Cellular and ultrastructural location of angiotensinogen in rat and sheep kidney

Cellular and ultrastructural location of angiotensinogen in rat and sheep kidney. Recent evidence suggests the involvement of a local renin-angiotensin system in some renal actions of angiotensin II (Ang II). In this study the renal distribution of the precursor to angiotensin formation, angiotensinogen, was investigated in rats and sheep using immunohistochemistry, immunoelectron microscopy and non-isotopic hybridization histochemistry. Immunostaining for angiotensinogen was seen in proximal tubules (PCT) of both rat and sheep kidneys. In the rat the strongest immunostaining was found in the kidneys of neonatal (1 day old) rats. Staining declined after birth. Non-isotopic hybridization histochemistry using oligodeoxynucleotide probes labeled with biotin confirmed the presence of angiotensinogen mRNA expression in PCT of the rat renal cortex. Electron microscopic immunohistochemistry using antibodies raised against rat angiotensinogen showed weak staining in the adult of granule-like structures close to the apical membrane of PCT cells. In the neonatal rat kidney, angiotensinogen immunostaining was found throughout the PCT cells and was markedly stronger than that seen in adult rat kidney. In sheep, angiotensinogen immunostaining with an antibody raised against purified ovine angiotensinogen showed staining of PCT in fetal, newborn and adult sheep kidney. The strongest immunostaining seen was in fetal sheep kidney with a decline seen after birth. Reverse transcription polymerase chain reaction (RT-PCR) showed that angiotensinogen mRNA was expressed in the sheep kidney at all ages studied. Angiotensinogen expression was higher in fetal sheep kidneys (77 day and 141 day gestation) than in adult sheep kidney. In conclusion, angiotensinogen mRNA expression was detected in both rat and sheep kidneys. Immunostaining showed angiotensinogen protein in PCT cells of the renal cortex. Angiotensinogen staining and mRNA expression is highest during development and declines in the adult

Structure of S. aureus HPPK and discovery of a new inhibitor

The first structural and biophysical data on the folate pathway enzyme and drug target, 6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase (HPPK), from the pathogenic bacterium Staphylococcus aureus is presented. HPPK is the second essential enzyme in the folate biosynthesis pathway, responsible for catalysing pyrophosphoryl transfer from cofactor (ATP) to the substrate (6-hydroxymethyl- 7,8-dihydropterin, HMDP). In-silico screening led to the discovery of a substrate competitive inhibitor, San1, which was subsequently co-crystallised with HPPK. A 1.65 Å resolution x-ray structure showed this to bind at the pterin site sharing many of the key intermolecular interactions of the substrate. ITC and SPR measurements yielded an equilibrium binding constant, Kd, of ~13 μM for San1. An IC50 of ~12 μM was determined by means of a new convenient tri-enzyme-coupled spectrophotometric assay. ITC and SPR further showed that the San1 inhibitor has no requirement for magnesium or ATP cofactor for competitive binding to the substrate site. According to 15N heteronuclear NMR measurements, the fast motion of the pterin loop (L2) is partially dampened in the ternary complex between SaHPPK, HMDP and , -methylene adenosine 5-triphosphate (AMPCPP), but the ATP loop (L3) remains mobile on the μs timescale. In contrast, for the SaHPPK/San1/AMPCPP ternary complex, loop L2 becomes rigid on the fast timescale and loop L3 becomes more ordered which are supported by a large entropic penalty associated with San1 binding as revealed by ITC. Backbone assignments and chemical shift perturbations implicate the sulphur in San1 as a likely important loop L2/L3 stabilizing mediato

The RR Lyrae Distance Scale

We review seven methods of measuring the absolute magnitude M_V of RR Lyrae stars in light of the Hipparcos mission and other recent developments. We focus on identifying possible systematic errors and rank the methods by relative immunity to such errors. For the three most robust methods, statistical parallax, trigonometric parallax, and cluster kinematics, we find M_V (at [Fe/H] = -1.6) of 0.77 +/- 0.13, 0.71 +/- 0.15, 0.67 +/- 0.10. These methods cluster consistently around 0.71 +/- 0.07. We find that Baade-Wesselink and theoretical models both yield a broad range of possible values (0.45-0.70 and 0.45-0.65) due to systematic uncertainties in the temperature scale and input physics. Main-sequence fitting gives a much brighter M_V = 0.45 +/- 0.04 but this may be due to a difference in the metallicity scales of the cluster giants and the calibrating subdwarfs. White-dwarf cooling-sequence fitting gives 0.67 +/- 0.13 and is potentially very robust, but at present is too new to be fully tested for systematics. If the three most robust methods are combined with Walker's mean measurement for 6 LMC clusters, V_{0,LMC} = 18.98 +/- 0.03 at [Fe/H] = -1.9, then mu_{LMC} = 18.33 +/- 0.08.Comment: Invited review article to appear in: `Post-Hipparcos Cosmic Candles', A. Heck & F. Caputo (Eds), Kluwer Academic Publ., Dordrecht, in press. 21 pages including 1 table; uses Kluwer's crckapb.sty LaTeX style file, enclose

The distance to the Galactic Centre based on Population-II Cepheids and RR Lyrae stars

Context: The distance to the Galactic Centre (GC) is of importance for the distance scale in the Universe. The value derived by Eisenhauer et al. (2005) of 7.62 +- 0.32 kpc based on the orbit of one star around the central black hole is shorter than most other distance estimates based on a variety of different methods. Aim: To establish an independent distance to the GC with high accuracy. To this end Population-II Cepheids are used that have been discovered in the OGLE-II and III surveys. Method: Thirty-nine Pop-II Cepheids have been monitored on 4 nights spanning 14 days. Light curves have been fitted using the known periods from the OGLE data to determine the mean K-band magnitude. It so happens that 37 RR Lyrae stars are in the field-of-views and mean K-band magnitudes are derived for this sample as well. Results: The period-luminosity relation of Pop-II Cepheids in the K-band is determined, and the derived slope of -2.24 +- 0.14 is consistent with the value derived by Matsunaga et al. (2006). Fixing the slope to their more accurate value results in a zero point, and implies a distance modulus to the GC of 14.51 +- 0.12, with an additional systematic uncertainty of 0.07 mag. Similarly, from the RR Lyrae K-band PL-relation we derive a value of 14.48 +- 0.17 (random) +- 0.07 (syst.). The two independent determinations are averaged to find 14.50 +- 0.10 (random) +- 0.07 (syst.), or 7.94 +- 0.37 +- 0.26 kpc.Comment: A&A accepte

ROTSE All Sky Surveys for Variable Stars I: Test Fields

The ROTSE-I experiment has generated CCD photometry for the entire Northern sky in two epochs nightly since March 1998. These sky patrol data are a powerful resource for studies of astrophysical transients. As a demonstration project, we present first results of a search for periodic variable stars derived from ROTSE-I observations. Variable identification, period determination, and type classification are conducted via automatic algorithms. In a set of nine ROTSE-I sky patrol fields covering about 2000 square degrees we identify 1781 periodic variable stars with mean magnitudes between m_v=10.0 and m_v=15.5. About 90% of these objects are newly identified as variable. Examples of many familiar types are presented. All classifications for this study have been manually confirmed. The selection criteria for this analysis have been conservatively defined, and are known to be biased against some variable classes. This preliminary study includes only 5.6% of the total ROTSE-I sky coverage, suggesting that the full ROTSE-I variable catalog will include more than 32,000 periodic variable stars.Comment: Accepted for publication in AJ 4/00. LaTeX manuscript. (28 pages, 11 postscript figures and 1 gif

Distances and ages of globular clusters using Hipparcos parallaxes of local subdwarfs

We discuss the impact of Population II and Globular Cluster (GCs) stars on the derivation of the age of the Universe, and on the study of the formation and early evolution of galaxies, our own in particular. The long-standing problem of the actual distance scale to Population II stars and GCs is addressed, and a variety of different methods commonly used to derive distances to Population II stars are briefly reviewed. Emphasis is given to the discussion of distances and ages for GCs derived using Hipparcos parallaxes of local subdwarfs. Results obtained by different authors are slightly different, depending on different assumptions about metallicity scale, reddenings, and corrections for undetected binaries. These and other uncertainties present in the method are discussed. Finally, we outline progress expected in the near future.Comment: Invited review article to appear in: `Post-Hipparcos Cosmic Candles', A. Heck & F. Caputo (Eds), Kluwer Academic Publ., Dordrecht, in press. 22 pages including 3 tables and 2 postscript figures, uses Kluwer's crckapb.sty LaTeX style file, enclose

CU Comae: a new field double-mode RR Lyrae, the most metal poor discovered to date

We report the discovery of a new double-mode RR Lyrae variable (RRd) in the field of our Galaxy: CU Comae. CU Comae is the sixth such RRd identified to date and is the most metal-poor RRd ever detected. Based on BVI CCD photometry spanning eleven years of observations, we find that CU Comae has periods P0=0.5441641 +/-0.0000049d and P1=0.4057605 +/-0.0000018d. The amplitude of the primary (first-overtone) period of CU Comae is about twice the amplitude of the secondary (fundamental) period. The combination of the fundamental period of pulsation P0 and the period ratio of P1/P0=0.7457 places the variable on the metal-poor side of the Petersen diagram, in the region occupied by M68 and M15 RRd's. A mass of 0.83 solar masses is estimated for CU Comae using an updated theoretical calibration of the Petersen diagram. High resolution spectroscopy (R=30,000) covering the full pulsation cycle of CU Comae was obtained with the 2.7 m telescope of the Mc Donald Observatory, and has been used to build up the radial velocity curve of the variable. Abundance analysis done on the four spectra taken near minimum light (phase: 0.54 -- 0.71) confirms the metal poor nature of CU Comae, for which we derive [Fe/H]=-2.38 +/-0.20. This value places this new RRd at the extreme metal-poor edge of the metallicity distribution of the RR Lyrae variables in our Galaxy.Comment: 21 pages including 8 Tables, Latex, 11 Figures. Accepted for publication in The Astronomical Journal, October 2000 issu

Two populations among the metal-poor field RR Lyrae stars

We compute the spatial velocity components and the galactic orbital elements for 209 metal-poor $([Fe/H] < -1.0)$ RRLyrae (ab) variable stars in the solar neighborhood using proper motions, radial velocities, and photometric distances available in the literature. We observe abrupt changes in the stellar spatial and kinematical characteristics when the peculiar velocities relative to the local standard of rest cross the threshold value, $V_{\rm pec}\approx 280$ \mbox{km s$^{-1}$}. This provides evidence that the general population of metal-poor RRLyrae stars is not uniform, and includes two spherical subsystems occupying different volumes in the Galaxy. Based on the agreement between typical parameters of corresponding subsystems of field RRLyrae stars and of the globular clusters, studied by us earlier, we conclude that metal-poor stars and globular clusters can be subdivided into two populations, but using different criteria for stars and clusters. We suppose that field stars with fast motion and clusters with redder horizontal branches constitute the spheroidal subsystem of the accreted outer halo, with is approximately two times larger in size than the first subsystem. It has absolutely no metallicity gradients, most of its stars have eccentric orbits, many stars display retrograde motion in the Galaxy, and their ages are comparatively low, supporting the hypothesis that the objects in this subsystem have an extragalactic origin.Comment: Accepted for A&A, 8 pages, 3 figure