - 1573-0875 KW - delta-aminolevulinic acid KW - Porphyrinogen KW - Porphyrin KW - DFT KW - Biosynthesis KW - Biomimetic ER - TY - JOUR TI - Soybean cyst nematode culture collections and field populations from North Carolina and Missouri reveal high incidences of infection by viruses AU - Ruark, Casey L. AU - Koenning, Stephen R. AU - Davis, Eric L. AU - Opperman, Charles H. AU - Lommel, Steven A. AU - Mitchum, Melissa G. AU - Sit, Tim L. T2 - PLOS ONE AB - Five viruses were previously discovered infecting soybean cyst nematodes (SCN; Heterodera glycines) from greenhouse cultures maintained in Illinois. In this study, the five viruses [ScNV, ScPV, ScRV, ScTV, and SbCNV-5] were detected within SCN greenhouse and field populations from North Carolina (NC) and Missouri (MO). The prevalence and titers of viruses in SCN from 43 greenhouse cultures and 25 field populations were analyzed using qRT-PCR. Viral titers within SCN greenhouse cultures were similar throughout juvenile development, and the presence of viral anti-genomic RNAs within egg, second-stage juvenile (J2), and pooled J3 and J4 stages suggests active viral replication within the nematode. Viruses were found at similar or lower levels within field populations of SCN compared with greenhouse cultures of North Carolina populations. Five greenhouse cultures harbored all five known viruses whereas in most populations a mixture of fewer viruses was detected. In contrast, three greenhouse cultures of similar descent to one another did not possess any detectable viruses and primarily differed in location of the cultures (NC versus MO). Several of these SCN viruses were also detected in Heterodera trifolii (clover cyst) and Heterodera schachtii (beet cyst), but not the other cyst, root-knot, or reniform nematode species tested. Viruses were not detected within soybean host plant tissue. If nematode infection with viruses is truly more common than first considered, the potential influence on nematode biology, pathogenicity, ecology, and control warrants continued investigation. DA - 2017/1/31/ PY - 2017/1/31/ DO - 10.1371/journal.pone.0171514 VL - 12 IS - 1 SP - SN - 1932-6203 ER - TY - JOUR TI - Bartonellosis, One Health and all creatures great and small AU - Breitschwerdt, Edward B. T2 - Veterinary Dermatology AB - Bartonellosis is a zoonotic infectious disease of worldwide distribution, caused by an expanding number of recently discovered Bartonella spp.This review serves as an update on comparative medical aspects of this disease, including the epidemiology, pathogenesis, clinical diagnosis, treatment and challenges.Of comparative medical importance, Bartonella spp. are transmitted by several arthropod vectors, including fleas, keds, lice, sand flies, ticks and, potentially, mites and spiders. Prior to 1990, there was only one named Bartonella species (B. bacilliformis), whereas there are now over 36, of which 17 have been associated with an expanding spectrum of animal and human diseases. Recent advances in diagnostic techniques have facilitated documentation of chronic bloodstream and dermatological infections with Bartonella spp. in healthy and sick animals, in human blood donors, and in immunocompetent and immunocompromised human patients. The field of Bartonella research remains in its infancy and is rich in questions, for which patient relevant answers are badly needed. Directed Bartonella research could substantially reduce a spectrum of chronic and debilitating animal and human diseases, and thereby reduce suffering throughout the world.A One Health approach to this emerging infectious disease is clearly needed to define disease manifestations, to establish the comparative infectious disease pathogenesis of this stealth pathogen, to validate effective treatment regimens and to prevent zoonotic disease transmission. DA - 2017/1/29/ PY - 2017/1/29/ DO - 10.1111/vde.12413 VL - 28 IS - 1 SP - 96-e21 J2 - Vet Dermatol LA - en OP - SN - 0959-4493 UR - http://dx.doi.org/10.1111/vde.12413 DB - Crossref ER - TY - JOUR TI - Antibacterial efficacy and cytotoxicity of low intensity direct current activated silver-titanium implant system prototype AU - Tan, Zhuo AU - Havell, Edward A. AU - Orndorff, Paul E. AU - Shirwaiker, Rohan A. T2 - BIOMETALS DA - 2017/2// PY - 2017/2// DO - 10.1007/s10534-017-9993-1 VL - 30 IS - 1 SP - 113-125 SN - 1572-8773 KW - Low intensity direct current KW - Silver-titanium implant KW - Orthopaedic application KW - Antimicrobial efficacy KW - Cytotoxicity ER -