Mutations in Ehrlichia chaffeensis Causing Polar Effects in Gene Expression and Differential Host Specificities

Citation: Cheng, C. M., Nair, A. D. S., Jaworski, D. C., & Ganta, R. R. (2015). Mutations in Ehrlichia chaffeensis Causing Polar Effects in Gene Expression and Differential Host Specificities. Plos One, 10(7), 13. doi:10.1371/journal.pone.0132657Ehrlichia chaffeensis, a tick-borne rickettsial, is responsible for human monocytic ehrlichiosis. In this study, we assessed E. chaffeensis insertion mutations impacting the transcription of genes near the insertion sites. We presented evidence that the mutations within the E. chaffeensis genome at four genomic locations cause polar effects in altering gene expressions. We also reported mutations causing attenuated growth in deer (the pathogen's reservoir host) and in dog (an incidental host), but not in its tick vector, Amblyomma americanum. This is the first study documenting insertion mutations in E. chaffeensis that cause polar effects in altering gene expression from the genes located upstream and downstream to insertion sites and the differential requirements of functionally active genes of the pathogen for its persistence in vertebrate and tick hosts. This study is important in furthering our knowledge on E. chaffeensis pathogenesis

Vaccination with an Attenuated Mutant of Ehrlichia chaffeensis Induces Pathogen-Specific CD4(+) T Cell Immunity and Protection from Tick-Transmitted Wild-Type Challenge in the Canine Host

Citation: McGill, J. L., Nair, A. D. S., Cheng, C. M., Rusk, R. A., Jaworski, D. C., & Ganta, R. R. (2016). Vaccination with an Attenuated Mutant of Ehrlichia chaffeensis Induces Pathogen-Specific CD4(+) T Cell Immunity and Protection from Tick-Transmitted Wild-Type Challenge in the Canine Host. Plos One, 11(2), 15. doi:10.1371/journal.pone.0148229Ehrlichia chaffeensis is a tick-borne rickettsial pathogen and the causative agent of human monocytic ehrlichiosis. Transmitted by the Amblyomma americanum tick, E. chaffeensis also causes disease in several other vertebrate species including white-tailed deer and dogs. We have recently described the generation of an attenuated mutant strain of E. chaffeensis, with a mutation in the Ech_0660 gene, which is able to confer protection from secondary, intravenous-administered, wild-type E. chaffeensis infection in dogs. Here, we extend our previous results, demonstrating that vaccination with the Ech_0660 mutant protects dogs from physiologic, tick-transmitted, secondary challenge with wild-type E. chaffeensis; and describing, for the first time, the cellular and humoral immune responses induced by Ech_0660 mutant vaccination and wild-type E. chaffeensis infection in the canine host. Both vaccination and infection induced a rise in E. chaffeensis-specific antibody titers and a significant Th1 response in peripheral blood as measured by E. chaffeensis antigen-dependent CD4(+) T cell proliferation and IFN. production. Further, we describe for the first time significant IL-17 production by peripheral blood leukocytes from both Ech_0660 mutant vaccinated animals and control animals infected with wild-type E. chaffeensis, suggesting a previously unrecognized role for IL-17 and Th17 cells in the immune response to rickettsial pathogens. Our results are a critical first step towards defining the role of the immune system in vaccine-induced protection from E. chaffeensis infection in an incidental host; and confirm the potential of the attenuated mutant clone, Ech_0660, to be used as a vaccine candidate for protection against tick-transmitted E. chaffeensis infection

Comparative Experimental Infection Study in Dogs with Ehrlichia canis, E. chaffeensis, Anaplasma platys and A. phagocytophilum

Citation: Nair, A. D. S., Cheng, C., Ganta, C. K., Sanderson, M. W., Alleman, A. R., Munderloh, U. G., & Ganta, R. R. (2016). Comparative Experimental Infection Study in Dogs with Ehrlichia canis, E. chaffeensis, Anaplasma platys and A. phagocytophilum. Plos One, 11(2), 21. doi:10.1371/journal.pone.0148239Dogs acquire infections with the Anaplasmataceae family pathogens, E. canis, E. chaffeensis, E. ewingii, A. platys and A. phagocytophilum mostly during summer months when ticks are actively feeding on animals. These pathogens are also identified as causing diseases in people. Despite the long history of tick-borne diseases in dogs, much remains to be defined pertaining to the clinical and pathological outcomes of infections with these pathogens. In the current study, we performed experimental infections in dogs with E. canis, E. chaffeensis, A. platys and A. phagocytophilum. Animals were monitored for 42 days to evaluate infection-specific clinical, hematological and pathological differences. All four pathogens caused systemic persistent infections detectible throughout the 6 weeks of infection assessment. Fever was frequently detected in animals infected with E. canis, E. chaffeensis, and A. platys, but not in dogs infected with A. phagocytophilum. Hematological differences were evident in all four infected groups, although significant overlap existed between the groups. A marked reduction in packed cell volume that correlated with reduced erythrocytes and hemoglobin was observed only in E. canis infected animals. A decline in platelet numbers was common with E. canis, A. platys and A. phagocytophilum infections. Histopathological lesions in lung, liver and spleen were observed in all four groups of infected dogs; infection with E. canis had the highest pathological scores, followed by E. chaffeensis, then A. platys and A. phagocytophilum. All four pathogens induced IgG responses starting on day 7 post infection, which was predominantly comprised of IgG2 subclass antibodies. This is the first detailed investigation comparing the infection progression and host responses in dogs after inoculation with four pathogens belonging to the Anaplasmataceae family. The study revealed a significant overlap in clinical, hematological and pathological changes resulting from the infections

Cosmokinetics: A joint analysis of Standard Candles, Rulers and Cosmic Clocks

We study the accelerated expansion of the universe by using the kinematic approach. In this context, we parameterize the deceleration parameter, q(z), in a model independent way. Assuming three simple parameterizations we reconstruct q(z). We do the joint analysis with combination of latest cosmological data consisting of standard candles (Supernovae Union2 sample), standard ruler (CMB/BAO), cosmic clocks (age of passively evolving galaxies) and Hubble (H(z)) data. Our results support the accelerated expansion of the universe.Comment: PDFLatex, 15 pages, 12 pdf figures, revised version to appear in JCA

Absence seizures as resetting mechanisms of brain dynamics

To understand the increase in age-related incidence and frequency of absence seizures in the rat brain, we investigated the effect of these seizures on brain dynamics. This paper puts forward the hypothesis that age-related differences in the expression of absence seizures are associated with the ability of the seizures to reset brain dynamics

An efficient design of embedded memories and their testability analysis using Markov chains

This article presents a design strategy for efficient and comprehensive random testing of embedded random-access memory (RAM) where neither are the address, read/write and data input lines directly controllable nor are the data output lines externally observable. Unlike the conventional approaches, which frequently employ on-chip circuits such as linear feedback shift register (LFSR), data registers and multibit comparator for verifying the response of the memory-under-test (MUT) with the reference signature of a fault-free gold unit , the proposed technique uses an efficient testable design, which helps accelerate test algorithms by a factor of 0.5√ n , if the RAM is organized into an n ×1 array and improve the test reliability by eliminating the LFSR that is known to have aliasing problems. Another serious problem in embedded memory testing by random test patterns is the problem of memory initialization, which has been tackled here by adding word-line flag registers. The paper has made indepth empirical studies of the functional faults such as stuck-at, coupling, and pattern-sensitive by suitably representing these faults by Markov chains and by simulating these chains to derive various test lengths required for detecting these faults. The simulation results conclusively show that, in order to test a IM-bit RAM for detecting the common functional faults, the proposed technique needs only one second as opposed to about an hour needed by the conventional random testing where memory cells are tested sequentially.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/43013/1/10836_2004_Article_BF00134733.pd

Interception of the Bycroft-Gowland Intermediate in the Enzymatic Macrocyclization of Thiopeptides

Thiopeptides are a broad class of macrocyclic, heavily modified peptide natural products that are unified by the presence of a substituted, nitrogen-containing heterocycle core. Early work indicated that this core might be fashioned from two dehydroalanines by an enzyme-catalyzed aza-[4 + 2] cycloaddition to give a cyclic-hemiaminal intermediate. This common intermediate could then follow a reductive path toward a dehydropiperidine, as in the thiopeptide thiostrepton, or an aromatization path to yield the pyridine groups observed in many other thiopeptides. Although several of the enzymes proposed to perform this cycloaddition have been reconstituted, only pyridine products have been isolated and any hemiaminal intermediates have yet to be observed. Here, we identify the conditions and substrates that decouple the cycloaddition from subsequent steps and allow interception and characterization of this long hypothesized intermediate. Transition state modeling indicates that the key amide-iminol tautomerization is the major hurdle in an otherwise energetically favorable cycloaddition. An anionic model suggests that deprotonation and polarization of this amide bond by TbtD removes this barrier and provides a sufficient driving force for facile (stepwise) cycloaddition. This work provides evidence for a mechanistic link between disparate cyclases in thiopeptide biosynthesis

Role of the νg9/2 orbital in the development of collectivity in the A≈60 region: The case of Co 61

An extensive study of the level structure of Co61 has been performed following the complex Mg26(Ca48,2α4npγ)Co61 reaction at beam energies of 275, 290, and 320 MeV using Gammasphere and the Fragment Mass Analyzer (FMA). The low-spin structure is discussed within the framework of shell-model calculations using the GXPF1A effective interaction. Two quasirotational bands consisting of stretched-E2 transitions have been established up to spins I=41/2 and (43/2), and excitation energies of ∼17 and ∼20 MeV, respectively. These are interpreted as signature partners built on a neutron ν(g9/2)2 configuration coupled to a proton πp3/2 state, based on cranked shell model (CSM) calculations and comparisons with observations in neighboring nuclei. In addition, four ΔI=1 bands were populated to high spin, with the yrast dipole band interpreted as a possible candidate for the shears mechanism, a process seldom observed thus far in this mass region