Nonradioactive northern blotting for the determination of acetylcholinesterase mRNA. Comparison to the radioactive technique

ABSTRACT We have identified a second resistance-nodulation-cell division (RND)-type efflux pump, AdeIJK, in clinical isolate Acinetobacter baumannii BM4454. The adeI, adeJ, and adeK genes encode, respectively, the membrane fusion, RND, and outer membrane components of the pump. AdeJ belongs to the AcrB protein family (57% identity with AcrB from Escherichia coli). mRNA analysis by Northern blotting and reverse transcription-PCR indicated that the genes were cotranscribed. Overexpression of the cloned adeIJK operon was toxic in both E. coli and Acinetobacter. The adeIJK genes were detected in all of the 60 strains of A. baumannii tested. The two latter observations suggest that the AdeIJK complex might contribute to intrinsic but not to acquired antibiotic resistance in Acinetobacter. To characterize the substrate specificity of the pump, we have constructed derivatives of BM4454 in which adeIJK (strain BM4579), adeABC (strain BM4561), or both groups of genes (strain BM4652) were inactivated by deletion-insertion. Determination of the antibiotic susceptibility of these strains and of BM4652 and BM4579, in which the adeIJK operon was provided in trans, indicated that the AdeIJK pump contributes to resistance to β-lactams, chloramphenicol, tetracycline, erythromycin, lincosamides, fluoroquinolones, fusidic acid, novobiocin, rifampin, trimethoprim, acridine, safranin, pyronine, and sodium dodecyl sulfate. The chemical structure of these molecules suggests that amphiphilic compounds are the preferred substrates. The AdeABC and AdeIJK efflux systems contributed in a more than additive fashion to tigecycline resistance.

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Determination of mRNA half-lives inCandida albicansusing thiolutin as a transcription inhibitor

Genome ◽

10.1139/g06-046 ◽

2006 ◽

Vol 49 (8) ◽

pp. 894-899 ◽

Cited By ~ 9

Author(s):

Bessie W Kebaara ◽

Lindsey E Nielsen ◽

Kenneth W Nickerson ◽

Audrey L Atkin

Keyword(s):

Saccharomyces Cerevisiae ◽

Candida Albicans ◽

Mrna Stability ◽

18S Rrna ◽

Mrna Decay ◽

Defined Medium ◽

Northern Blotting ◽

Decay Rates ◽

Transcription Inhibitor

A method for determining mRNA half-lives in the polymorphic fungus Candida albicans is described. It employs growth in a defined medium, the inhibition of transcription with thiolutin (10–20 µg/mL), and quantitative Northern blotting. The method is effective for the A72, SC5314, and CAI-4 strains of C. albicans, and for mRNAs that have a wide variety of decay rates and steady-state abundances. The range of half-lives detected (from 4–168 min) shows that this method is effective for mRNAs with widely varying half-lives. The mRNA decay rates obtained are compared with those for orthologous mRNAs from Saccharomyces cerevisiae. This procedure should work for a broad range of C. albicans strains and can be adapted to other fungal species.Key words: comparative mRNA stability, ACT1, ADH1, EFG1, PGK1, 18S rRNA, mRNA decay.

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Galactic orbits of stars

Symposium - International Astronomical Union ◽

10.1017/s0074180900105340 ◽

1966 ◽

Vol 25 ◽

pp. 93-97

Author(s):

Richard Woolley

Keyword(s):

Globular Cluster ◽

Potential Field ◽

High Velocity ◽

Velocity Vector ◽

Variable Stars ◽

The Sun ◽

Proper Motions ◽

Rr Lyrae ◽

The Galaxy

It is now possible to determine proper motions of high-velocity objects in such a way as to obtain with some accuracy the velocity vector relevant to the Sun. If a potential field of the Galaxy is assumed, one can compute an actual orbit. A determination of the velocity of the globular clusterωCentauri has recently been completed at Greenwich, and it is found that the orbit is strongly retrograde in the Galaxy. Similar calculations may be made, though with less certainty, in the case of RR Lyrae variable stars.

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Distance to SN 1987A and the LMC

Symposium - International Astronomical Union ◽

10.1017/s0074180900118959 ◽

1999 ◽

Vol 190 ◽

pp. 549-554

Author(s):

Nino Panagia

Keyword(s):

Angular Size ◽

Large Magellanic Cloud ◽

Magellanic Cloud ◽

Light Curves ◽

Distance Modulus ◽

Absolute Size ◽

Sn 1987A

Using the new reductions of the IUE light curves by Sonneborn et al. (1997) and an extensive set of HST images of SN 1987A we have repeated and improved Panagia et al. (1991) analysis to obtain a better determination of the distance to the supernova. In this way we have derived an absolute size of the ringRabs= (6.23 ± 0.08) x 1017cm and an angular sizeR″ = 808 ± 17 mas, which give a distance to the supernovad(SN1987A) = 51.4 ± 1.2 kpc and a distance modulusm–M(SN1987A) = 18.55 ± 0.05. Allowing for a displacement of SN 1987A position relative to the LMC center, the distance to the barycenter of the Large Magellanic Cloud is also estimated to bed(LMC) = 52.0±1.3 kpc, which corresponds to a distance modulus ofm–M(LMC) = 18.58±0.05.

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International determination of the total motion of the pole

Symposium - International Astronomical Union ◽

10.1017/s0074180900177824 ◽

1961 ◽

Vol 13 ◽

pp. 29-41

Author(s):

Wm. Markowitz

Keyword(s):

Secular Motion ◽

The Future

A symposium on the future of the International Latitude Service (I. L. S.) is to be held in Helsinki in July 1960. My report for the symposium consists of two parts. Part I, denoded (Mk I) was published [1] earlier in 1960 under the title “Latitude and Longitude, and the Secular Motion of the Pole”. Part II is the present paper, denoded (Mk II).

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Time and Latitude in South Africa

International Astronomical Union Colloquium ◽

10.1017/s0252921100026816 ◽

1972 ◽

Vol 1 ◽

pp. 27-38

Author(s):

J. Hers

Keyword(s):

South Africa ◽

Cape Town ◽

Astronomical Observations ◽

Royal Observatory ◽

The Republic ◽

Astronomical Determination ◽

Limited Accuracy ◽

Better Than

In South Africa the modern outlook towards time may be said to have started in 1948. Both the two major observatories, The Royal Observatory in Cape Town and the Union Observatory (now known as the Republic Observatory) in Johannesburg had, of course, been involved in the astronomical determination of time almost from their inception, and the Johannesburg Observatory has been responsible for the official time of South Africa since 1908. However the pendulum clocks then in use could not be relied on to provide an accuracy better than about 1/10 second, which was of the same order as that of the astronomical observations. It is doubtful if much use was made of even this limited accuracy outside the two observatories, and although there may – occasionally have been a demand for more accurate time, it was certainly not voiced.

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Large-scale Motion of Solar Filaments

International Astronomical Union Colloquium ◽

10.1017/s0252921100064502 ◽

2000 ◽

Vol 179 ◽

pp. 205-208

Author(s):

Pavel Ambrož ◽

Alfred Schroll

Keyword(s):

Magnetic Flux ◽

Proper Motion ◽

Time Scale ◽

Large Scale ◽

Accuracy Of Measurements ◽

Solar Filaments ◽

General Movement ◽

Scale Motion ◽

Velocity Scatter

AbstractPrecise measurements of heliographic position of solar filaments were used for determination of the proper motion of solar filaments on the time-scale of days. The filaments have a tendency to make a shaking or waving of the external structure and to make a general movement of whole filament body, coinciding with the transport of the magnetic flux in the photosphere. The velocity scatter of individual measured points is about one order higher than the accuracy of measurements.

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Practical Realization of a Reference System for Earth Dynamics by Satellite Methods

International Astronomical Union Colloquium ◽

10.1017/s0252921100051150 ◽

1975 ◽

Vol 26 ◽

pp. 341-380 ◽

Cited By ~ 5

Author(s):

R. J. Anderle ◽

M. C. Tanenbaum

Keyword(s):

Reference Frame ◽

Polar Motion ◽

Pole Position ◽

Control Points ◽

Significant Information ◽

Crustal Motion ◽

Average Reference ◽

Future Data ◽

Existing Data

AbstractObservations of artificial earth satellites provide a means of establishing an.origin, orientation, scale and control points for a coordinate system. Neither existing data nor future data are likely to provide significant information on the .001 angle between the axis of angular momentum and axis of rotation. Existing data have provided data to about .01 accuracy on the pole position and to possibly a meter on the origin of the system and for control points. The longitude origin is essentially arbitrary. While these accuracies permit acquisition of useful data on tides and polar motion through dynamio analyses, they are inadequate for determination of crustal motion or significant improvement in polar motion. The limitations arise from gravity, drag and radiation forces on the satellites as well as from instrument errors. Improvements in laser equipment and the launch of the dense LAGEOS satellite in an orbit high enough to suppress significant gravity and drag errors will permit determination of crustal motion and more accurate, higher frequency, polar motion. However, the reference frame for the results is likely to be an average reference frame defined by the observing stations, resulting in significant corrections to be determined for effects of changes in station configuration and data losses.

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Models of Thermodynamic Properties of Prominences

International Astronomical Union Colloquium ◽

10.1017/s0252921100065799 ◽

1979 ◽

Vol 44 ◽

pp. 349-355

Author(s):

R.W. Milkey

Keyword(s):

Thermodynamic Properties ◽

Mass Transport ◽

Emission Spectrum ◽

Thermodynamic Parameters ◽

Working Group ◽

Physical Processes ◽

Theoretical Concepts ◽

Group 3 ◽

Observational Techniques

The focus of discussion in Working Group 3 was on the Thermodynamic Properties as determined spectroscopically, including the observational techniques and the theoretical modeling of physical processes responsible for the emission spectrum. Recent advances in observational techniques and theoretical concepts make this discussion particularly timely. It is wise to remember that the determination of thermodynamic parameters is not an end in itself and that these are interesting chiefly for what they can tell us about the energetics and mass transport in prominences.

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