Characterization of iodothyronine sulfotransferase activity in rat liver

Sulfation is an important pathway in the metabolism of thyroid hormone because it strongly facilitates the degradation of the hormone by the type I iodothyronine deiodinase. However, little is known about the properties and possible regulation of the sulfotransferase(s) involved in the sulfation of thyroid hormone. We have developed a convenient method for the analysis of iodothyronine sulfotransferase activity in tissue cytosolic fractions, using radioiodinated 3,3'-diiodothyronine (3,3'-T2) as the preferred substrate, unlabeled 3'-phosphoadenosine-5'-phosphosulfate (PAPS) as the sulfate donor, and Sephadex LH-20 minicolomns for separation of the products. We found that iodothyronine sulfotransferase activity in rat liver cytosol is 1) higher in male than in female rats; 2) optimal at pH 8.0; 3) characterized (at 50 microM PAPS and pH 7.2) by apparent Michaelis-Menton (Km) values for 3,3'-T2 of 1.77 and 4.19 microM, and Vmax values of 1.94 and 1.45 nmol/min per mg protein in male and female rats, respectively; 4) characterized (at 1 microM 3,3'-T2 and pH 7.2) by apparent Km values for PAPS of 4.92 and 3.80 microM and Vmax values of 0.72 and 0.31 nmol/min per mg protein, in males and females, respectively; 5) little affected by hyperthyroidism in both male and female rats, but significantly decreased by hypothyroidism in males but not in females; and 6) not affected by short-term (3 days) fasting in both male and female rats, but significantly decreased by long-term (3 weeks) food restriction to one-third of normal intake in males but not in females. It is suggested that the higher hepatic iodothyronine sulfotransferase activity in male vs. female rats, as well as the decreases induced in males by hypothyroidism and long-term food restriction, represents differences in the expression of the male-dominant isoenzyme rSULT1C1

Chandra observations of the bursting X-ray transient SAX J1747.0-2853 during low-level accretion activity

We present Chandra/ACIS observations of the bursting X-ray transient SAX J1747.0-2853 performed on 18 July 2001. We detected a bright source at the position of R.A = 17^h 47^m 02.60^s and Dec. = -28 52' 58.9'' (J2000.0; with a 1 sigma error of ~0.7 arcseconds), consistent with the BeppoSAX and ASCA positions of SAX J1747.0-2853 and with the Ariel V position of the transient GX +0.2,-0.2, which was active during the 1970's. The 0.5-10 keV luminosity of the source during our observations was ~3 x 10^{35} erg/s (assuming a distance of 9 kpc) demonstrating that the source was in a low-level accretion state. We also report on the long-term light curve of the source as observed with the all sky monitor aboard the Rossi X-ray Timing Explorer. After the initial 1998 outburst, two more outbursts (in 2000 and 2001) were detected with peak luminosities about two orders of magnitude larger than our Chandra luminosity. Our Chandra observation falls in-between those two outbursts, making the outburst history for SAX J1747.0-2853 complex. Those bright 2000 and 2001 outbursts combined with the likely extended period of low level activity in-between those outbursts strongly suggest that the classification of SAX J1747.0-2853 as a faint X-ray transient was premature. It might be possible that the other faint X-ray transients also can exhibit bright, extended outbursts which would eliminate the need for a separate sub-class of X-ray transients. We discuss our results also in the context of the behavior of X-ray binaries accreting at low levels with luminosities around 10^{35} erg/s, a poorly studied accretion rate regime.Comment: Accepte for publication in ApJ, 11 July 200

BeppoSAX measurements of the bright gamma-ray burst 010222

We analyze the BeppoSAX measurements of the prompt and afterglow emission of the gamma-ray burst GRB010222. Among 45 GRBs detected with the Wide Field Cameras on BeppoSAX, the 40-700 keV fluence of (9.3+/-0.3)E-5 erg cm-2 is only surpassed by GRB990123. In terms of the isotropic 20-2000 keV energy output of 7.8E53 erg, it ranks third of all GRBs with measured distances. Since this burst is so bright, the data provide complete and valuable coverage up to 65 hr after the event, except for a gap between 3.5 and 8.0 hr. The 2-10 keV flux history shows clear signs of a break which is consistent with a break seen in the optical, and provides supporting evidence for the achromatic nature of the break. An explanation for the break in the context of a collimated expansion is not straightforward. Rather, a model is favored whereby the fireball is braked to the non-relativistic regime quickly (within a fraction of day) by a dense 1E6 cm-3 circumburst medium. This implies that, after a mild beaming correction, GRB010222 may be the most energetic burst observed thus far. The X-ray decay index after the break is 1.33+/-0.04, the spectral index 0.97+/-0.05. The decay is, with unprecedented accuracy, identical to that observed in the optical.Comment: Accepted on June 6 for publication in ApJ part I. Publication due in October 2001. Accepted version has only minor modification

Influence of spin 1/2 hetero-nuclei on spin relaxation and polarization transfer among strongly coupled protons

Effects of spin-spin interactions on the nuclear magnetic relaxation dispersion (NMRD) of protons were studied in a situation where spin ½ hetero- nuclei are present in the molecule. As in earlier works [K. L. Ivanov, A. V. Yurkovskaya, and H.-M. Vieth, J. Chem. Phys.129, 234513 (2008)10.1063/1.3040272;S. E. Korchak, K. L. Ivanov, A. V. Yurkovskaya, and H.-M. Vieth, J. Chem. Phys.133, 194502 (2010)10.1063/1.3495988], spin-spin interactions have a pronounced effect on the relaxivity tending to equalize the longitudinal relaxation times once the spins become strongly coupled at a sufficiently low magnetic field. In addition, we have found influence of 19F nuclei on the proton NMRD, although in the whole field range, studied protons and fluorine spins were only weakly coupled. In particular, pronounced features in the proton NMRD were found; but each feature was predominantly observed only for particular spin states of the hetero-nuclei. The features are explained theoretically; it is shown that hetero-nuclei can affect the proton NMRD even in the limit of weak coupling when (i) protons are coupled strongly and (ii) have spin-spin interactions of different strengths with the hetero-nuclei. We also show that by choosing the proper magnetic field strength, one can selectively transfer proton spin magnetization between spectral components of choice

Thyroid function and deiodinase activities in rats with marginal iodine deficiency

The hypothesis tested was whether marginal iodine deficiency for a period of 6 wk affects iodothyronine deiodinase activities in liver and brain of rats. Male rats were fed purified diets either deficient or sufficient in iodine; the diets were fed on a restricted basis (60% of ad libitum intake). Body weight gain of the two groups was comparable. Iodine deficiency was evidenced by increased thyroid weight (26%), reduced urinary iodine excretion (80%), and reduced plasma T4 concentrations (22%). Activities of liver type I and brain type III deiodinase were unchanged, but the activity of type II deiodinase in brain was increased (28%) in the iodine-deficient rats. Food restriction per se significantly lowered T3 (30%) and T4 (22%) concentrations in plasma and decreased type III deiodinase activity in brain (30%). These results indicate that in marginal iodine deficiency the activities of hepatic type I deiodinase and brain type III deiodinase are unchanged, whereas that of brain type II deiodinase is increased

Expression of chicken hepatic type I and type III iodothyronine deiodinases during embryonic development

In embryonic chicken liver (ECL) two types of iodothyronine deiodinases are expressed: D1 and D3. D1 catalyzes the activation as well as the inactivation of thyroid hormone by outer and inner ring deiodination, respectively. D3 only catalyzes inner ring deiodination. D1 and D3 have been cloned from mammals and amphibians and shown to contain a selenocysteine (Sec) residue. We characterized chicken D1 and D3 complementary DNAs (cDNAs) and studied the expression of hepatic D1 and D3 messenger RNAs (mRNAs) during embryonic development. Oligonucleotides based on two amino acid sequences strongly conserved in the different deiodinases (NFGSCTSecP and YIEEAH) were used for reverse transcription-PCR of poly(A+) RNA isolated from embryonic day 17 (E17) chicken liver, resulting in the amplification of two 117-bp DNA fragments. Screening of an E17 chicken liver cDNA library with these probes led to the isolation of two cDNA clones, ECL1711 and ECL1715. The ECL1711 clone was 1360 bp long and lacked a translation start site. Sequence alignment showed that it shared highest sequence identity with D1s from other vertebrates and that the coding sequence probably lacked the first five nucleotides. An ATG start codon was engineered by site-directed mutagenesis, generating a mutant (ECL1711M) with four additional codons (coding for MGTR). The open reading frame of ECL1711M coded for a 249-amino acid protein showing 58-62% identity with mammalian D1s. An in-frame TGA codon was located at position 127, which is translated as Sec in the presence ofa Sec insertion sequence (SECIS) identified in the 3'-untranslated region. Enzyme activity expressed in COS-1 cells by transfection with ECL1711M showed the same catalytic, substrate, and inhibitor specificities as native chicken D1. The ECL1715 clone was 1366 bp long and also lacked a translation start site. Sequence alignment showed that it was most homologous with D3 from other species and that the coding sequence lacked approximately the first 46 nucleotides. The deduced amino acid sequence showed 62-72% identity with the D3 sequences from other species, including a putative Sec residue at a corresponding position. The 3'-untranslated region of ECL1715 also contained a SECIS element. These results indicate that ECL1711 and ECL1715 are near-full-length cDNA clones for chicken D1 and D3 selenoproteins, respectively. The ontogeny of D1 and D3 expression in chicken liver was studied between E14 and 1 day after hatching (C1). D1 activity showed a gradual increase from E14 until C1, whereas D1 mRNA level remained relatively constant. D3 activity and mRNA level were highly significantly correlated, showing an increase from E14 to E17 and a strong decrease thereafter. These results suggest that the regulation of chicken hepatic D3 expression during embryonic development occurs predominantly at the pretranslational level