Metabolism of histones in malignant tissues and liver of the rat and mouse

1. The relative amounts of incorporation in vivo of l-lysine, and in one experiment l-arginine, into different histone fractions from Krebs ascites and a lymphoma ascites cells of mice and a `solid' tumour and liver of rats have been determined. 2. No marked differences in the incorporations of the amino acids into the fractions F1, F2a, F2b and F3 from the tumours were generally observed, although in some experiments there was a greater incorporation into fraction F2b, which could be decreased by further purification. 3. In the tumours the incorporations into all cell protein fractions obtained were approximately the same, indicating that the amount of incorporation was that required for the increase of cell mass. 4. In rat liver, the incorporations into fractions F1, F2a and F3 were not greatly different. That into fraction F2b was variable. The incorporation into the histone fractions was much less than that into the acid-insoluble nuclear residue, indicating that considerable turnover of amino acids in the latter occurs. 5. The decrease in radioactivity of labelled histone and acid-insoluble nuclear protein in vivo during several days confirmed the relatively small turnover of the histone fraction. The time taken for liver whole histone to lose half its radioactivity was about 1 week. A histone fraction of slower metabolism was also detected. 6. It is concluded that no appreciable turnover of protein occurs in any one histone fraction, the somewhat higher values obtained in certain cases being associated with acidic impurities. The apparently high rate of incorporation into histone of resting liver is discussed in relation to recent evidence on DNA metabolism of resting liver.     

Characteristics of ouabain binding to isolated trout hepatocytes

Summary Na⁺, K⁺ exchanges were studied in isolated hepatocytes of the rainbow trout, Salmo gairdneri. Ouabain at 10^–4 M produced maximal inhibition (95%) of K⁺ uptake and enhanced intracellular Na⁺ accumulation, showing that active fluxes account for a very large proportion of Na⁺ and K⁺ exchanges. Inhibition of the Na–K pump by ouabain was significant at low concentrations (10^–8 M). When external K⁺ concentration was reduced from 7 mM to 0.5 mM, half maximum inhibition (IC₅₀) of K⁺ uptake was obtained at a 22-fold lower concentration of ouabain confirming that ouabain and potassium compete at the same pump site. Time-course analysis of [³H]ouabain binding indicated a two-component kinetics: one component saturable and dependent on K⁺ concentration in the medium, the other linear and independent of external K⁺. The ouabain binding site number, determined by Scatchard plots, remained constant (ca. 2.5·10⁵ per cell) and independent of the external K⁺ concentration (7, 0.5 or 0 mM), while the dissociation constant (K_D) decreased from 4.2 M to 7.3 nM when K⁺ was removed from the Hank's medium. These ouabain binding sites are characterized by an exceptionally low turnover rate (400 min^–1), as estimated from ouabain-sensitive K⁺ flux, in comparison to those described in other cell types of higher vertebrates. At each external K⁺ concentration studied, the inhibition of K⁺ uptake and ouabain binding measured as a function of ouabain concentration indicated a strict correlation between the degree of K pump inhibition and the amount of bound glycoside.     

Protection against myxomatosis and rabbit viral hemorrhagic disease with recombinant myxoma viruses expressing rabbit hemorrhagic disease virus capsid protein.

Two myxoma virus-rabbit hemorrhagic disease virus (RHDV) recombinant viruses were constructed with the SG33 strain of myxoma virus to protect rabbits against myxomatosis and rabbit viral hemorrhagic disease. These recombinant viruses expressed the RHDV capsid protein (VP60). The recombinant protein, which is 60 kDa in size, was antigenic, as revealed by its reaction in immunoprecipitation with antibodies raised against RHDV. Both recombinant viruses induced high levels of RHDV- and myxoma virus-specific antibodies in rabbits after immunization. Inoculations by the intradermal route protected animals against virulent RHDV and myxoma virus challenges.     

Certain withanolides from Iochroma gesnerioides antagonize ecdysteroid action in a Drosophila melanogaster cell line

Sixteen withanolides isolated from Iochroma gesnerioides (Kunth) Miers (Solanaceae) have been assessed for their activities as ecdysteroid agonists and antagonists. None of the compounds showed any agonistic activity, but several showed significant antagonistic activity. With a 20-hydroxyecdysone concentration of 5 × 10^–8 M, the ED₅₀ values for 2,3-dihydro-3-methoxywithaferin A, 2,3-dihydro-3-methoxywithacnistine, 2,3-dihydro-3-methoxyiochromolide and 2,3-dihydro-3-hydroxywithacnistine are 3.5 × 10^-5 M, 1 × 10^–5 M, 5 × 10^–6 M and 2.5 × 10^–6 M, respectively.     

Synthesis and characterization of SrCu2(O2CR)3(bdmap)3 and Bi2(O2CCH3)4(bdmap)2(H2O) (R = CH3, CF3; bdmap = 1,3-bis(dimethylamino)-2-propanolato)

New mixed metal complexes SrCu₂(O₂CR)₃(bdmap)₃ (R = CF₃ (1a), CH₃ (1b)) and a new dinuclear bismuth complex Bi₂(O₂CCH₃)₄(bdmap)₂(H₂O) (2) have been synthesized. Their crystal structures have been determined by single-crystal X-ray diffraction analyses. Thermal decomposition behaviors of these complexes have been examined by TGA and X-ray powder diffraction analyses. While compound 1a decomposes to SrF₂ and CuO at about 380°C, compound 1b decomposes to the corresponding oxides above 800°C. Compound 2 decomposes cleanly to Bi₂O₃ at 330°C. The magnetism of 1a was examined by the measurement of susceptibility from 5–300 K. Theoretical fitting for the susceptibility data revealed that 1a is an antiferromagnetically coupled system with g = 2.012(7), −2J = 34.0(8) cm⁻¹. Crystal data for 1a: C₂₇H₅₁N₆O₉F₉Cu₂Sr/THF, monoclinic space group P2₁/m, A = 10.708(6), B = 15.20(1), C = 15.404(7) Å, β = 107.94(4)°, V = 2386(2) ų, Z = 2; for 1b: C₂₇H₆₀N₆O₉Cu₂Sr/THF, orthorhombic space group Pbcn, A = 19.164(9), B = 26.829(8), C = 17.240(9) Å, V = 8864(5) ų, Z = 8; for 2: C₂₂H₄₈O₁₁N₄Bi₂, monoclinic space group P2₁/c, A = 17.614(9), B = 10.741(3), C = 18.910(7) Å, β = 109.99(3)°, V = 3362(2) ų, Z = 4.     

The Quaternary eolian sequence of Madeira: stratigraphy, chronology, and paleoenvironmental interpretation

A thick (ca. 40 m) sequence of coastal eolian sediments occurs on a narrow peninsula on the eastern end of the island of Madeira, located in the Eastern Atlantic at 33°N latitude. The sediments consist of black volcanic sands (with or without bioclasts) as well as clay units up to 2 m thick. A series of inceptisols (Eutrochrepts) and one alfisol (a Hapludalf) are developed in these sediments. Land snail shells and secondary carbonates, in the form of well-developed rhizoliths, calcretes, fissure-fills, and soil nodules, are present in abundance. The chronology of the sequence was determined by ¹⁴C and U---Th analyses of land snail shells and secondary carbonates and amino acid epimerization analysis of land snail shells. All sediments, including the clay units, are originally of eolian origin, derived from the beach to the south of the deposit, but some have been redeposited by colluviation. Temporal variation in the lithology of the sediments relates to variations in sea-level, with black sands being deposited during lower sea level stands and clays at the lowest. It is suggested that fine marine sediments, exposed during low sea-level stands, may also be the dominant source of silty or clayey units in other coastal eolian deposits in the subtropical Atlantic and Mediterranean.

The sequence spans from 200,000–300,000 years ago up to the 20th century. Sedimentation was discontinuous and often rapid; erosional hiatuses are present. During the Holocene, eolian sands started accumulating at 8200 yr B.P. during a transgressive phase and stopped at 4500 yr B.P. as sea level approached its present height. Colluviation increased dramatically following the first human settlement of the island in the 15th century and continued up to the 20th century, as dated by amino acid epimerization analysis of land snails. Earlier periods of colluviation were identified from the age distribution of land snail shells redeposited in younger colluvium.

Paleoenvironmental reconstruction was based mainly on soil and sediment features (including rhizolith morphology) and land snail faunas but also on stable isotope variations (¹³C, ¹⁸O) in land snails and secondary carbonates, pollen (generally not well preserved), and phytoliths. Most of the portion of the Middle Pleistocene represented in the sequence was characterized by moderately dry conditions, in comparison to the late Pleistocene and Holocene. During the last interglacial, relatively wet conditions occurred, wetter than during the Holocene interglacial. Moderately moist conditions were present during the accumulation of the thick unit dating to ca. 80,000 yr B.P. As sea level fell subsequent to this period, conditions appear to have become drier. Starting ca. 50,000–55,000 yr B.P., conditions were especially wet, but prior to the last glacial maximum, markedly arid conditions ensued. Toward the end of the last glacial, wet conditions returned and produced the best-developed soil preserved in the sequence. Moderately moist conditions occurred during the early to middle Holocene but apparently become slightly drier after 4500 yr B.P. The impact of human settlement can be seen in the loss of woody vegetation and enhanced gullying and colluviation during the last ca. 500 years.     

High-performance liquid chromatographic determination of thiopentene enantiomers in sheep plasma

An HPLC method was developed to determine the plasma concentrations of R(+)- and S(−)-thiopentone for pharmacokinetic studies in sheep. The method required separation of the thiopentone enantiomers from the corresponding pentobarbitone enantiomers which are usually present as metabolites of thiopentone. Phenylbutazone was used as an internal standard. After acidification, the plasma samples were extracted with a mixture of ether and hexane (2:8). The solvent was evaporated to dryness and the residues were reconstituted with sodium hydroxide solution (pH 10). The samples were chromatographed on a 100 mm × 4 mm I.D.. Chiral AGP-CSP column. The mobile phase was 4.5% 2-propanol in 0.1 M phosphate buffer (pH 6.2) with a flow-rate of 0.9 ml/min. This gave k′ values of 1.92, 2.92, 5.71, 9.30 and 11.98 for R(+)-pentobarbitone, S(−)-pentobarbitone, R(+)-thiopentone, S(−)-thiopentone, and phenylbutazone, respectively. At detection wavelength of 287 nm, the limit of quantitation was 5 ng/ml for R(+)-thiopentone and 6 ng/ml for S(−)-thiopentone. The inter-day coefficients of variation at concentrations of 0.02, 0.1 and 8 μg/ml were, respectively, 4.8, 4.4 and 3.5% for R(+)-thiopentone and, respectively, 5.0, 4.3 and 3.9% for S(−)-thiopentone (n = 6 each enantiomer). At the same concentrations, the intra-day coefficients of variation from six sets of replicates (measured over six days) were, respectively, 8.0, 8.0 and 8.8% for R(+)-thiopentene and 8.8, 7.4 and 9.6% for S(−)-thiopentone. Linearity over the standard range, 0.01–40 μg/ml, was shown by correlation coefficients> 0.998. This method has proven suitable for pharmacokinetic studies of thiopentone enantiomers after administration of rac-thiopentone in human plasma also and would be suitable for pharmacokinetic studies of the pentobarbitone eantiomers.