Degradation of the gamma-Carboxyl-Containing Diarylpropane Lignin Model Compound 3-(4'-Ethoxy-3'-Methoxyphenyl)-2-(4'-Methoxyphenyl)Propionic Acid by the Basidiomycete Phanerochaete chrysosporium

The white-rot basidiomycete Phanerochaete chrysosporium metabolized 3-(4'-ethoxy-3'-methoxyphenyl)-2-(4'-methoxyphenyl)propionic acid (V) in low-nitrogen, stationary cultures, conditions under which ligninolytic activity is expressed. The ability of several fungal mutant strains to degrade V reflected their ability to degrade [C]lignin to CO(2). 1-(4'-Ethoxy-3'-methoxyphenyl)-2-(4'-methoxyphenyl)-2- hydroxyethane (VII), anisyl alcohol, and 4-ethoxy-3-methoxybenzyl alcohol were isolated as metabolic products, indicating an initial oxidative decarboxylation of V, followed by alpha, beta cleavage of the intermediate (VII). Exogenously added VII was rapidly converted to anisyl alcohol and 4-ethoxy-3-methoxybenzyl alcohol. When the degradation of V was carried out under O(2), O was incorporated into the beta position of the diarylethane product (VII), indicating that the reaction is oxygenative.     

Complexes of vanadium(III) with L-alanine and L-aspartic acid

The equilibria of the complexation processes of V(3+) with L-alanine and L-aspartic acid in aqueous solution over a wide pH range (2-10) were studied by potentiometric and spectroscopic (UV-Vis, CD) methods. The results show that alanine forms complexes with V(3+) in the metal ion concentration range and at the ligand-to-metal ratios investigated, giving mononuclear species only. In ML(2) species, which dominate in the range pH 4-8, alanine acts as a bidendate ligand through O and N atoms. The complexation processes of V(3+) with aspartic acid are more complicated. In acidic solution (up to pH approximately 4) they are similar to those for alanine. In the higher pH region, however, there are complicated equilibria among mono- and various dinuclear species. These dinuclear species consist of carboxylic or mu-oxo bridges and differ from each other by the number of coordinated ligands and OH(-) groups. The solid phase of the V(III) complex with aspartic acid could be isolated from nonaqueous solution only. Spectroscopic (UV-Vis-IR) measurements and magnetic susceptibility data confirm the coordination of vanadium(III) by two carboxylic groups. Both V(III)-L-aspartic acid and V(III)-L-alanine complexes have a significant apoptotic effect on Hepatoma Morris 5123 cells.     

New investigations on hematoxylin, hematein, and hematein-aluminium complexes. II. Hematein-aluminium complexes and hemalum staining.

The absorption spectra of hematein-aluminium solutions have been recorded at various concentrations and pH values; the solutions were prepared using analytically pure hematein and potassium alum as aluminium source. In aqueous solution, four different hematein-aluminium complexes could be distinguished by absorption spectroscopy. In weakly acidic media we observed the violet 1:1 and 1:2 complexes HmAl (VII) and HmAl2(3) (VIII), and in strongly acidic solution the red 1:1 complex HmAl2 (IX). Whereas, in weakly alkaline solution the blue 1:1 complex HmAl0 (X) was detected. By change of the pH value the complexes were mutual interconverted. The dye complexes were characterized by their absorption spectra and molar extinction coefficients. We have stained HeLa cells with the complex solutions under different experimental conditions. In all cases the nuclear staining was intense whereas the staining of the cytoplasm was weak. The microspectra of the stained nuclei were recorded and compared with the absorption spectra of the complexes in solution. Thus it was possible to identify the bound dye species. After staining in acidic media, the cells were red to red-violet depending on the reaction conditions. The three cationic dye species VII, VIII, and IX were bound in varying amounts. After blueing in weakly acidic media or in water, only the violet dye complex VII was detected whereas, after blueing in weakly alkaline media, only the blue complex X has been observed. Enzymatic digestion experiments have shown that the dye complexes in the nuclei were bound to DNA while those in the cytoplasm and nucleoli were bound to RNA. The binding between the dye complexes and the nucleic acids is discussed.     

Cathodic electrochemiluminescence of acetonitrile, acetonitrile-1,10-phenanthroline and acetonitrile-ternary Eu(III) complexes at a gold electrode.

Cathodic electrochemiluminescence (ECL) behaviours of the acetonitrile, acetonitrile-1,10-phenanthroline (phen) and acetonitrile-ternary Eu(III) complex systems at a gold electrode were studied. One very weak cathodic ECL-2 at -3.5 V was observed in 0.1 mol/L tetrabutylammonium tetrafluoroborate (TBABF(4)) acetonitrile solution. When 10 mmol/L tetrabutylammonium peroxydisulphate [(TBA)(2)S(2)O(8)] was added to 0.1 mol/L TBABF(4) acetonitrile solution, another cathodic ECL-1 at -2.7 V appeared and the potential for ECL-2 was shifted from -3.5 to -3.1 V. Furthermore, ECL-2 intensity was enhanced about 20-fold. When 1 x 10(-4) mol/L phen was added to 0.1 mol/L TBABF(4) + 10 mmol/L (TBA)(2)S(2)O(8) acetonitrile solution, the ECL intensities of ECL-1 and ECL-2 were enhanced about 20-fold compared with those of 0.1 mol/L TBABF(4) + 10 mmol/L (TBA)(2)S(2)O(8) acetonitrile solution. The maximum emission peaks of ECL-1 and ECL-2 in the three systems mentioned above appeared at about 530 nm. The products obtained by electrolysing 0.1 mol/L TBABF(4) acetonitrile solution at -3.5 V for 20 min were analysed by Fourier Transform Infrared (FTIR) spectra and gas chromatography-mass spectrometry (GC-MS) and the emitter of ECL-1 and ECL-2 was identified as excited state polyacetonitrile. When ternary Eu(III) complexes were presented in 0.1 mol/L TBABF(4) + 10 mmol/L (TBA)(2)S(2)O(8) acetonitrile solution, another maximum emission peak with a narrow band centred at about 610 nm appeared in ECL-1 in addition to the maximum emission peaks at about 530 nm for ECL-1 and ECL-2. The emitter of ECL emission at 610 nm was identified as the excited states Eu(III)*. The mechanisms for cathodic ECL behaviours of the acetonitrile, acetonitrile-phen and acetonitrile-ternary Eu(III) complex systems at a gold electrode have been proposed. The extremely sharp emission bands for ternary Eu(III) complexes may have analytical potential.     

Reaction of bis(diethylenetriamine)cobalt(III) with formaldehyde and ammonia

Condensation reactions of three isomers of bis(diethylenetriamine)cobalt(III) with formaldehyde and ammonia were studied. The s-fac isomer gave a main product containing two molecules of the cyclic amine, 1,3,5,8-tetraazacyclodecane. It was unstable in acidic solution to give several decomposition products. The encapsulated complex in which all the nitrogens were attacked by formaldehyde could not be obtained. The main product of the u-fac isomer was a half encapsulated complex, i.e., three of the four primary amino groups reacted with formaldehyde and ammonia to produce a cobalt(III) complex of 3-(7-amino-2,5-diazaheptyl)-1,3,5,8-tetraazacyclodecane. The mer isomer gave a complex of 1,4,7,9,11,14,17-heptaazaheptadecane, together with the same reaction product as the one from the u-fac isomer.     

Cross-linking of ubiquinone cytochrome c reductase (complex III) with periodate-cleavable bifunctional reagents.

Two novel cross-linkers, disuccinimidyl tartarate (DST) and N,N'-bis(3-succinimidyloxycarbonylpropyl)tartaramide (SPT), have been synthesized. These reagents span 6 and 18 A, respectively, between functional groups and contain a vic-glycol bond which can be cleaved with periodate under mild reaction conditions. Both DST and SPT have been used to examine the near-neighbor relationships of polypeptides in ubiquinone cytochrome c reductase (complex III) from beef heart mitochondria. Among the cross-linked products resolved were pairs containing I + II, II + VI, I + V, and VI + VII. Polypeptides III and IV, a cytochrome b aproprotein, and the cytochrome c1 hemoprotein, respectively, were also resolved in several cross-linked products.     

Direct and Fe(II)-mediated reduction of technetium by Fe(III)-reducing bacteria

The dissimilatory Fe(III)-reducing bacterium Geobacter sulfurreducens reduced and precipitated Tc(VII) by two mechanisms. Washed cell suspensions coupled the oxidation of hydrogen to enzymatic reduction of Tc(VII) to Tc(IV), leading to the precipitation of TcO(2) at the periphery of the cell. An indirect, Fe(II)-mediated mechanism was also identified. Acetate, although not utilized efficiently as an electron donor for direct cell-mediated reduction of technetium, supported the reduction of Fe(III), and the Fe(II) formed was able to transfer electrons abiotically to Tc(VII). Tc(VII) reduction was comparatively inefficient via this indirect mechanism when soluble Fe(III) citrate was supplied to the cultures but was enhanced in the presence of solid Fe(III) oxide. The rate of Tc(VII) reduction was optimal, however, when Fe(III) oxide reduction was stimulated by the addition of the humic analog and electron shuttle anthaquinone-2,6-disulfonate, leading to the rapid formation of the Fe(II)-bearing mineral magnetite. Under these conditions, Tc(VII) was reduced and precipitated abiotically on the nanocrystals of biogenic magnetite as TcO(2) and was removed from solution to concentrations below the limit of detection by scintillation counting. Cultures of Fe(III)-reducing bacteria enriched from radionuclide-contaminated sediment using Fe(III) oxide as an electron acceptor in the presence of 25 microM Tc(VII) contained a single Geobacter sp. detected by 16S ribosomal DNA analysis and were also able to reduce and precipitate the radionuclide via biogenic magnetite. Fe(III) reduction was stimulated in aquifer material, resulting in the formation of Fe(II)-containing minerals that were able to reduce and precipitate Tc(VII). These results suggest that Fe(III)-reducing bacteria may play an important role in immobilizing technetium in sediments via direct and indirect mechanisms.     

Studies on heterocyclic compounds: spiro [indole-3,2'-thiazolidine] derivatives. Antimicrobial activity of monohalogenated 3'-phenylspiro 3H-indole-3,2'-thiazolidine-2,4' (1H)-diones

The following halogenated 3'-phenyl [3H-indole-3,2'-thiazolidine]-2,4'(1H)-dione of general formula (A) were synthesized and screened for antimicrobial activity. (formula: see text) where: X = H (I, III, V, VII, IX, XI, XIII, XV), CH3 (II, IV, VI, VIII, X, XII, XIV, XVI); Y = H (I, II), 3-F (III, IV), 2-Cl (V, VI), 3-Cl (VII, VIII), 4-Cl (IX, X), 2-Br (XI, XII), 3-Br (XIII, XIV), 4-Br (XV, XVI). The synthetic approach involves the preparation of variously substituted Schiff-bases of indol-2,3-dione, which then are subjected to cyclocondensation with alpha-mercaptoalkanoic acids, to give spirothiazolidinones of type (A). The prepared compounds were screened against S. aureus, B. cereus, M. paratuberculosis, E. coli, S. typhi, Pr. mirabilis, Ps. aeruginosa, C. albicans, S. cerevisiae, A. niger by a disk-diffusion assay (Kirby-Bauer modified. The results of the antimicrobial screening showed that the prepared compounds exhibited varying degrees of activity against Gram-positive, Gram-negative bacteria, and fungi. 3-Fluoro-derivative (III) showed inhibitory activity especially toward S. aureus and C. albicans. Chloroderivatives (VII) and (VIII) showed broad-spectrum "in vitro" antimicrobial activity, and were especially inhibitory toward S. aureus, E. coli, and S. Typhi. Fluoro-derivative (IV) and bromo-derivatives (XIII) and (XIV) possessed marked antimicrobial activity against M. paratuberculosis.     

Novel fucolipids accumulating in human adenocarcinoma. II. Selective isolation of hybridoma antibodies that differentially recognize mono-, di-, and trifucosylated type 2 chain

A series of glycolipids having the X determinant (Gal beta 1----4 [Fuc alpha----3]GlcNAc) at the terminus and a fucosyl alpha 1----3 residue at the internal GlcNAc residue have been isolated and characterized from tumor tissues (Hakomori, S., Nudelman, E., Levery, S.B., and Kannagi, R. (1984) J. Biol. Chem. 259, 4672-4680. A series of monoclonal antibodies that differentially recognize glycolipids with mono-, di-, and trifucosylated type 2 chain have been isolated and characterized. The antibody FH4 shows a remarkable preferential reactivity towards di-/or trifucosylated type 2 chain, i.e. it does not react with monofucosylated structures, including lactofucopentaosyl (III) ceramide (III3FucnLc4), monofucosyl neolactonorhexaosylceramide (y2, V3FucnLc6), and monofucosyl neolactonoroctaosylceramide (Z1, VII3FucnLc8), but reacts well with di- and trifucosylated type 2 chain structures such as difucosyl neolactonorhexaosylceramide (III3V3Fuc2nLc6) and trifucosyl neolactonoroctaosylceramide (III3V3VII3Fuc3nLc8). Two other monoclonal antibodies, FH5 and ACFH18, preferentially react with trifucosylated type 2 chain structure (III3V3VII3Fuc3nLc8), although cross-reactivity with difucosylated type 2 chain (III3V3Fuc2nLc6) was observed. They showed a minimal cross-reaction with monofucosylated type 2 chain. In contrast, the antibody FH1 does not react with III3FucnLc4 but reacts with V3FucnLc6, III3V3Fuc2nLc6, and III3V3VII3Fuc3nLc8. Two monoclonal antibodies, FH2 and FH3, do not discriminate among various glycolipids having fucosylated type 2 chain, and their reactivities are essentially similar to previously established antibodies directed to the X determinant, such as anti-SSEA-1, WGHS 29, VEP8 and 9, My-1, etc. This series of antibodies will be useful to detect the specific type of glycolipid with fucosylated type 2 chain accumulating in human cancer and in undifferentiated cells.     

Nonmetabolic Reduction of Cr(VI) by Bacterial Surfaces Under Nutrient-Absent Conditions

We have measured the ability of nonmetabolizing cells of the bacterial species Bacillus subtilis, Sporosarcina ureae , and Shewanella putrefaciens to reduce aqueous Cr(VI) to Cr(III) in the absence of externally supplied electron donors. Each species can remove significant amounts of Cr(VI) from solution, and the Cr(VI) reduction rate is strongly dependent on solution pH. The fastest reduction rates occur under acidic conditions, with decreasing rates with increasing pH. XANES data demonstrate that Cr(VI) reduction to Cr(III) occurs within the experimental systems. Control experiments indicate that the Cr removal is not a purely adsorptive process. Reduction appears to occur at the cell wall, and is not coupled to the oxidation of bacterial organic exudates. Detailed kinetic data suggest that the reduction involves at least a two-stage process, involving an initial rapid removal mechanism followed by a slower process that follows first-order reaction kinetics. Due to the prevalence of nonmetabolizing cells and cell wall fragments in soils and deeper geologic environments, our results suggest that the observed nonmetabolic reduction of Cr(VI) to Cr(III) may significantly affect the environmental distribution of Cr in bacteria-bearing systems.     

Reduction of chromium(VI) in Chinese hamster V-79 cells

The cellular reduction of chromate(VI) was studied by electron spin resonance spectrometry. Incubation of Chinese hamster V-79 cells with Na2CrO4 resulted in the formation of both chromium(V) and chromium(III) complex in a manner dependent on time (30 min-2 h) and concentration (50-500 microM). Following removal of extracellular chromate, the level of chromium(V) complex decreased quickly during the first hour but more slowly for the next hour, whereas the level of chromium(III) remained unchanged, indicating that chromium(III) is the ultimate ion of this metal in cells. Alkaline elution studies demonstrated that treatment of cells with Na2CrO4 induced DNA single-strand breaks that decreased quickly and DNA-protein crosslinks that persisted for 2 h after removal of this metal. These results suggest that the cellular levels of chromium(V) and chromium(III) may be associated with the formation of DNA damage induced by chromium (VI).     

Direct and Fe(II)-Mediated Reduction of Technetium by Fe(III)-Reducing Bacteria

The dissimilatory Fe(III)-reducing bacterium Geobacter sulfurreducens reduced and precipitated Tc(VII) by two mechanisms. Washed cell suspensions coupled the oxidation of hydrogen to enzymatic reduction of Tc(VII) to Tc(IV), leading to the precipitation of TcO₂ at the periphery of the cell. An indirect, Fe(II)-mediated mechanism was also identified. Acetate, although not utilized efficiently as an electron donor for direct cell-mediated reduction of technetium, supported the reduction of Fe(III), and the Fe(II) formed was able to transfer electrons abiotically to Tc(VII). Tc(VII) reduction was comparatively inefficient via this indirect mechanism when soluble Fe(III) citrate was supplied to the cultures but was enhanced in the presence of solid Fe(III) oxide. The rate of Tc(VII) reduction was optimal, however, when Fe(III) oxide reduction was stimulated by the addition of the humic analog and electron shuttle anthaquinone-2,6-disulfonate, leading to the rapid formation of the Fe(II)-bearing mineral magnetite. Under these conditions, Tc(VII) was reduced and precipitated abiotically on the nanocrystals of biogenic magnetite as TcO₂ and was removed from solution to concentrations below the limit of detection by scintillation counting. Cultures of Fe(III)-reducing bacteria enriched from radionuclide-contaminated sediment using Fe(III) oxide as an electron acceptor in the presence of 25 μM Tc(VII) contained a single Geobacter sp. detected by 16S ribosomal DNA analysis and were also able to reduce and precipitate the radionuclide via biogenic magnetite. Fe(III) reduction was stimulated in aquifer material, resulting in the formation of Fe(II)-containing minerals that were able to reduce and precipitate Tc(VII). These results suggest that Fe(III)-reducing bacteria may play an important role in immobilizing technetium in sediments via direct and indirect mechanisms.     

Reduction of chromium(VI) by D-galacturonic acid and formation of stable chromium(V) intermediates

The interaction of dichromate with D-galacturonic acid in aqueous solution, as a function of pH, is described. The reaction involves the reduction of Cr(VI) to Cr(III), but the reaction rate is remarkably dependent on pH. In fact, the reduction of Cr(VI) to Cr(III) proceeds rather quickly in strongly acidic solutions, while it is slow in neutral or moderately acidic media. In all cases, according to the ESR evidence, Cr(V) species are found as intermediates. The stability of the Cr(V) species increases with increasing pH, so that it may be suggested that the overall reaction rate is controlled by the Cr(V) to Cr(III) conversion.     

Degradation of the γ-Carboxyl-Containing Diarylpropane Lignin Model Compound 3-(4′-Ethoxy-3′-Methoxyphenyl)-2-(4″-Methoxyphenyl)Propionic Acid by the Basidiomycete Phanerochaete chrysosporium

The white-rot basidiomycete Phanerochaete chrysosporium metabolized 3-(4′-ethoxy-3′-methoxyphenyl)-2-(4″-methoxyphenyl)propionic acid (V) in low-nitrogen, stationary cultures, conditions under which ligninolytic activity is expressed. The ability of several fungal mutant strains to degrade V reflected their ability to degrade [¹⁴C]lignin to ¹⁴CO₂. 1-(4′-Ethoxy-3′-methoxyphenyl)-2-(4″-methoxyphenyl)-2- hydroxyethane (VII), anisyl alcohol, and 4-ethoxy-3-methoxybenzyl alcohol were isolated as metabolic products, indicating an initial oxidative decarboxylation of V, followed by α, β cleavage of the intermediate (VII). Exogenously added VII was rapidly converted to anisyl alcohol and 4-ethoxy-3-methoxybenzyl alcohol. When the degradation of V was carried out under ¹⁸O₂, ¹⁸O was incorporated into the β position of the diarylethane product (VII), indicating that the reaction is oxygenative.     

Silent circulation of St. Louis encephalitis virus prior to an encephalitis outbreak in Cordoba, Argentina (2005)

St. Louis encephalitis virus is a complex zoonoses. In 2005, 47 laboratory-confirmed and probable clinical cases of SLEV infection were reported in Córdoba, Argentina. Although the causes of 2005 outbreak remain unknown, they might be related not only to virological factors, but also to ecological and environmental conditions. We hypothesized that one of the factors for SLE reemergence in Córdoba, Argentina, was the introduction of a new SLEV genotype (SLEV genotype III), with no previous activity in the area. In order to evaluate this hypothesis we carried out a molecular characterization of SLEV detections from mosquitoes collected between 2001 and 2004 in Córdoba city. A total of 315 mosquito pools (11,002 individuals) including 12 mosquitoes species were analyzed. Overall, 20 pools (8 mosquitoes species) were positive for SLEV. During this study, genotypes II, V and VII were detected. No mosquito pool infected with genotype III was detected before the 2005 outbreak. Genotype V was found every year and in the 8 sampled sites. Genotypes II and VII showed limited temporal and spatial activities. We cannot dismiss the association of genotype II and V as etiological agents during the outbreak. However, the silent circulation of other SLEV strains in Córdoba city before the 2005 outbreak suggests that the introduction of genotype III was an important factor associated to this event. Not mutually exclusive, other factors such as changes in avian hosts and mosquitoes vectors communities, driven by climatic and environmental modifications, should also be taken into consideration in further studies.     

Iron(III) complexation of Desferrioxamine B encapsulated in apoferritin

The dissociation of apoferritin into subunits at pH 2 followed by its reformation at pH 7.4 in presence of Desferrioxamine B (DFO) gives rise to a solution containing three DFO molecules trapped within the apoferritin (Apo-ferritin:DFO) and DFO molecules outside it. The untrapped DFO molecules in the solution were removed from Apo-ferritin:DFO by exhaustive dialysis until a negligible concentration was confirmed. The addition of Fe(III) to the dialyzed solution of Apo-ferritin:DFO resulted in the appearance of an orange-red color. The UV-Vis spectrum of this solution shows the characteristic absorption of the [DFOFe] complex centered at 425 nm. Following a similar procedure as for DFO, only one molecule of [DFOFe] was trapped in the apoferritin. The above results demonstrate the possibility of encapsulating a large molecule such as DFO in the apoferritin and, more interestingly, the ability of these DFO-encapsulated molecules to react with Fe(III) to give rise to an encapsulated [DFOFe] complex within the apoferritin.     

The effect of V(III)-adenine complex on yeast as a model of eukaryotic cells

The dinuclear micro-okso vanadium (III) complex compound H(4)V(2)OCl(4)(Ad)(2) synthesized in our laboratory was investigated as a potential cytotoxic agent against yeast cells. The results of these studies could be helpful in the explanation of the mechanism governing the V (III) compound action on yeast as a simple model of eukaryotic cells. The important factors influencing the toxicity of this complex compound are: the stage of the yeast life cycle, the rate of growth and the pH of reaction mixture. The lethal effect was distinctly stronger when the reaction mixture was slightly acidic (pH = 4). In such solutions V(III) mononuclear species with adenine was relatively stable, and during the time of experiment possibly only a slow oxidation process to V(IV) occurred. In the solutions with pH = 7, several hydrolytic, perhaps mixed-valence, polynuclear species were present and their action on the yeast cells was rather weak. The increased lethal activity of this compound in acidic solutions may be useful in specific treatment against cancer cells whose cytoplasm and/or closest surrounding has lower pH value. The next important result was an observation that the killing activity of this compound was enhanced for yeast cells being in log phase. Also these which had a slower rate of growth (possessing some auxotrophic mutations) were more resistant than those growing faster. The extent of yeast mutagenesis caused by the complex compound is negligible, as the number of mutants found in experiments was within the limit of experimental error. These results are promising and the investigated complex can be considered as a potential anti cancer agent.     

Oxidation of phenolic arylglycerol beta-aryl ether lignin model compounds by manganese peroxidase from Phanerochaete chrysosporium: oxidative cleavage of an alpha-carbonyl model compound.

Manganese peroxidase (MnP) oxidized 1-(3,5-dimethoxy-4-hydroxyphenyl)-2-(4-(hydroxymethyl)-2-methoxyphenoxy) -1,3-dihydroxypropane (I) in the presence of MnII and H2O2 to yield 1-(3,5-dimethoxy-4-hydroxyphenyl)- 2-(4-(hydroxymethyl)-2-methoxyphenoxy)-1-oxo-3-hydroxypropane (II), 2,6-dimethoxy-1,4-benzoquinone (III), 2,6-dimethoxy-1,4-dihydroxybenzene (IV), 2-(4-(hydroxymethyl)-2-methoxyphenoxy)-3-hydroxypropanal (V), syringaldehyde (VI), vanillyl alcohol (VII), and vanillin (VIII). MnP oxidized II to yield 2,6-dimethoxy-1,4-benzoquinone (III), 2,6-dimethoxy-1,4-dihydroxybenzene (IV), vanillyl alcohol (VII), vanillin (VIII), syringic acid (IX), and 2-(4-(hydroxymethyl)-2-methoxyphenoxy)-3-hydroxypropanoic acid (X). A chemically prepared MnIII-malonate complex catalyzed the same reactions. Oxidation of I and II in H2(18)O under argon resulted in incorporation of one atom of 18O into the quinone III and into the hydroquinone IV. Incorporation of one atom of oxygen from H2(18)O into syringic acid (IX) and the phenoxypropanoic acid X was also observed in the oxidation of II. These results are explained by mechanisms involving the initial one-electron oxidation of I or II by enzyme-generated MnIII to produce a phenoxy radical. This intermediate is further oxidized by MnIII to a cyclohexadienyl cation. Loss of a proton, followed by rearrangement of the quinone methide intermediate, yields the C alpha-oxo dimer II as the major product from substrate I. Alternatively, cyclohexadienyl cations are attacked by water. Subsequent alkyl-phenyl cleavage yields the hydroquinone IV and the phenoxypropanal V from I, and IV and the phenoxypropanoic acid X from II, respectively. The initial phenoxy radical also can undergo C alpha-C beta bond cleavage, yielding syringaldehyde (VI) and a C6-C2-ether radical from I and syringic acid (IX) and the same C6-C2-ether radical from II. The C6-C2-ether radical is scavenged by O2 or further oxidized by MnIII, subsequently leading to release of vanillyl alcohol (VII). VII and IV are oxidized to vanillin (VIII) and the quinone III, respectively.     

Preparation and pharmacokinetics of samarium(III)-153-labeled DTPA-bis-biotin. Characterization and theoretical studies of the samarium(III)-152 conjugate.

The complex(153)Sm(III)DTPA-bis-biotin was prepared with a 99% radiochemical purity and a specific activity of 370 MBq/mg employing a molar ratio of DTPA-bis-biotin/Sm from 2 to 4 at pH 8.0. In vitro studies demonstrated that the complex is stable after dilution in saline and in human serum. Avidity of labeled biotin for avidin was not affected by the labeling procedure. Pharmacokinetic data of (153)Sm(III)DTPA-bis-biotin in normal mice showed that blood clearance is biexponential during the time interval from 0 to 24 h and that 3 h postinjection 92 +/- 4.32% of the dose is eliminated in the urine. To have further evidence which could sustain that (153)Sm(III)DTPA-bis-biotin is stable in solution as a real coordination complex, (152)Sm(III)DTPA-bis-biotin was obtained in macroscopic quantities and its characterization was done by IR, TGA, and conductivity measurements. The results indicated that the complex was chemically pure, where the Sm(3+) ion is neutralized by three carboxylate groups of the DTPA-bis-biotin ligand and coordinated to it. Using the Force Field method followed by ab initio calculations, the DTPA-bis-biotin and the Sm(III)DTPA-bis-biotin molecules were done. Accordingly, the coordination sphere of Sm(III) was totally satisfied with nitrogen and oxygen donors; the best coordination number was 9. The conformation geometry of both compounds is presented.     

Nearest neighbor relationships of the polypeptides in ubiquinone cytochrome c reductase (complex III).

Ubiquinone cytochrome c reductase (complex III) in detergent dispersion has been cross-linked with two reversible cross-linking agents dithiobissuccinimidylpropionate and dimethyl-3.3'-dithiobispropionimidate and the cross-linked products formed have been analyzed by two-dimensional gel electrophoresis. Under mild reaction conditions, polypeptides I and II, II and VI, I and V, and VI and VII were the most prominent subunit pairs seen. With higher levels of reagent, larger aggregates were produced until an aggregate of apparent molecular weight 310 000 was the dominant band on gels. This is the complex III monomer.