Chemical models of the hydrophobic interaction: Apparent molal volumes and heat capacities of three symmetrical bolaform electrolytes and their homologous monomers in aqueous solution at 25°C

The densities and volumetric specific heats of aqueous solutions of Bu₃NHBr, Pent₃NHCl, and three diazonium salts, HN?Oct₃?NHBr₂, HN?Dec₃?NHCl₂, and Bu₃N?Oct?NBu₃Br₂, have been measured at 25°C. From these data, the apparent molal volumes φ _v and apparent molal heat capacities φ _c have been calculated and are reported here. In the series of compounds chosen, the diazonium (higher homologs) can be regarded as dimers of the alkyl-substituted ammonium ions (lower homologs), and these systems are examined as chemical models for the hydrophobic interaction. With the three homologous pairs studied here, the chemical model predicts that the strong interaction (limitingly, chemical binding) of two hydrocarbon chains in water leads to a major decrease in both φ _v and φ _c of the interacting solutes, ca.?22 cm³-mole^?1 and ?200 J-^oK^?1-mole^?1. These predictions constitute limiting behavior — useful, but not sufficient, to explain the observed concentration dependence of φ _v and φ _c in aqueous solutions of the lower homologs Bu₃NHBr, Pent₃NHCl, and Bu₄NBr. An explanation for the concentration dependence of φ _c is suggested with reference to ultrasonic relaxation data.     

In vivo monitoring of skeletal aluminum burden in patients with renal failure

In vivo neutron activation analysis was used to examine the total body and partial body (hand) aluminum levels in patients with end-stage renal failure. Patients maintained on chronic hemodialysis had higher mean body burdens of aluminum than, did those clinically managed without dialysis. Approximately 70% of the patients examined indicated elevated levels of body or skeletal aluminum. A significant correlation was observed between the in vivo aluminum/calcium ratio obtained for the hand measurement and the increase in serum aluminum levels following a disferroxamine infusion test. The direct in vivo monitoring of hand Al/Ca values in patients may provide an alternate choice to bone biopsy for the detection of aluminum intoxication.     

Spirobicyclic diamines. Part 2: Synthesis of homochiral diastereoisomeric proline derived [4.4]-spirolactams

l-Proline derived diastereoisomeric [4.4]-spirolactams have been prepared by a reductive-amination reaction of (R)- or (S)-alanine methyl ester, followed by thermal cyclisation of the resulting amine onto the proline ester group in refluxing toluene. Under similar conditions (R)- or (S)-phenylalanine methyl ester gave no cyclisation products, while R- or S-α-methylbenzylamine required treatment with NaNH₂ in refluxing toluene to induce cyclisation giving diastereoisomeric [4.4]-spirolactams.     

Mass spectrometric interrogation of thioester-bound intermediates in the initial stages of epothilone biosynthesis

Direct detection of thioester intermediate mixtures bound to EpoC, a 195 kDa polyketide synthase, has been achieved using limited proteolysis and Fourier-transform mass spectrometry (FTMS). Incubation with various N-acetylcysteamine thioester (S-NAC) substrate mimics produced mass shifts on the EpoC ACP domain consistent with their condensation with an enzyme-bound carbanion produced by the decarboxylation of methylmalonyl-S-EpoC. Reconstitution of EpoA-ACP, EpoB, and EpoC gave a +165.0 Da mass shift consistent with the formation of the methylthiazolyl-methacrylyl product by incorporation of acetyl-CoA, cysteine, and methylmalonyl-CoA. Thioester-templated reaction intermediates and products are typically characterized by quantifying radioactive substrates, either enzyme bound or chemically hydrolyzed. In contrast, the MS-based methodology described here provides semiquantifiable ratios of free enzyme, intermediate, and product occupancy and reveals that certain substrates result in a >50% formation of nonproductive intermediates.     

Reactions of 1,4-bis(tetrazole)benzenes: formation of long chain alkyl halides

The reactions of 1,4-bis[2-(tributylstannyl)tetrazol-5-yl]benzene with α,ω-dibromoalkanes were carried out in order to synthesise pendant alkyl halide derivatives of the parent bis-tetrazole. This led to the formation of several alkyl halide derivatives, substituted variously at N1 or N2 on the tetrazole ring. The crystal structures of 1,4-bis[(2-(4-bromobutyl)tetrazol-5-yl)]benzene (2-N,2-N′), 1,4-bis[(2-(4-bromobutyl)tetrazol-5-yl)]benzene (1-N,2-N′) and 1,4-bis[(2-(8-bromooctyl)tetrazol-5-yl)]benzene (2-N,2-N′) are reported. Further discussion involves the structure of 1,4-bis[2-(6-bromohexyl)-2H-tetrazol-5-yl]benzene (2-N,2-N′) previously reported.     

The scaled-particle theory and solvent isotope effects on the thermodynamic properties of inert solutes in water and methanol

The scaled-particle theory has been applied to the calculation of the thermodynamic changes associated with the formation of a cavity in several isotopic varieties of liquid water and methanol. From these results, the thermodynamic functions for the transfer of a cavity (or a hard-sphere solute) have been computed for the following solvent pairs: H₂OD₂O, H₂OH₂ ¹⁸O, H₂ ¹⁸OD₂ ¹⁸O, D₂OD₂ ¹⁸O, CH₃OHCH₃OD. For the last two of these solvents, density measurements required for the calculations were carried out as a function of temperature. The calculated deuterium solvent isotope effect on the heats and entropies of hard-sphere solutes in water is much greater than the¹⁸O isotope effect; the former also exhibits a more pronounced temperature dependence. The transfer functions computed for hard-sphere solutes are compared to experimental data on the transfer of various solutes from H₂O to D₂O and from CH₃OH to CH₃OD. In most of the cases examined, the cavity effect accounts for a large part of the transfer quantities measured for rare gases, hydrocarbons, and solutes containing a significant hydrocarbon substituent.     

Kinetic and regiospecific interrogation of covalent intermediates in the nonribosomal peptide synthesis of yersiniabactin

For interrogation of enzyme-bound intermediates in nonribosomal peptide synthetases (NRPSs), mass spectrometry is used to read out the kinetics and substrate specificity of this medicinally important class of enzymes. The protein HMWP2 (230 kDa) catalyzes 11 chemical reactions, four of which could be resolved by fast quench approaches combined with mass spectrometry. The rate of complex intermediate accumulation at the PCP1 active site was observed to occur with a rate of 19 s(-1), with the rate of cysteine acylation faster than that of intermediate translocation. Use of alternative substrates for salicylic acid (at the ArCP carrier domain) and l-cysteine (at the PCP1 carrier domain) revealed a high penalty for omission of the salicyl alcohol. For some substrates, large discrepancies were found between prior adenylation assays and the current MS-based readouts. Indirect evidence for condensation via a thiolate attack (vs an amino group) was also accumulated. This is the first report to correlate the percent occupancy of multiple active sites in parallel with kinetic and structural resolution of intermediates and provides new evidence of interdomain and intermodule communication within thiotemplate assembly lines.     

Detection of Escherichia coli O157:H7 bacteria by a combination of immunofluorescent staining and capillary electrophoresis

As the number of incidents of bacterial infections continues to rise around the globe, simpler, faster, and more sensitive diagnostic techniques are required to improve the safety of the food supply and to screen for potential bacterial infections in humans. We present here direct and indirect approaches for the detection of bacteria, which are based upon a combination of immunofluorescent staining and capillary electrophoresis. In the direct approach, Escherichia coli O157:H7 bacteria stained with fluorescein-tagged specific antibodies are detected by CE, while in the indirect approach fluorescein-tagged specific antibodies to E. coli are first captured by E. coli O157:H7 bacteria and then released and detected by CE. We have identified suitable bacteria staining and CE protocols, which involved a 10 mM Tris-borate-EDTA (TBE) buffer, 0.25 micro g antibody/1 million bacteria, and capillaries dynamically coated with poly-N-hydroxyethylacrylamide (polyDuramide). We have also successfully detected the presence of E. coli O157:H7 in contaminated meat. The total time required for analysis was 6-8 h, which is less than that realized in most commercial assays presently available.