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Bacterial contamination of Mediterranean coastal seawater as affected by riverine inputs: simulation approach applied to a shellfish breeding area (Thau lagoon, France).

Consequences of short-term changes in thermotolerant coliform loads on their spatio-temporal distribution in a Mediterranean lagoon with large-scale mollusk farming (Thau lagoon, France) were explored using a simulation approach. Simulations were based on bacterial transport and survival coupled models forced by the input of bacterial loads from the two main rivers (Vène and Pallas) that flow into the lagoon. Different flow types (reference, sudden and constant), bringing the same bacterial load, were considered and subsequent spatial and temporal bacterial contamination of lagoon surface water and shellfish was estimated. Simulation results showed that as long as loads were high, hydrodynamical processes governed the distribution of bacterial abundance in receiving areas. As soon as loads decreased or when time supply increased, biological die-off processes became dominant. Bacterial contamination of shellfish induced by the different flow types appeared to depend on the receiving area. In the case of Pallas River area, a sudden input of bacteria led to a high bacterial contamination of shellfish but only during a short period ( approximately 1 day). A constant input of the same amount of bacteria induced a lower but significant contamination during all the simulation period (10 days). On the contrary, bacterial inputs from the Vène River led to shellfish contamination only when bacteria were delivered through a flood event. Exposure time of bacteria to adverse environmental conditions appeared to be the main explanation to the above-mentioned differences. Consequences of our results in terms of environmental management strategy were discussed.

Animals↗

Minimum entropy production in photosynthesis.

In this paper a flux-coupling model of photosynthesis is presented. By requiring minimum entropy production, it is found that the photosynthetic efficiency is essentially given by the square root of D/lambda. D and lambda are the diffusion coefficient and thermal conductivity of the rate-limiting processes in the chloroplast, respectively. For experimental values of D and lambda, the efficiency is found to be 2.4-7.5%, with a likely value of 6.1%, whereas C4-plants are known to have an efficiency of 6.2%. We conclude that the process of photosynthesis is in quantitative agreement with the principle of minimum entropy production.

Diffusion↗

Modeling of nuclide release from low-level radioactive paraffin waste: a comparison of simulated and real waste.

Nuclide leaching models based on mass transfer theory are reviewed and evaluated to analyze the leaching test results of simulated and real paraffin waste from Korean nuclear power plants (NPPs). An empirical model (EM), bulk diffusion model (BDM), coupled diffusion/dissolution model (CDDM), shrinking core model (SCM), modified SCM (MSCM), and uniform reaction model (URM) are selected for comparison. In case of simulated paraffin waste form, the experimental results are satisfactorily explained by the SCM which is based on a diffusion-controlled dissolution reaction. Leaching behavior of real paraffin waste form is well predicted by URM that considers inter-aggregated porous medium and intra-aggregated porous medium separately. If real paraffin waste forms are manufactured with relatively uniform composition, their leaching behaviors are expected to be similar to those of simulated paraffin waste forms.

Diffusion↗

Stage-independent splicing of transcripts two heterogeneous neighboring genes in Leishmania amazonensis.

Gene expression in trypanosomatid protozoa is largely regulated posttranscriptionally, e.g., 5' splice leader addition and 3' polyadenylation of mRNAs. We examined these events in Leishmania by mapping the splice sites of the transcripts from two different, but closely linked single-copy genes 2.3 kb apart. The coding regions of the approx. 1 kb upstream gene (P36) and the approx. 1.4 kb downstream gene (NAGT) produce approx. 2 and 3 kb mRNAs, respectively. Both genes were overexpressed in cells that were transfected with this bicistronic unit (> or = 7.5 kb), taking advantage of the NAGT as a selectable marker for tunicamycin-resistance. The transcripts from both genes were spliced constitutively at both ends, irrespective of their episomal or chromosomal expression in both leishmanial stages. Primer extension of the 5' UTRs and S1 nuclease protection of the 3' UTRs initially identified the major splice sites, corresponding to the genomic sequence at -205 bp and + approx. 900 bp of P36, and -1012 bp and + approx. 600 bp of NAGT. These splice sites, consistent with the size of the major transcripts, are among those mapped precisely by sequencing RT-PCR amplified 5' and 3' UTRs. The additional sites mapped by the latter are minor alternatives, especially abundant for transcripts of the downstream NAGT. All these minor splice sites are closer than the major splice sites to the coding region, indicating that the most distant splice sites are preferentially used. This preference creates a 387 bp 'gap' with polypyrimidine tracts in the intergenic region consistent with the model coupling splice leader addition with polyadenylation in pre-mRNA processing. The stage-independence of these events suggests that the 7.5 kb dicistronic unit is suitable for constructing Leishmania-specific constitutive expression vectors.

Animals↗

Fluid flow in porous systems.

Nuclear magnetic resonance (NMR) measurements of water velocity flowing through glass bead packs with a bead diameter of 10 mm have been made using the phi echo-planar imaging (PEPI) sequence. These results indicate that for various flow rates the flow variance is proportional to the mean flow velocity in agreement with the Mansfield-Issa equation. The velocity distributions are approximately Gaussian. Investigation of the slopes of the variance vs. velocity curves as a function of slice thickness indicate some coherence effects in the connectivity of the glass bead system. An extension of an earlier intervoxel coupling model is presented, which seems to explain the observed coherence effects.

Echo-Planar Imaging↗

Transport of butyrate across the isolated bovine rumen epithelium--interaction with sodium, chloride and bicarbonate.

The Ussing chamber technique was used for studying unidirectional fluxes of 14C-butyrate across the bovine rumen epithelium in vitro. Significant amounts of butyrate were absorbed across the bovine rumen epithelium in vitro, without any external driving force. The paracellular pathway was quantitatively insignificant. The transcellular pathway was predominately voltage-insensitive. The serosal to mucosal (SM) pathway was regulated by mass action, whereas the mucosal to serosal (MS) pathway further includes a saturable process, which accounted for 30 to 55% of the MS flux. The studied transport process for 14C-butyrate across the epithelium could include metabolic processes and transport of 14C-labelled butyrate metabolites. The transport of butyrate interacted with Na+, Cl- and HCO3-, and there was a linear relationship between butyrate and sodium net transport. Lowering the sodium concentration from 140 to 10 mmol l-1 decreased the butyrate MS flux significantly. Amiloride (1 mmol l-1) did, however, not reduce the butyrate flux significantly. Chloride concentration in itself did not seem to influence the transport of butyrate, but chloride-free conditions tended to increase the MS and SM flux of butyrate by a DIDS-sensitive pathway. DIDS (bilateral 0.5 mmol l-1) did further decrease the butyrate SM flux significantly at all chloride concentrations. Removing bicarbonate from the experimental solutions decreased the MS and increased the SM flux of butyrate significantly, and abolished net butyrate flux. There were no significant effects of the carbonic anhydrase inhibitor Acetazolamide (bilateral 1.0 mmol l-1). The results can be explained by a model where butyrate and butyrate metabolites are transported both by passive diffusion and by an electroneutral anion-exchange with bicarbonate. The model couples sodium and butyrate via CO2 from metabolism of butyrate, and intracellular pH.

Animals↗

Infrared spectra of the hydrogen bond in pimelic acid crystals: polarization and temperature effects.

This paper deals with the polarized IR spectra of the hydrogen bond in pimelic acid crystals and their quantitative interpretation. The spectra were measured for the vO-H and vO-D band frequency regions, at temperatures of 298 and 77 K, for the 'alpha' crystalline form of pimelic acid. Two kinds of transmission spectra were obtained, for the beam perpendicular to the two different crystalline faces: 'ac' and 'ab'. The spectra of the hydrogen and the deuterium bonded systems were quantitatively reproduced, with sufficient accuracy, on the basis of the 'strong-coupling' model, assuming the (COOH)2 cycles to be the structural units responsible for the basic spectral properties. It was found that the spectra could be reproduced only, when assuming spectral activity in the IR of the totally symmetric proton stretching vibrations in centrosymmetric cyclic dimers of hydrogen bonds. The polarization effects in the crystal spectra were interpreted as directly connected with the hydrogen bond orientation in the crystal. However, in the pimelic acid crystalline spectra there were not observed another polarization effects characteristic for another carboxylic acid crystals, depending on differentiation of the long- and of the short-wave branch properties of each of the vO-H and vO-D bands. The temperature variation only affect the intensity ratio between the lower- and the higher-frequency branches of the vO-H and vO-D bands. These spectral effects were ascribed to less strained hydrogen bond structures in the pimelic acid lattices of the alpha' crystalline form and a relatively low concentration of defects in the lattices.

Crystallization↗

Infrared spectra of the hydrogen bond in benzoic acid crystals: temperature and polarization effects.

This paper deals with experimental studies and with quantitative interpretation of the polarized IR crystalline spectra of benzoic acid and of its deuterium isotopomers: C6H5COOD, C6D5COOH and C6D5COOD. The spectra were measured in the vO-H and in the vO-D band frequency ranges at temperatures of 298 and 77 K. The intensity distribution in the bands was quantitatively reproduced on the basis of the 'strong-coupling' model, when assuming that the isolated (COOH)2 and (COOD)2 cycles were the source of the spectral properties of the crystals. Such approach appeared to be sufficient for explaining most of the isotopic and the temperature effects in the spectra. Another band shaping mechanism, i.e. a vibronic mechanism, promoting the symmetry forbidden transition in the IR for the totally symmetric proton stretching vibrations in the centrosymmetric dimers of hydrogen bonds was also considered. A new kind of the 'long-range' isotopic effects H/D in the spectra was indicated, depending on the influence of the aromatic ring hydrogen atoms onto the vO-H band fine structure patterns. The role of the aromatic rings was also discussed, in order to explain mechanisms of extremely effective promotion of the forbidden transition, as well as of the Fermi resonance impact on the crystalline spectra.

Benzoic Acid↗

Crystal structure of a ternary complex of Tritrichomonas foetus inosine 5'-monophosphate dehydrogenase: NAD+ orients the active site loop for catalysis.

Inosine 5'-monophosphate dehydrogenase (IMPDH) catalyzes the conversion of IMP to XMP with the reduction of NAD(+), which is the rate-limiting step in the biosynthesis of guanine nucleotides. IMPDH is a promising target for chemotherapy. Microbial IMPDHs differ from mammalian enzymes in their lower affinity for inhibitors such as mycophenolic acid (MPA) and thiazole-4-carboxamide adenine dinucleotide (TAD). Part of this resistance is determined by the coupling between nicotinamide and adenosine subsites in the NAD(+) binding site that is postulated to involve an active site flap. To understand the structural basis of the drug selectivity, we solved the X-ray crystal structure of the catalytic core domain of Tritrichomonas foetus IMPDH in complex with IMP and beta-methylene-TAD at 2.2 A resolution. Unlike previous structures of this enzyme, the active site loop is ordered in this complex, and the catalytic Cys319 is 3.6 A from IMP, in the same plane as the hypoxanthine ring. The active site loop forms hydrogen bonds to the carboxamide of beta-Me-TAD which suggests that NAD(+) promotes the nucleophillic attack of Cys319 on IMP. The interactions of the adenosine end of TAD are very different from those in the human enzyme, suggesting the NAD(+) site may be an exploitable target for the design of antimicrobial drugs. In addition, a new K(+) site is observed at the subunit interface. This site is adjacent to beta-Me-TAD, consistent with the link between the K(+) activation and NAD(+). However, contrary to the coupling model, the flap does not cover the adenosine subsite and remains largely disordered.

Adenosine Diphosphate↗

Elongation properties of vaccinia virus RNA polymerase: pausing, slippage, 3' end addition, and termination site choice.

We have analyzed the elongation properties of vaccinia virus RNA polymerase during a single round of transcription in vitro. RNA-labeled ternary complexes were halted at a unique template position located upstream of a T-run (TTTTTTTTT) in the nontemplate strand; this element encodes an RNA signal for factor-dependent transcription termination at distal sites on the template. The halted ternary complexes were purified and allowed to resume elongation under a variety of conditions. We found that the T-run constituted a strong elongation block, even at high nucleotide concentrations. The principal sites of pausing were at a C position situated two nucleotides upstream of the first T in the T-run and at the first three to four T positions within the T-run. There was relatively little pausing at the five downstream Ts. Intrinsic pausing was exacerbated at suboptimal nucleotide concentrations. Ternary complexes arrested by the T-run at 10 microM NTPs rapidly traversed the T-run when the NTP pool was increased to 1 mM. Limiting GTP (1 microM) resulted in polymerase stuttering at the 3' margin of the T-run, immediately prior to a templated G position; this generated a ladder of slippage synthesis products. We found that vaccinia ternary complexes remained intact after elongating to the very end of a linear DNA template and that such complexes catalyzed the addition of extra nucleotides to the 3' end of the RNA chain. The 3' end addition required much higher concentrations of NTPs than did templated chain elongation. Finally, we report that factor-dependent transcription termination by vaccinia RNA polymerase downstream of the T-run was affected by nucleotide concentration. Limiting UTP caused the polymerase to terminate at sites closer to the UUUUUNU termination signal. This is consistent with the kinetic coupling model for factor-dependent termination.

Base Sequence↗

Vanadium, molybdenum, and sodium triethanolamine complexes derived from an assembly system containing tetrathiometalate and triethanolamine.

The reaction system composed of triethanolamine (TEA) and tetrathiovanadate in the presence of MeONa (MeOLi) or NaNH2 was studied to afford vanadium and alkali metal TEA complexes. Complexes [M subsetV6O6[N(CH2CH2O)2(CH2CH2OH)]6]2S6 (M = Na (1), Li (2)) contain two cyclic V((IV)) [12]metallacrown-6 cations linked by a polysulfide S6(2-) anion. Also separated from the reaction system is a novel sodium TEA complex [[Na(TEA)]2S6]n (3), in which parallel coordination chains [Na(TEA)+]n are connected by O-H...S hydrogen bonds forming 3-D network structure. Variable-temperature conductance of 3 was determined to display semiconductor feature. The desulfurization of VS4(3-) was observed to form S6(2-) anion and/or S8 molecule in the synthetic reactions of 1-3. A striking contrast was noted that MoS4(2-) did not desulfurize in the similar reaction to that of VS4(3-) with TEA, and a Na/TEA complex containing MoS4(2-), (Et4N)2[Na2(mu-TEA)2(CH3OH)2](MoS4)2 (4), was obtained instead. The S...H-O and O...H-O hydrogen bonds play an important role in forming the one- or three-dimensional structures for all these complexes by the linkages between clusters and chains. The IR spectrum indicates the structural similarity of complexes 1 and 2. For complex 4, the Mo=S stretch vibration at 468 cm(-1) showing widening and slight red-shift and the slight movement of the 1H NMR signals of TEA ligand to downfield are considered to be the influence of the S...H-O hydrogen bonds. Variable temperature magnetic susceptibility data were collected for complex 1. The effect magnetic moment per V6 unit varies gradually from 4.04 muB at 300 K to 6.24 muB at 5 K, exhibiting ferromagnetic interaction. Heisenberg-type vector-coupling model and molecular field approximation were used to treat the interaction between the paramagnetic sites and gave a good fitting result: J = 3.97 cm(-1), J' = 1.99 cm(-1), g = 1.99, F = 3.64 x 10(-3). Density functional calculation was also performed to complex 1 and its Li, K analogues. The lowest total bonding energy of -874.001 eV was obtained for [Na subsetV6(mu3-O)6] complex containing six unpaired electrons, indicating the stability of a V6 system with six independent spins S = 1/2. When the alkali metal ion is inserted into the [V6(mu3-O)6] cavity, the positive values of the bonding interaction (E(b)) indicate that the insertion depresses the total energy and that complexes 1 and 2 together with K+ complex may exist. However, the lowest E(b) value of the K+ complex implies its instability.

Journal Article↗

Hyperfine splittings in spin-frustrated trinuclear Cu(3) clusters.

The hyperfine structures of the EPR spectra of the spin-frustrated and distorted Cu(II) trimers were calculated in the spin-coupling model. The correlations between the hyperfine structures of the EPR spectra and geometry of the Cu(3) clusters (equilateral, isosceles, and scalene triangles) were found. For the EPR spectrum of the spin-frustrated ground state 2(S = 1/2) of an equilateral triangle Cu(3) cluster (J(12) = J(13) = J(23) = J), the calculated hyperfine structure represents the complicated spectrum of the 24 hyperfine lines, of total length 5a, where a is the hyperfine constant of the mononuclear Cu center. For an isosceles Cu(3) cluster (J(12) not equal J(13) = J(23)), the hyperfine splittings of the EPR spectra of the two split S = 1/2 levels with intermediate spins S(12) = 0 and S(12) = 1 are essentially different. The EPR signal of the |(S(12) = 0)S = 1/2> level is characterized by the four equally spaced hyperfine lines (interval A = a) with the same relative spectral amplitudes 16:16:16:16 and total length 3a. For the |(S(12) = 1)S = 1/2> level, the calculated hyperfine structure represents the spectrum of the 16 hyperfine lines with equal spacing (interval A' = a/3), the spectral intensity distribution 1:1:3:3:5:5:7:7:7:7:5:5:3:3:1:1 and total length 5a. These hyperfine spectra differ from the hyperfine structure (10 lines with interval a/3) of the EPR signals of the excited S = 3/2 level of the Cu(3) cluster. The quartet hyperfine structure, characteristic of a single Cu(2+) nucleus, which was observed experimentally for the doublet ground state of the spin-frustrated Cu(3)(II) clusters, corresponds to the hyperfine structure of the EPR signal of the |(S(12) = 0)S = 1/2> level. This hyperfine structure is evidence of the lowering of the Cu(3) cluster symmetry from trigonal to orthorhombic and the small splitting of the spin-frustrated 2(S = 1/2) ground state.

Journal Article↗

The stability and eventual lone pair distortion of the hexahalide complexes and molecules of the fifth to eighth main-group elements with one lone pair, as isolated entities or in oligomeric clusters: a vibronic coupling and DFT study.

The introduced DFT-supported vibronic coupling model together with the hardness rule indicates, for the title compounds, that the tendency toward lone pair (LP) distortions decreases with increasing coordination number (CN) and upon proceeding from fluoride to iodide as the ligands. Thus, only some hexafluoro complexes and molecules are calculated to actually undergo LP deformations; here, from the possible highest-symmetry deformations, those with C(4v) geometry possess the lowest energy, leading to the complete ablation of one ligand and, hence, explaining the nonexistence of the complexes AsF6(3-), SbF6(3-), and SF6(2-). If a lower-symmetry strain is imposed on the octahedral species, for example, induced by the simultaneous presence of terminal and bridging ligands in oligomers, the vibronic energy potential is activated. It may induce pronounced distortions, which are much larger than those of analogous clusters with central ions lacking the LP. Dimers and tetramers with common edges and faces are investigated, with the predicted and calculated deformations of the constituting octahedra again following the stability sequence C(4v) > C(2v) > C(3v). The model nicely accounts for the observed trends, as well as reproduces the experimental structures and energy balances, at least semiquantitatively; its predictive power exceeds that of the valence-shell electron-pair repulsion concept.

Journal Article↗

Macrocyclic Cu(II)(2), Cu(II)(4), Ni(II)(3), and Ni(II)(4) Complexes: Magnetic Properties of Tetranuclear Systems.

A binuclear tetraprotonated macrocyclic complex [Mg(2)(L(2)-H(4))(NO(3))(2)](NO(3))(2).6H(2)O (1) has been obtained by template condensation of 4-methyl-2,6-diformylphenol and 1,2-diaminoethane in the presence of magnesium acetate and nitrate. Complex 1 on reduction with NaBH(4), followed by the removal of magnesium, yields the 36-membered octaaminotetraphenol macrocyclic ligand H(4)L(1). The replacement of magnesium in 1 with copper(II) leads to the formation of the binuclear complex [Cu(2)L(3)(ClO(4))(2)] (2) derived from the [2+2] cyclization product of 4-methyl-2,6-diformylphenol and 1,2-diaminoethane. From H(4)L(1) a series of tetranuclear nickel(II) complexes 5-8 with the core cation [Ni(4)L(1)(&mgr;(2)-X)(2)(&mgr;(2)-H(2)O)(2)](2+) (X = NCS, N(3), OAc, or Cl) have been synthesized and characterized. The trinuclear complex [Ni(3)L(1)(acac)(2)(H(2)O)(2).2H(2)O (9), obtained by reacting nickel(II) acetylacetonate with H(4)L(1), on treatment with nickel(II) perchlorate produces the tetranuclear compound [Ni(4)L(1)(acac)(2)(H(2)O)(4)](ClO(4))(2) (10). Variable-temperature (4-300 K) magnetic susceptibility measurements have been carried out for the tetracopper(II) complex [Cu(4)L(1)(H(2)O)(4)](ClO(4))(4) (3) and the tetranickel(II) complexes [Ni(4)L(1)(&mgr;(3)-OH)(&mgr;(2)-H(2)O)(2)(ClO(4))](ClO(4))(2).2CH(3)COCH(3).H(2)O (4), [Ni(4)L(1)(&mgr;(2)-NCS)(2)(&mgr;(2)-H(2)O)(2)](ClO(4))(2).2CH(3)CN (5), [Ni(4)L(1)(&mgr;(2)-N(3))(2)(&mgr;(2)-H(2)O)(2)](ClO(4))(2).2CH(3)OH (6), [Ni(4)L(1)(&mgr;(2)-OAc)(2)(&mgr;(2)-H(2)O)(2)](ClO(4))(2).2H(2)O (7), and [Ni(4)L(1)(&mgr;(2)-Cl)(2)(&mgr;(2)-H(2)O)(2)]Cl(2).4H(2)O (8). The X-ray structure of 5 has been determined. The complex (C(50)H(70)N(12)O(14)Cl(2)S(2)Ni(4)) crystallizes in the triclinic space group P&onemacr; with a = 11.794(6) Å, b = 12.523(4) Å, c = 12.794(5) Å, alpha = 117.28(5) degrees, beta = 96.38(4) degrees, gamma = 109.65(3) degrees, and Z = 1. In the asymmetric unit each of the nickel(II) centers with distorted octahedral geometry is triply-bridged by a phenoxide group, a water molecule, and a N-bonded thiocyanate and these metal centers are further bridged to their symmetry-related counterparts by another phenoxide group. The experimental susceptibility data have been analyzed using appropriate Heisenberg spin coupling models (H = -2J(ij)()S(i)().S(j)()) and the best-fit spin exchange parameters obtained are as follows: J = -288(3) cm(-)(1) (3); J(1) = -8.1(2) cm(-)(1), J(2) = -10.2(2) cm(-)(1) (4); J(1) = -34.5(1.0) cm(-)(1), J(2) = -9.5(2.0) cm(-)(1) (5); J(1) = -34(1) cm(-)(1), J(2) = 11(2) cm(-)(1) (6); J(1) = -30(1) cm(-)(1), J(2) = -7.0(1.5) cm(-)(1) (7); J(1) = -32(1) cm(-)(1), J(2) = -4(1) cm(-)(1) (8).

Journal Article↗

Dimerization of merocyanine dyes. Structural and energetic characterization of dipolar dye aggregates and implications for nonlinear optical materials.

Aggregation of polar merocyanine dyes has been identified as an important problem in the fabrication of organic materials for photonic applications. In this work, a series of merocyanine dyes is synthesized, and their aggregation is investigated by a combination of several experimental techniques to reveal structure-property relationships. These studies provide clear evidence for the formation of centrosymmetric dimers for all investigated merocyanines in concentrated solution and in the solid state. The thermodynamics of dimerization in liquid solution is studied by concentration-dependent permittivity measurements, UV-vis spectroscopy, and electrooptical absorption experiments. A centrosymmetric dimer structure with antiparallel ordering of the dipole moments is observed in solution by 2D NMR spectroscopy as well as in the solid state by X-ray crystallography and interpreted in terms of dipolar and pi-pi interactions. The optical properties of the dimer aggregates are satisfactorily explained by an excitonic coupling model. The effect of an external electric field on the dimerization equilibrium is considered and quantitatively determined by electrooptical absorption measurements. Implications of the observed findings on the design of nonlinear optical and photorefractive materials are discussed.

Journal Article↗

A unified description of the electrochemical, charge distribution, and spectroscopic properties of the special-pair radical cation in bacterial photosynthesis.

We apply our four-state 70-vibration vibronic-coupling model for the properties of the photosynthetic special-pair radical cation to: (1) interpret the observed correlations between the midpoint potential and the distribution of spin density between the two bacteriochlorophylls for 30 mutants of Rhodobacter sphaeroides, (2) interpret the observed average intervalence hole-transfer absorption energies as a function of spin density for six mutants, and (3) simulate the recently obtained intervalence electroabsorption Stark spectrum of the wild-type reaction center. While three new parameters describing the location of the sites of mutation with respect to the special pair are required to describe the midpoint-potential data, a priori predictions are made for the transition energies and the Stark spectrum. In general, excellent predictions are made of the observed quantities, with deviations being typically of the order of twice the experimental uncertainties. A unified description of many chemical and spectroscopic properties of the bacterial reaction center is thus provided. Central to the analysis is the assumption that the perturbations made to the reaction center, either via mutations of protein residues or by application of an external electric field, act only to independently modify the oxidation potentials of the two halves of the special pair and hence the redox asymmetry E0. While this appears to be a good approximation, clear evidence is presented that effects of mutation can be more extensive than what is allowed for. A thorough set of analytical equations describing the observed properties is obtained using the Born-Oppenheimer adiabatic approximation. These equations are generally appropriate for intervalence charge-transfer problems and include, for the first time, full treatment of both symmetric and antisymmetric vibrational motions. The limits of validity of the adiabatic approach to the full nonadiabatic problem are obtained.

Journal Article↗

Sequential oxidation of the cubane [4Fe--4S] cluster from [4Fe--4S](-) to [4Fe--4S](3+) in Fe(4)S(4)L(n)(-) complexes.

Gaseous Fe(4)S(n)(-) (n = 4-6) clusters and synthetic analogue complexes, Fe(4)S(4)L(n)(-) (L = Cl, Br, I; n = 1-4), were produced by laser vaporization of a solid Fe/S target and electrospray from solution samples, respectively, and their electronic structures were probed by photoelectron spectroscopy. Low binding energy features derived from minority-spin Fe 3d electrons were clearly distinguished from S-derived bands. We showed that the electronic structure of the simplest Fe(4)S(4)(-) cubane cluster can be described by the two-layer spin-coupling model previously developed for the [4Fe] cubane analogues. The photoelectron data revealed that each extra S atom in Fe(4)S(5)(-) and Fe(4)S(6)(-) removes two minority-spin Fe 3d electrons from the [4Fe--4S] cubane core and each halogen ligand removes one Fe 3d electron from the cubane core in the Fe(4)S(4)L(n)(-) complexes, clearly revealing a behavior of sequential oxidation of the cubane over five formal oxidation states: [4Fe--4S](-) --> [4Fe--4S](0) --> [4Fe--4S](+) --> [4Fe-4S](2+) --> [4Fe-4S](3+). The current work shows the electron-storage capability of the [4Fe--4S] cubane, contributes to the understanding of its electronic structure, and further demonstrates the robustness of the cubane as a structural unit and electron-transfer center.

Anions↗

Synthesis and biological activity of the prodrug of class I major histocompatibility peptide GILGFVFTL activated by beta-glucuronidase.

The first synthesis of a prodrug of HLA-A2.1 associated antigenic influenza peptide 2a was accomplished. Two methods for synthesis of prodrugs of antigenic peptides activated by beta-glucuronidase and comprising a self-immolative 3-nitrobenzyloxycarbonyl moiety were investigated. Reaction of beta-glucuronic acid glycoside of 4-hydroxy-3-nitrobenzyl alcohol (3) with N,N'-disuccinimidyl carbonate (DSC) followed by conjugation with AlaOMe, Gly, Thr, Phe-Leu, and Leu-Arg gave carbamates 4a-4f. Deacetylation of 4b and 4e with MeONa/MeOH gave beta-glucuronides 5b and 5e. Compound 5e was converted to beta-glucuronic acid conjugate 6e by the action of pig liver esterase (PLE). Compound 6e is a substrate for beta-glucuronidase. Method of a direct introduction of the prodrug residue into antigenic nonapeptide GILGFVFTL (2b) failed. Alternately, glycine conjugate 5b was activated to pentafluorophenyl ester 10. Model coupling of 10 with Phe-Leu gave tripeptide conjugate ester 11a which was hydrolyzed by PLE to uronic acid 12. Condensation of 10 with octapeptide ILGFVFTL (9) gave prodrug precursor 11b. Octapeptide 9 was prepared by de novo synthesis using a racemization-free fragment coupling method. Ester hydrolysis with Ba(OH)(2)/MeOH gave the target prodrug 2a which is a substrate for beta-glucuronidase. Prodrug 2a does not bind to HLA-A2.1 of T2 human cells defective in major histocompatibility complex I (MHC I)-associated peptide processing. Addition of beta-glucuronidase restored the binding to the level observed with parent nonapeptide 2b although higher concentrations of prodrug 2a and enzyme were necessary.

Glucuronidase↗