Gene silencing in tick cell lines using small interfering or long double-stranded RNA

Gene silencing by RNA interference (RNAi) is an important research tool in many areas of biology. To effectively harness the power of this technique in order to explore tick functional genomics and tick-microorganism interactions, optimised parameters for RNAi-mediated gene silencing in tick cells need to be established. Ten cell lines from four economically important ixodid tick genera (Amblyomma, Hyalomma, Ixodes and Rhipicephalus including the sub-species Boophilus) were used to examine key parameters including small interfering RNA (siRNA), double stranded RNA (dsRNA), transfection reagent and incubation time for silencing virus reporter and endogenous tick genes. Transfection reagents were essential for the uptake of siRNA whereas long dsRNA alone was taken up by most tick cell lines. Significant virus reporter protein knockdown was achieved using either siRNA or dsRNA in all the cell lines tested. Optimum conditions varied according to the cell line. Consistency between replicates and duration of incubation with dsRNA were addressed for two Ixodes scapularis cell lines; IDE8 supported more consistent and effective silencing of the endogenous gene subolesin than ISE6, and highly significant knockdown of the endogenous gene 2I1F6 in IDE8 cells was achieved within 48 h incubation with dsRNA. In summary, this study shows that gene silencing by RNAi in tick cell lines is generally more efficient with dsRNA than with siRNA but results vary between cell lines and optimal parameters need to be determined for each experimental system

Evidence of the Importance of Host Habitat Use in Predicting the Dilution Effect of Wild Boar for Deer Exposure to Anaplasma spp

Foci of tick-borne pathogens occur at fine spatial scales, and depend upon a complex arrangement of factors involving climate, host abundance and landscape composition. It has been proposed that the presence of hosts that support tick feeding but not pathogen multiplication may dilute the transmission of the pathogen. However, models need to consider the spatial component to adequately explain how hosts, ticks and pathogens are distributed into the landscape

Induction of humoral immune response to multiple recombinant Rhipicephalus appendiculatus antigens and their effect on tick feeding success and pathogen transmission

BACKGROUND: Rhipicephalus appendiculatus is the primary vector of Theileria parva, the etiological agent of East Coast fever (ECF), a devastating disease of cattle in sub-Saharan Africa. We hypothesized that a vaccine targeting tick proteins that are involved in attachment and feeding might affect feeding success and possibly reduce tick-borne transmission of T. parva. Here we report the evaluation of a multivalent vaccine cocktail of tick antigens for their ability to reduce R. appendiculatus feeding success and possibly reduce tick-transmission of T. parva in a natural host-tick-parasite challenge model. METHODS: Cattle were inoculated with a multivalent antigen cocktail containing recombinant tick protective antigen subolesin as well as two additional R. appendiculatus saliva antigens: the cement protein TRP64, and three different histamine binding proteins. The cocktail also contained the T. parva sporozoite antigen p67C. The effect of vaccination on the feeding success of nymphal and adult R. appendiculatus ticks was evaluated together with the effect on transmission of T. parva using a tick challenge model. RESULTS: To our knowledge, this is the first evaluation of the anti-tick effects of these antigens in the natural host-tick-parasite combination. In spite of evidence of strong immune responses to all of the antigens in the cocktail, vaccination with this combination of tick and parasite antigens did not appear to effect tick feeding success or reduce transmission of T. parva. CONCLUSION: The results of this study highlight the importance of early evaluation of anti-tick vaccine candidates in biologically relevant challenge systems using the natural tick-host-parasite combination

U Geminorum: a test case for orbital parameters determination

High-resolution spectroscopy of U Gem was obtained during quiescence. We did not find a hot spot or gas stream around the outer boundaries of the accretion disk. Instead, we detected a strong narrow emission near the location of the secondary star. We measured the radial velocity curve from the wings of the double-peaked H$\alpha$ emission line, and obtained a semi-amplitude value that is in excellent agreement with the obtained from observations in the ultraviolet spectral region by Sion et al. (1998). We present also a new method to obtain K_2, which enhances the detection of absorption or emission features arising in the late-type companion. Our results are compared with published values derived from the near-infrared NaI line doublet. From a comparison of the TiO band with those of late type M stars, we find that a best fit is obtained for a M6V star, contributing 5 percent of the total light at that spectral region. Assuming that the radial velocity semi-amplitudes reflect accurately the motion of the binary components, then from our results: K_em = 107+/-2 km/s; K_abs = 310+/-5 km/s, and using the inclination angle given by Zhang & Robinson(1987); i = 69.7+/-0.7, the system parameters become: M_WD = 1.20+/-0.05 M_sun,; M_RD = 0.42+/-0.04 M_sun; and a = 1.55+/- 0.02 R_sun. Based on the separation of the double emission peaks, we calculate an outer disk radius of R_out/a ~0.61, close to the distance of the inner Lagrangian point L_1/a~0.63. Therefore we suggest that, at the time of observations, the accretion disk was filling the Roche-Lobe of the primary, and that the matter leaving the L_1 point was colliding with the disc directly, producing the hot spot at this location.Comment: 36 pages, 14 figures, ccepted for publication in A

A New Nearby Candidate Star Cluster in Ophiuchus at d = 170 pc

The recent discoveries of nearby star clusters and associations within a few hundred pc of the Sun, as well as the order of magnitude difference in the formation rates of the embedded and open cluster populations, suggests that additional poor stellar groups are likely to be found at surprisingly close distances to the Sun. Here I describe a new nearby stellar aggregate found by virtue of the parallel proper motions, similar trigonometric parallaxes, and consistent color-magnitude distribution of its early-type members. The 120 Myr-old group lies in Ophiuchus at $d$ $\simeq$ 170 pc, with its most massive member being the 4th-magnitude post-MS B8II-III star $\mu$ Oph. The group may have escaped previous notice due to its non-negligible extinction ($A_V$ $\simeq$ 0.9 mag). If the group was born with a normal initial mass function, and the nine B- and A-type systems represent a complete system of intermediate-mass stars, then the original population was probably of order $\sim$200 systems. The age and space motion of the new cluster are very similar to those of the Pleiades, $\alpha$ Per cluster, and AB Dor Moving Group, suggesting that these aggregates may have formed in the same star-forming complex some $\sim10^8$ yr ago.Comment: 23 pages, 3 figs., to appear in Nov. 2006 A

Discovery and physical characterization as the first response to a potential asteroid collision: The case of 2023 DZ2

Near-Earth asteroids (NEAs) that may evolve into impactors deserve detailed threat assessment studies. Early physical characterization of a would-be impactor may help in optimizing impact mitigation plans. We first detected NEA 2023~DZ$_{2}$ on 27--February--2023. After that, it was found to have a Minimum Orbit Intersection Distance (MOID) with Earth of 0.00005~au as well as an unusually high initial probability of becoming a near-term (in 2026) impactor. We aim to perform a rapid but consistent dynamical and physical characterization of 2023~DZ$_{2}$ as an example of a key response to mitigate the consequences of a potential impact. We use a multi-pronged approach, drawing from various methods (observational/computational) and techniques (spectroscopy/photometry from multiple instruments), and bringing the data together to perform a rapid and robust threat assessment.} The visible reflectance spectrum of 2023~DZ$_{2}$ is consistent with that of an X-type asteroid. Light curves of this object obtained on two different nights give a rotation period $P$=6.2743$\pm$0.0005 min with an amplitude $A$=0.57$\pm$0.14~mag. We confirm that although its MOID is among the smallest known, 2023~DZ$_{2}$ will not impact Earth in the foreseeable future as a result of secular near-resonant behaviour. Our investigation shows that coordinated observation and interpretation of disparate data provides a robust approach from discovery to threat assessment when a virtual impactor is identified. We prove that critical information can be obtained within a few days after the announcement of the potential impactor.Comment: Accepted for publication in Astronomy and Astrophysics, 15 page

Mergers of close primordial binaries

We study the production of main sequence mergers of tidally-synchronized primordial short-period binaries. The principal ingredients of our calculation are the angular momentum loss rates inferred from the spindown of open cluster stars and the distribution of binary properties in young open clusters. We compare our results with the expected number of systems that experience mass transfer in post-main sequence phases of evolution and compute the uncertainties in the theoretical predictions. We estimate that main-sequence mergers can account for the observed number of single blue stragglers in M67. Applied to the blue straggler population, this implies that such mergers are responsible for about one quarter of the population of halo blue metal poor stars, and at least one third of the blue stragglers in open clusters for systems older than 1 Gyr. The observed trends as a function of age are consistent with a saturated angular momentum loss rate for rapidly rotating tidally synchronized systems. The predicted number of blue stragglers from main sequence mergers alone is comparable to the number observed in globular clusters, indicating that the net effect of dynamical interactions in dense stellar environments is to reduce rather than increase the blue straggler population. A population of subturnoff mergers of order 3-4% of the upper main sequence population is also predicted for stars older than 4 Gyr, which is roughly comparable to the small population of highly Li-depleted halo dwarfs. Other observational tests are discussed.Comment: number of pages depends on font, margins, columns etc (58 with given format), 14 figures, submitted to the Astrophysical Journa

Mass Segregation in Globular Clusters

We present the results of a new study of mass segregation in two-component star clusters, based on a large number of numerical N-body simulations using our recently developed dynamical Monte Carlo code. Specifically, we follow the dynamical evolution of clusters containing stars with individual masses m_1 as well as a tracer population of objects with individual masses m_2=\mu m_1, using N=10^5 total stars. For heavy tracers, which could represent stellar remnants such as neutron stars or black holes in a globular cluster, we characterize in a variety of ways the tendency for these objects to concentrate in or near the cluster core. In agreement with simple theoretical arguments, we find that the characteristic time for this mass segregation process varies as 1/\mu. For models with very light tracers (\mu <~ 10^-2), which could represent free-floating planets or brown dwarfs, we find the expected depletion of light objects in the cluster core, but also sometimes a significant enhancement in the halo. Using these results we estimate the optical depth to gravitational microlensing by planetary mass objects or brown dwarfs in typical globular clusters. For some initial conditions, the optical depth in the halo due to very low-mass objects could be much greater than that of luminous stars. If we apply our results to M22, using the recent null detection of Sahu, Anderson, & King (2001), we find an upper limit of ~25% at the 63% confidence level for the current mass fraction of M22 in the form of very low-mass objects.Comment: Accepted for publication in ApJ. Minor revisions reflecting the new results of Sahu et al. on M22. 13 pages in emulateapj style, including 9 figures and 3 table

Two New Candidate Planets in Eccentric Orbits

Doppler measurements of two G-type main-sequence stars, HD210277 and HD168443, reveal Keplerian variations that imply the presence of companions with masses (M sin i) of 1.28 and 5.04 M_Jup and orbital periods of 437 d and 58 d, respectively. The orbits have large eccentricities of e=0.45 and e=0.54, respectively. All 9 known extrasolar planet candidates with a=0.2-2.5 AU have orbital eccentricities greater than 0.1, higher than that of Jupiter (e=0.05). Eccentric orbits may result from gravitational perturbations imposed by other orbiting planets or stars, by passing stars in the dense star-forming cluster, or by the protoplanetary disk. Based on published studies and our near-IR adaptive optics images, HD210277 appears to be a single star. However, HD168443 exhibits a long-term velocity trend consistent with a close stellar companion, as yet undetected directly.Comment: AASTeX, 31 pages including 10 Postscript figures, to appear in the Astrophysical Journal (July 1999