THE CONTROL OF METABOLISM BY BLUE LIGHT IN Acetabularia mediterranea—II. SELECTIVE TRANSLATIONAL CONTROL AND DIFFERENTIAL DEGRADATION OF ENZYMES

Abstract— During prolonged continuous irradiation with red light the specific activity of uridine 5'-diphosphoglucose (UDPG) pyrophosphorylase (uridine 5'-triphosphate: glucose 1-phosphate uridylyl-transferase EC 2.7.7.9) decreased in Acetabularia mediterranea Lamouroux (=A. acetabulum (L.) Silva). Subsequent blue light restored the original activity within a comparatively short period of 3 to 4 days. Computer-aided quantitative evaluation of density labelling experiments showed that the synthesis of the enzyme was accelerated about four-fold during the period of activation by blue light. A similar increase in the rate of synthesis was found for hydroxypyruvate reductase (EC 1.1.1.81), a control enzyme that showed no blue light-dependent changes in the specific activity under these conditions. The increase in the rate of enzyme synthesis was caused by an overall stimulation of the cytosolic translation. Degradation of UDPG pyrophosphorylase was unaffected by blue light, while the half life of hydroxypyruvate reductase was shortened about two-fold compared to continuous red light. Thus, degradation of proteins appears to be selectively light dependent in Acetabularia.
Model calculations for enzyme amount and enzyme synthesis were carried out using the measurements of enzyme activity, rates of cytosolic protein synthesis, and degradation constants of the enzymes. Assuming that activities represented amounts of the given enzymes, these calculations indicated a selective activation of UDPG pyrophosphorylase synthesis by blue light since it did not coincide with the overall stimulation of protein synthesis in the cytosol, in contrast to hydroxypyruvate reductase.     

Ultrafast energy-electron transfer cascade in a multichromophoric light-harvesting molecular square

A molecular square with dimensions of about 4 nm, incorporating sixteen pyrene chromophores attached to four ditopic bay-functionalized perylene bisimide chromophores, has been synthesized by coordination to four Pt(II) phosphine corner units and fully characterized via NMR spectroscopy and ESI-FTICR mass spectrometry. Steady-state and time-resolved emission as well as femtosecond transient absorption studies reveal the presence of a highly efficient (>90%) and fast photoinduced energy transfer (k(en) approximately equal to 5.0 x 10(9) s(-1)) from the pyrene to the perylene bisimide chromophores and a very fast and efficient electron transfer (>94%, k(et) approximately equal to 5 x 10(11) up to 43 x 10(11) s(-1)). Spectrotemporal parametrization indicates upper excited-state electron-transfer processes, various energy and electron-transfer pathways, and chromophoric heterogeneity. Temperature-dependent time-resolved emission spectroscopy has shown that the acceptor emission lifetime increases with decreasing temperature from which an electron-transfer barrier is obtained. The extremely fast electron-transfer processes (substantially faster and more efficient than in the free ligand) that are normally only observed in solid materials, together with the closely packed structure of 20 chromophoric units, indicate that we can consider the molecular square as a monodisperse nanoaggregate: a molecularly defined ensemble of chromophores that partly behaves like a solid material.     

Bank of reference samples of blank urine from livestock

From three species of livestock, bovine, ovine and porcine, samples of urine from each 10–20 different animals were collected. The animals originated from a governmental experimental farm and differed in race, age, sex and were fed with different types of feed. Fifty different samples of urine were lyophilised in units of 5 mL in sealed amber glass bottles. After lyophilisation of the samples, several quality control tests were performed. The variation in net weight of the units of urine and the lyophilisates ranged from 0.01%–1.7% and 0.1%–10%, respectively. Most values did not exceed 0.1% for urines and 1% for lyophilisates. The residual water content of the lyophilisates was determined by the Karl-Fisher titration method. The average percentage of residual water ranged from 1.0%–7.0% for bovine, from 1.3%–3.0% for porcine and from 1.6%–5.5% for ovine urine lyophilisates. Further, all different samples were analysed for the presence of anabolic compounds with a multi-residue procedure developed at the CRL. In two samples of bovine urine the presence of α-boldenone was detected and confirmed. In most porcine urines the endogenous steroids α-testosterone and α-estradiol were detected. In a number of porcine and ovine urine samples α- and β-zearalenol were detected. The origin of these compounds is the f2-toxin produced by a Fusarium fungus. In ovine samples α-testosterone and α-estradiol were detected.     

Self-assembled monolayers of dendritic polyglycerol derivatives on gold that resist the adsorption of proteins

Highly protein-resistant, self-assembled monolayers (SAMs) of dendritic polyglycerols (PGs) on gold can easily be obtained by simple chemical modification of these readily available polymers with a surface-active disulfide linker group. Several disulfide-functionalized PGs were synthesized by N,N'-dicyclohexylcarbodiimide-mediated ester coupling of thioctic acid. Monolayers of the disulfide-functionalized PG derivatives spontaneously form on a semitransparent gold surface and effectively prevent the adsorption of proteins, as demonstrated by surface plasmon resonance (SPR) kinetic measurements. A structure-activity relationship relating the polymer architecture to its ability to effectuate protein resistance has been derived from results of different surface characterization techniques (SPR, attenuated total reflectance infrared (ATR-IR), and contact-angle measurements). Dendritic PGs combine the characteristic structural features of several highly protein-resistant surfaces: a highly flexible aliphatic polyether, hydrophilic surface groups, and a highly branched architecture. PG monolayers are as protein resistant as poly(ethylene glycol) (PEG) SAMs and are significantly better than dextran-coated surfaces, which are currently used as the background for SPR spectroscopy. Due to the higher thermal and oxidative stability of the bulk PG as compared to the PEG and the easy accessibility of these materials, dendritic polyglycerols are novel and promising candidates as surface coatings for biomedical applications.     

Determination of low-molecular-mass quaternary ammonium compounds by capillary electrophoresis and hyphenation with mass spectrometry

The determination of quaternary ammonium ions by capillary electrophoresis (CE) is reviewed. The analytes include tetraalkylammonium and alkylbenzyldimethylammonium compounds frequently used as antiseptic and antibacterial agents as well as in various household products, several plant growth regulators and herbicides, by-products in bile acid sequestrants, and a range of anticholinergic drugs. Besides direct and indirect UV detection, hyphenation with electrospray mass spectrometry is particularly suited for quaternary ammonium ions and may lower the detection limits by two orders of magnitude. In comparison with established liquid chromatographic techniques, CE may exhibits superior separation efficiency. Applications in routine analysis have demonstrated that CE is reliable and robust enough to represent a real alternative to chromatography.     

α-Methylidene- and α-Alkylidene-β-lactams from Nonproteinogenic Amino Acids

Treatment of methyl 2-(1-hydroxyalkyl)prop-2-enoates 1 with conc. HBr solution afforded methyl (Z)-2-(bromomethyl)alk-2-enoates 2 , which were transformed regioselectively into N-substituted methyl (E)-2- (aminomethyl)alk-2-enoates 3 (S_N2 reaction) and into N-substituted methyl 2-(1-aminoalkyl)prop-2-enoates 4 (S_N2′ reaction). Regiocontrol of nucleophilic attack by amine was accomplished simply by choice of solvent, the S_N2 reaction occurring in MeCN and the S_N2′ reaction in petroleum ether. Hydrolysis and lactamization afforded β-lactams 7 and 8 , containing an exocyciic alkylidene and methylidene group at C(3), respectively.     

Wavelength-dependent electron and energy transfer pathways in a side-to-face ruthenium porphyrin/perylene bisimide assembly

A new side-to-face supramolecular array of chromophores, where a pyridyl-substituted perylene bisimide dye axially binds to two ruthenium porphyrin fragments, has been prepared by self-assembly. The array is formulated as DPyPBI[Ru(TPP)(CO)](2), where DPyPBI = N,N'-di(4-pyridyl)-1,6,7,12-tetra(4-tert-butylphenoxy)perylene-3,4:9,10-tetracarboxylic acid bisimide and TPP = 5,10,15,20-tetraphenylporphyrin. The photophysical behavior of DPyPBI[Ru(TPP)(CO)](2) has been studied by fast (nanoseconds) and ultrafast (femtoseconds) time-resolved techniques. The observed behavior sharply changes with excitation wavelength, depending on whether the DPyPBI or Ru(TPP)(CO) units are excited. After DPyPBI excitation, the strong fluorescence typical of this unit is completely quenched, and time-resolved spectroscopy reveals the occurrence of photoinduced electron transfer from the ruthenium porphyrin to the perylene bisimide dye (tau = 5.6 ps) followed by charge recombination (tau = 270 ps). Upon excitation of the Ru(TPP)(CO) fragments, on the other hand, ultrafast (tau < 1 ps) intersystem crossing is followed by triplet energy transfer from the ruthenium porphyrin to the perylene bisimide dye (tau = 720 ps). The perylene-based triplet state decays to the ground state on a longer time scale (tau = 9.8 micros). The photophysics of this supramolecular array provides remarkable examples of (i) wavelength-dependent behavior (a small change in excitation wavelength causes a sharp switch from electron to energy transfer) and (ii) intramolecular sensitization (the triplet state of the perylene bisimide, inaccessible in the free dye, is efficiently populated in the array).     

Syntheses and Structure of Gd<Subscript>3</Subscript>Rh<Subscript>1.940(7)</Subscript>In<Subscript>4</Subscript>

Summary. The gadolinium–rhodium–indide Gd₃Rh_1.940(7)In₄ was prepared by arc-melting of the elements and subsequent annealing in a corundum crucible in a sealed silica tube. Gd₃Rh_1.940(7)In₄ adopts the hexagonal Lu₃Co_1.87In₄ type, space group , a = 781.4(5), c = 383.8(3) pm, wR2 = 0.0285, BASF = 0.375(1) (merohedric twinning via a twofold axis (xx0)), 648 F² values, 22 variables. The structure is derived from the well known ZrNiAl type through an ordering of rhodium and indium atoms on the Ni2 sites. The Rh/In ordering forces a reduction of the space group symmetry from to , leading to merohedric twinning for the investigated crystal. The Rh1 site has an occupancy of only 94.0(7)%. The investigated crystal had a composition Gd₃Rh_1.940(7)In₄. The main geometrical motif are three types of centered, tricapped trigonal prisms, i.e., [Rh1In2₆Gd₃], [Rh2Gd₆In2₃], and [In1Gd₆In2₃]. The shortest interatomic distances occur for Rh–In (276–296 pm) followed by In–In (297 pm). Together, the rhodium and indium atoms build up a three-dimensional [Rh_1.940(7)In₄] network, in which the gadolinium atoms fill slightly distorted pentagonal channels. The crystal chemistry of Gd₃Rh_1.940(7)In₄ is discussed on the basis of a group-subgroup scheme.     

(A19N7)[In4]2 (A = Ca,Sr) and (Ca4N)[In2]: Synthesis,Crystal Structures,Physical Properties,and Chemical Bonding

Single phase powders of (A₁₉N₇)[In₄]₂ (A = Ca, Sr) and (Ca₄N)[In₂] were prepared by reaction of melt beads of the metallic components with nitrogen. The crystal structure of (Ca₁₉N₇)[In₄]₂ was refined based on neutron and X‐ray powder diffraction data. The crystal structure of (Sr₁₉N₇)[In₄]₂ was solved from the X‐ray powder pattern. The structure refinements in combination with results from chemical analyses ascertain the compositions. The compounds (A₁₉N₇)[In₄]₂ (A = Ca, Sr) are isotypes of (Ca₁₉N₇)[Ag₄]₂; (Ca₁₉N₇)[In₄]₂ is probably identical to the earlier reported (Ca_18.5N₇)[In₄]₂. The crystal structure of the isotypes (A₁₉N₇)[In₄]₂ (A = Ca, Sr; cubic, , Ca: a = 1471.65(3) pm; Sr: a = 1561.0(1) pm) contains isolated [In₄] tetrahedra embedded in a framework of edge‐ and vertex‐sharing (A₆N) octahedra. Six of these octahedra are condensed by edge‐sharing around one central A²⁺ ion to form “superoctahedra” (A₁₉N₆) which are connected three‐dimensionally via further octahedra by corner‐sharing. The crystal structure of (Ca₄N)[In₂] (tetragonal, I4₁/amd, a = 491.14(4) pm, c = 2907.7(3) pm) consists of alternating layers of perovskite type slabs of vertex‐sharing octahedra (Ca₂Ca_4/2N) and parallel arranged infinite zigzag chains equation/tex2gif-stack-1.gif[In₂]. In the sense of Zintl‐type counting the compounds (A²⁺)₁₉(N^3?)₇[(In^2.125?)₄]₂ present an electron excess, (Ca²⁺)₄(N^3?)[(In^2.5?)₂] is electron deficient. Metallic properties are supported by electrical resistivity and magnetic susceptibility measurements. The analysis of the electronic structures gives evidence for the existence of homoatomic interactions In–In and significant heteroatomic metal–metal interactions Ca–In which favor the deviations of the title compounds from the (8 – N) rule.