Enhanced Antiviral Function of Magnesium Chloride-Modified Heparin on a Broad Spectrum of Viruses.

Previous studies reported on the broad-spectrum antiviral function of heparin. Here we investigated the antiviral function of magnesium-modified heparin and found that modified heparin displayed a significantly enhanced antiviral function against human adenovirus (HAdV) in immortalized and primary cells. Nuclear magnetic resonance analyses revealed a conformational change of heparin when complexed with magnesium. To broadly explore this discovery, we tested the antiviral function of modified heparin against herpes simplex virus type 1 (HSV-1) and found that the replication of HSV-1 was even further decreased compared to aciclovir. Moreover, we investigated the antiviral effect against the new severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) and measured a 55-fold decreased viral load in the supernatant of infected cells associated with a 38-fold decrease in virus growth. The advantage of our modified heparin is an increased antiviral effect compared to regular heparin.

Addition of calcium and magnesium chlorides as simple means of varying bound and precipitated minerals in casein micelle: Effect on enzymatic coagulation.

In casein micelle (CM), Ca is either precipitated in the colloidal calcium phosphate (CCP) stabilized by clusters of phosphoserine (SEP) residues, or is directly bound to SEP (or glutamic and aspartic acids) of caseins without inorganic phosphate. However, it is currently not possible to titrate separately the different micellar Ca forms, making it difficult to assess their respective importance for CM properties and behavior. Both Ca2+ and Mg2+ have the same binding constants with SEP. Moreover, MgHPO4 is more soluble than CaHPO4, and its natural concentration in milk is lower. Thus, upon addition of MgCl2, Mg is mainly exchanged with CM in the bound form, whereas upon addition of CaCl2, Ca is mainly exchanged in the precipitated form. Our objective was to assess the role of the 2 forms of micellar cations (bound and precipitated) during the enzymatic coagulation of cow milk. Magnesium chloride, CaCl2, or KCl (10 mM) were added to milk and pH was adjusted to 6.6 after overnight equilibration. The KCl-supplemented milk was a positive control to assess the effect of the increased ionic strength after MgCl2 and CaCl2 addition. Mineral partition between soluble and colloidal phases after salt addition was assessed both experimentally and by using computer simulation. Enzymatic coagulation was proceeded at 30°C. Hydrolysis of κ-casein was followed by the quantitative determination of caseinomacropeptide released by RP-HPLC, aggregation of para-κ-casein micelles was measured through the evolution of backscattering properties of milk and the gel time and gel firming kinetics were determined using a Chymograph (Chr. Hansen, Horsholm, Denmark). The KCl addition did not affect mineral partition. As expected, CaCl2 addition mainly increased the CCP content, whereas the addition of MgCl2 mainly increased the bound divalent cations content. The kinetics of κ-casein hydrolysis was slowed down after CaCl2 and MgCl2 addition, and was negatively correlated with the concentration of divalent cations in the soluble phase of milk. Aggregation and gel firming curves plotted versus the progress of κ-casein hydrolysis were similar for both CaCl2- and MgCl2-supplemented milk. In view of the dual-binding model for CM assembly, this means that both Ca forms reduce electronegative repulsions between para-micelles by specific charge shielding. Inclusion of 2 Ca forms in structural models for CM allows a more detailed comprehension of how mineral equilibria affect CM properties.

Crystallization kinetics and growth of struvite crystals by seawater versus magnesium chloride as magnesium source: towards enhancing sustainability and economics of struvite crystallization.

The recycling of nutrients from wastewater and their recovery in the form of valuable products is an effective strategy to accelerate the circular economy concept. Phosphorus recovery from wastewater by struvite crystallization (MgNH4PO4·6H2O) is one of the most applied techniques to compensate for the increasing demand and to slow down the depletion rate of phosphate rocks. Using low-cost magnesium sources, such as seawater, improves the financial sustainability of struvite production. In this study, the potential of seawater for struvite crystallization versus the commonly used magnesium source, MgCl2, was tested by crystal growth and kinetic experiments. The impact of ammonium concentration, magnesium concentration and pH on the growth kinetics of struvite in synthetic and real reject water were studied. The results showed that simultaneous precipitation of calcium phosphate was insignificant when using seawater, while presence of struvite seeds diminished it further. Among the supersaturation regulators, pH had the most significant effect on the struvite growth with both MgCl2 and seawater, while high N:P molar ratios further improved the struvite crystal growth by seawater. The N:P molar ratios higher than 6 and Mg:P molar ratios higher than 0.2 are recommended to improve the crystal growth kinetics. It was concluded that seawater is a promising alternative magnesium source and the control of supersaturation regulators (i.e., Mg:P, N:P and pH) is an effective strategy to control the reaction kinetics and product properties.

The Emulsifying Properties of Hydrogenated Rosin Xylitol Ester as a Biomass Surfactant for Food: Effect of pH and Salts.

The xylitol ester of hydrogenated rosin (XEHR) was obtained for the first time from biomass-based hydrogenated rosin and xylitol using an environmentally friendly, high-pressure CO2 catalytic synthesis. This compound is intended for use as an emulsifier for food. Analyses by ICP-AES showed the absence of heavy metal residues in the product, such that it met food standards. Fourier transform infrared and nuclear magnetic resonance spectroscopies together with gel permeation chromatography confirmed the successful esterification and the formation of a monoester and diester with molar masses of 427 and 772 g/mol. The emulsification of water/soybean oil mixtures by adding the XEHR was assessed at pH values of 4, 6.86, and 10 and in the presence of NaCl, KCl, MgCl2, and CaCl2. The XEHR was found to act as an emulsifier by reducing the interfacial tension of such mixtures to less than 2 mN/m under all conditions. The highest emulsifying activity index (9.52 m2/g) and emulsifying stability index (94.53%) were obtained after adding MgCl2 (100 mM). Particle size and confocal microscopy showed that the presence of salts gave a more uniform droplet size and a finer emulsion structure. The high viscosities of the emulsions containing salts also suggested a more cohesive oil droplet network.

P-recovery in a pilot-scale struvite crystallisation reactor for source separated urine systems using seawater and magnesium chloride as magnesium sources.

Practical recovery of a non-renewable nutrient, such as phosphorus (P), is essential to support modern agriculture in the near future. The high P content of urine, makes it an attractive source for practicing the recovery of this crucial nutrient. This paper presents the experimental results at pilot-plant scale of struvite crystallisation from a source-separated urine stream using two different magnesium sources, namely magnesium chloride and seawater. The latter was chosen as sustainable option to perform P-recovery in coastal areas. Real seawater was used to assess in a more realistic way its efficiency to precipitate P as struvite, since its composition (with noticeable concentration of ions such as Ca2+, SO42-, Na+, …) could lead to the formation of impurities and other precipitates. 0.99 g of struvite was obtained per litre of urine irrespective of the operational conditions tested. In all tested conditions, precipitation efficiencies exceeded 90% and recovery efficiencies were higher than 87%, with an average struvite crystal size higher than 110 µm (and up to 320 µm, depending on the experimental conditions) in the harvested struvite samples. Almost pure struvite was obtained when MgCl2 was used as precipitant, while amorphous calcium phosphate and other impurities appeared in the precipitates using seawater as magnesium source. However, the lower settling velocity of the amorphous precipitates in comparison with the struvite precipitates suggests that their separation at industrial scale could be relatively straightforward.

Alleviating Na+ effect on phosphate and potassium recovery from synthetic urine by K-struvite crystallization using different magnesium sources.

Human urine is characterized by high concentrations of nitrogen (N), phosphorus (P) and potassium (K), of which the P and K can be recovered as K-struvite crystals. This study first investigated the formation of Na-struvite because of the high Na+ present in the urine. From the results, the optimal pH for the Na-struvite crystallization was observed to be 12, and the rise in the Na+ concentration distinctly favored the Na-struvite formation. As magnesium needed to be added to induce the K-struvite crystallization, several magnesium sources including MgCl2, Mg sacrificial electrode and Mg(OH)2 were applied to recover P and K from synthetic urine. The findings indicated that when MgCl2 was used as the magnesium source, the K removal could be slightly enhanced by prolonging the reaction time, which would correspondingly decrease the Na concentration in the precipitates; besides, the intermittent addition of MgCl2 could noticeably improve the removal efficiency of K by 6%, but simultaneously raise the Na content in the precipitates recovered. With respect to the use of the Mg sacrificial electrode, the recovery efficiencies of the P and K from synthetic urine were close to those with the use of MgCl2. However, when Mg(OH)2 was used as the magnesium source, the recovery efficiencies of P and K achieved only roughly 50%, which was much lower than those noted when MgCl2 and the Mg sacrificial electrode were employed. A comprehensive analysis revealed that the MgCl2 was the best magnesium source for the K-struvite crystallization, followed by the Mg sacrificial electrode and Mg(OH)2.

Time-Resolved Small-Angle X-ray Scattering Reveals Millisecond Transitions of a DNA Origami Switch.

Self-assembled DNA structures enable creation of specific shapes at the nanometer-micrometer scale with molecular resolution. The construction of functional DNA assemblies will likely require dynamic structures that can undergo controllable conformational changes. DNA devices based on shape complementary stacking interactions have been demonstrated to undergo reversible conformational changes triggered by changes in ionic environment or temperature. An experimentally unexplored aspect is how quickly conformational transitions of large synthetic DNA origami structures can actually occur. Here, we use time-resolved small-angle X-ray scattering to monitor large-scale conformational transitions of a two-state DNA origami switch in free solution. We show that the DNA device switches from its open to its closed conformation upon addition of MgCl2 in milliseconds, which is close to the theoretical diffusive speed limit. In contrast, measurements of the dimerization of DNA origami bricks reveal much slower and concentration-dependent assembly kinetics. DNA brick dimerization occurs on a time scale of minutes to hours suggesting that the kinetics depend on local concentration and molecular alignment.

The Evaluation of Magnesium Chloride within a Polyethylene Glycol Formulation in a Porcine Model of Acute Spinal Cord Injury.

A porcine model of spinal cord injury (SCI) was used to evaluate the neuroprotective effects of magnesium chloride (MgCl2) within a polyethylene glycol (PEG) formulation, called "AC105" (Acorda Therapeutics Inc., Ardsley, NY). Specifically, we tested the hypothesis that AC105 would lead to greater tissue sparing at the injury site and improved behavioral outcome when delivered in a clinically realistic time window post-injury. Four hours after contusion/compression injury, Yucatan minipigs were randomized to receive a 30-min intravenous infusion of AC105, magnesium sulfate (MgSO4), or saline. Animals received 4 additional infusions of the same dose at 6-h intervals. Behavioral recovery was tested for 12 weeks using two-dimensional (2D) kinematics during weight-supported treadmill walking and the Porcine Injury Behavior Scale (PTIBS), a 10-point locomotion scale. Spinal cords were evaluated ex vivo by diffusion-weighted magnetic resonance imaging (MRI) and subjected to histological analysis. Treatment with AC105 or MgSO4 did not result in improvements in locomotor recovery on the PTIBS or in 2D kinematics on weight-supported treadmill walking. Diffusion weighted imaging (DWI) showed severe loss of tissue integrity at the impact site, with decreased fractional anisotropy and increased mean diffusivity; this was not improved with AC105 or MgSO4 treatment. Histological analysis revealed no significant increase in gray or white matter sparing with AC105 or MgSO4 treatment. Finally, AC105 did not result in higher Mg2+ levels in CSF than with the use of standard MgSO4. In summary, when testing AC105 in a porcine model of SCI, we were unable to reproduce the promising therapeutic benefits observed previously in less-severe rodent models of SCI.

Concurrent Phosphorus Recovery and Energy Generation in Mediator-Less Dual Chamber Microbial Fuel Cells: Mechanisms and Influencing Factors.

This study investigated the mechanism and key factors influencing concurrent phosphorus (P) recovery and energy generation in microbial fuel cells (MFC) during wastewater treatment. Using a mediator-less dual chamber microbial fuel cell operated for 120 days; P was shown to precipitate as struvite when ammonium and magnesium chloride solutions were added to the cathode chamber. Monitoring data for chemical oxygen demand (COD), pH, oxidation reduction potential (ORP) and aeration flow rate showed that a maximum 38% P recovery was achieved; and this corresponds to 1.5 g/L, pH > 8, -550 ± 10 mV and 50 mL/min respectively, for COD, pH(cathode), ORP and cathode aeration flow rate. More importantly, COD and aeration flow rate were shown to be the key influencing factors for the P recovery and energy generation. Results further show that the maximum P recovery corresponds to 72 mW/m² power density. However, the energy generated at maximum P recovery was not the optimum; this shows that whilst P recovery and energy generation can be concurrently achieved in a microbial fuel cell, neither can be at the optimal value.

Modulation of DNA Polymerase Noncovalent Kinetic Transitions by Divalent Cations.

Replicative DNA polymerases (DNAPs) require divalent metal cations for phosphodiester bond formation in the polymerase site and for hydrolytic editing in the exonuclease site. Me(2+) ions are intimate architectural components of each active site, where they are coordinated by a conserved set of amino acids and functional groups of the reaction substrates. Therefore Me(2+) ions can influence the noncovalent transitions that occur during each nucleotide addition cycle. Using a nanopore, transitions in individual Φ29 DNAP complexes are resolved with single-nucleotide spatial precision and sub-millisecond temporal resolution. We studied Mg(2+) and Mn(2+), which support catalysis, and Ca(2+), which supports deoxynucleoside triphosphate (dNTP) binding but not catalysis. We examined their effects on translocation, dNTP binding, and primer strand transfer between the polymerase and exonuclease sites. All three metals cause a concentration-dependent shift in the translocation equilibrium, predominantly by decreasing the forward translocation rate. Me(2+) also promotes an increase in the backward translocation rate that is dependent upon the primer terminal 3'-OH group. Me(2+) modulates the translocation rates but not their response to force, suggesting that Me(2+) does not affect the distance to the transition state of translocation. Absent Me(2+), the primer strand transfer pathway between the polymerase and exonuclease sites displays additional kinetic states not observed at >1 mm Me(2+). Complementary dNTP binding is affected by Me(2+) identity, with Ca(2+) affording the highest affinity, followed by Mn(2+), and then Mg(2+). Both Ca(2+) and Mn(2+) substantially decrease the dNTP dissociation rate relative to Mg(2+), while Ca(2+) also increases the dNTP association rate.

Phosphate adsorption ability of biochar/Mg-Al assembled nanocomposites prepared by aluminum-electrode based electro-assisted modification method with MgCl2 as electrolyte.

In this work, the textural properties and phosphate adsorption capability of modified-biochar containing Mg-Al assembled nanocomposites prepared by an effective electro-assisted modification method with MgCl2 as an electrolyte have been determined. Structure and chemical analyses of the modified-biochar showed that nano-sized stonelike or flowerlike Mg-Al assembled composites, MgO, spinel MgAl2O4, AlOOH, and Al2O3, were densely grown and uniformly dispersed on the biochar surface. The adsorption isotherm and kinetics data suggested that the biochar/Mg-Al assembled nanocomposites have an energetically heterogeneous surface and that phosphate adsorption could be controlled by multiple processes. The maximum phosphate adsorption capacity was as high as 887 mg g(-1), as fitted by the Langmuir-Freundlich model, and is the highest value ever reported. It was concluded that this novel electro-assisted modification is a very attractive method and the biochar/Mg-Al assembled nanocomposites provide an excellent adsorbent that can effectively remove phosphate from aqueous solutions.

Formal synthesis of (+)-3-epi-eupomatilone-6 and the 3,5-bis-epimer.

The formal synthesis of (+)-3-epi-eupomatilone-6 () and the 3,5-bis-epimer () has been accomplished. The key synthetic strategy involved the stereoselective construction of (3R,4S,5R)- and (3R,4S,5S)-trisubstituted γ-butyrolactones and from (2R,3R)-2,3-dimethyl-4-pentenoic acid derivative , which was readily obtained via stereoselective conjugate addition of vinylmagnesium chloride to a chiral α,ß-unsaturated N-acyl oxazolidinone (Evans' auxiliary) followed by α-methylation.

Study of the recovery of phosphorus from struvite precipitation in supernatant line from anaerobic digesters of sludge.

The goal of this work was to study the effective recovery of phosphorus from the supernatant of anaerobic digestion of sewage sludge by precipitation as struvite. The formation of struvite is envisioned as a promising process for nutrient removal and subsequent recovery, thus providing a strong incentive for its implementation, since the sewage is a renewable source of phosphorus. Struvite precipitation was obtained by controlled addition of Mg(OH)2 or MgCl2. We evaluated the removal of ammonia and phosphate under equimolar conditions of magnesium and magnesium stoichiometric excess of 100 to 200% relative to the limiting reagent, under a stirring speed of 300 rpm at pH 8, 9 and 10. The best condition was MgCl2 in 1:1 molar ratio to phosphate, considering the stoichiometric ratio [PO4(3-)]:[NH4(+)] of 0.13 (presented by raw sample). The results show the best cost-benefit ratio, removal of phosphate of 90.6% and ammonium removal of 29%, resulting in 23 mg l(-1) PO4(3-) and 265 mg l(-1) NH4(+) concentration in effluent.

Effects of supplementation with different Mg salts in cells: is there a clue?

The differing bioavailability of magnesium salts remains an open question, both at the cellular and systemic level. However, this issue is relevant for identifying the most effective magnesium supplement. We compared the effects of three widely used magnesium salts: MgSO4, MgCl2 and Mg pidolate, on the proliferation of four human cell types: promyelocytic leukaemia HL60, osteoblast-like Saos-2 and U-2 OS, and endothelial cells from the umbilical vein. The three magnesium salts had no effect on endothelial and leukemic cell growth, but magnesium pidolate impaired cell growth in osteoblast-like cells. In particular, in Saos-2 cells, 1 mM pidolate induced a slight accumulation of cells in the G0/G1 phase of the cell cycle and, in parallel, an early rise in intracellular calcium and a late decrease in intracellular magnesium content. Interestingly, when cultured in 5 mM magnesium pidolate, Saos-2 cells grew as fast as the controls. Moreover, intracellular magnesium and calcium concentrations did not vary. These results suggest a lower bioavailability of magnesium pidolate in osteoblast-like cells.

Determination of phosphorus fertilizer soil reactions by Raman and synchrotron infrared microspectroscopy.

The reaction mechanisms of phosphate-bearing mineral phases from sewage sludge ash-based fertilizers in soil were determined by Raman and synchrotron infrared microspectroscopy. Different reaction mechanisms in wet soil were found for calcium and magnesium (pyro-) phosphates. Calcium orthophosphates were converted over time to hydroxyapatite. Conversely, different magnesium phosphates were transformed to trimagnesium phosphate. Since the magnesium phosphates are unable to form an apatite structure, the plant-available phosphorus remains in the soil, leading to better growth results observed in agricultural pot experiments. The pyrophosphates also reacted very differently. Calcium pyrophosphate is unreactive in soil. In contrast, magnesium pyrophosphate quickly formed plant-available dimagnesium phosphate.

Effect of water hardness on surface tension and dilational visco-elasticity of sodium dodecyl sulphate solutions.

The complementary drop and bubble profile analysis and maximum bubble pressure tensiometry are used to measure the dynamic surface tension of aqueous SDS solutions in the presence of hardness salts (CaCl(2) and MgCl(2) in the ratio of 2:1 at concentrations of 6 and 40FH). The presence of hardness salts results in an essential increase of the SDS adsorption activity, which indicates the formation of Ca(DS)(2) and Mg(DS)(2) in the SDS solutions. The surface tension isotherms of SDS in presence of Ca(DS)(2) and Mg(DS)(2) are described using the generalised Frumkin model. The presence of hardness salts accelerates the ageing of SDS solutions as compared with the addition of 0.01 M NaCl due to a faster hydrolysis and hence formation of dodecanol. These results are used to estimate the possible concentration of dodecanol in the studied SDS solutions. The buoyant bubble profile method with harmonic surface oscillations is used to measure the dilational rheology of SDS solutions in presence of hardness salts in the frequency range between 0.005 Hz and 0.2 Hz. The visco-elasticity modulus in the presence of hardness salts is higher as compared with its values in the presence of 0.01 M NaCl additions. The ageing of SDS solutions leads to an essential increase of the visco-elastic modulus.

Moisture-induced aggregation of alpha-lactalbumin: effects of temperature, cations, and pH.

UNLABELLED: Alpha-lactalbumin is an important dairy protein ingredient, and has been widely used in high-protein foods such as infant formula and nutritional bars for its nutritional and functional properties. The purpose of this study was to investigate the moisture-induced aggregation of alpha-lactalbumin in premixed protein dough model systems, and to illustrate the effects of temperature, cations, and pH on the progress of protein aggregation. Our results suggested that storage temperature was a critical factor for protein aggregation in model systems, and the formation of protein aggregates became faster with increases in storage temperature. Calcium significantly improved the thermal stability of alpha-lactalbumin and slowed down the formation of protein aggregates. The increases in pH accelerated the aggregation of alpha-lactalbumin. Our results also suggested that the formation of intermolecular disulfide bonds together with noncovalent interactions are the main mechanisms resulting in the moisture-induced aggregation of alpha-lactalbumin in model systems. PRACTICAL APPLICATION: Alpha-lactalbumin is an important dairy protein ingredient, and has been widely used in high-protein foods such as infant formula and nutritional bars for its nutritional and functional properties. Our results suggested low storage temperature, the presence of calcium and low pH condition can make high-protein food products containing alpha-lactalbumin more stable.

Conductivity and electrophoretic mobility of dilute ionic solutions.

Two complementary continuum theories of electrokinetic transport are examined with particular emphasis on the equivalent conductance of binary electrolytes. The "small ion" model [R.M. Fuoss, L. Onsager, J. Phys. Chem. 61 (1957) 668] and "large ion" model [R.W. O'Brien, L.R. White, J. Chem. Soc. Faraday Trans. 2 (74) (1978) 1607] are both discussed and the "large ion" model is generalized to include an ion exclusion distance and to account in a simple but approximate way for the Brownian motion of all ions present. In addition, the "large ion" model is modified to treat "slip" hydrodynamic boundary conditions in addition to the standard "stick" boundary condition. Both models are applied to the equivalent conductance of dilute KCl, MgCl(2), and LaCl(3) solutions and both are able to reproduce experimental conductances to within an accuracy of several tenths of a percent. Despite fundamental differences in the "small ion" and "large ion" theories, they both work equally well in this application. In addition, both "stick-large ion" and "slip-large ion" models are equally capable of accounting for the equivalent conductances of the three electrolyte solutions.

Quantification of copper phases, their reducibility and dispersion in doped-CuCl2/Al2O3 catalysts for ethylene oxychlorination.

The comprehensive understanding of the composition, behaviour and reactivity of a catalyst used inside industrial plants is an extremely hard task that is rarely achieved. It requires the use of different spectroscopic techniques, applied under in situ or in operando conditions, and combined with the investigation of the catalyst activity. Often the operating experimental conditions are different from technique to technique and the different results must be compared with care. In the present contribution, we combined in situ XANES/EXAFS, IR spectroscopy of adsorbed CO, CO chemisorption and catalytic tests performed using a pulse reactor in depletive mode. This multitechnical approach resulted in the understanding of the role that dopants (LiCl, KCl, CsCl, MgCl(2) LaCl(3)) have in the nature, relative fraction, reducibility and dispersion of Cu-phases on CuCl(2)/gamma-Al(2)O(3) catalysts for oxychlorination reaction, a key step of the PVC chemistry. In the undoped catalyst two Cu phases coexist: Cu-aluminate and supported CuCl(2), being the latter the only active one [J. Catal., 2000, 189, 91]. EXAFS and XANES highlighted that all dopants contribute more or less efficiently in increasing the fraction of the active copper species, that reaches a value of almost 100% in the case of MgCl(2) or LaCl(3). EXAFS directly, and IR indirectly, proved that the addition of KCl or CsCl (and less efficiently of LiCl) results in the formation of mixed CuK(x)Cl(2+x) or CuCs(x)Cl(2+x) phases, so altering the chemical nature of the active phase. XANES spectroscopy indicates that addition of MgCl(2) or LaCl(3) does not affect the reducibility by ethylene (under static conditions) of the active CuCl(2) phase and that the reducibilility of the new copper-dopant mixed chloride are in the order CuCl(2) > CuLi(x)Cl(2+x) > CuK(x)Cl(2+x) > CuCs(x)Cl(2+x). However, when reduction is done inside a pulse reactor, a more informative picture comes out. The last technique is able to differentiate all samples, and their ability to be reduced by ethylene resulted in the order: La- > Mg- > Li-doped > undoped > K- > Cs-doped catalyst. To understand this apparent discrepancy the dispersion of the active phase, measured by CO chemisorption, was needed: it has been found that addition of LiCl increases enormously the dispersion of the active phase, LaCl(3) significantly and MgCl(2) barely, while addition of both KCl and CsCl results in a decrease of the surface area of the active phase. The mechanism of the enhancing effect of La and Mg on catalytic activity is still not clear, but it could be associated to the modification that they induce to the support surface: the Cu is so highly dispersed that almost all is in direct contact with support surface. It is finally worth noticing that the previous EXAFS and XANES study allowed us to refer the chemisorption data to the active phase only, while the IR study allowed us to fix the Cu(+)/CO surface stoichiometry. Summarizing the use of a multidisciplinary approach has been the conditio sine qua non (mandatory condition) to understand the complex role that the different additives have on the active phase of the CuCl(2)/gamma-Al(2)O(3) catalysts for ethylene oxychlorination.

Solution conformation and dynamics of the HIV-1 integrase core domain.

The human immunodeficiency virus type 1 (HIV-1) integrase (IN) is a critical enzyme involved in infection. It catalyzes two reactions to integrate the viral cDNA into the host genome, 3' processing and strand transfer, but the dynamic behavior of the active site during catalysis of these two processes remains poorly characterized. NMR spectroscopy can reveal important structural details about enzyme mechanisms, but to date the IN catalytic core domain has proven resistant to such an analysis. Here, we present the first NMR studies of a soluble variant of the catalytic core domain. The NMR chemical shifts are found to corroborate structures observed in crystals, and confirm prior studies suggesting that the alpha4 helix extends toward the active site. We also observe a dramatic improvement in NMR spectra with increasing MgCl(2) concentration. This improvement suggests a structural transition not only near the active site residues but also throughout the entire molecule as IN binds Mg(2+). In particular, the stability of the core domain is linked to the conformation of its C-terminal helix, which has implications for relative domain orientation in the full-length enzyme. (15)N relaxation experiments further show that, although conformationally flexible, the catalytic loop of IN is not fully disordered in the absence of DNA. Indeed, automated chemical shift-based modeling of the active site loop reveals several stable clusters that show striking similarity to a recent crystal structure of prototype foamy virus IN bound to DNA.