QCM/HCC as a platform for detecting the binding of warfarin to an immobilized film of human serum albumin

Quartz crystal microbalance/heat conduction calorimetry (QCM/HCC) is a new measurement technology that has been used to monitor simultaneously the mass and motional resistance of a thin film in conjunction with the heat flow produced by a chemical change in the film initiated by reaction with a gas. In this work we examine the applicability of the QCM/HCC in detecting chemical changes at the solution/thin film interface. Human serum albumin (HSA) was bound to the gold electrode of a 5 MHz AT-cut quartz resonator using three types of linkers and then exposed to buffered solutions of the anticoagulant drug warfarin. Changes in resonator frequency and motional resistance as well as changes in heat flow produced by warfarin binding to HSA were monitored as a function of the warfarin concentration. Differences in frequency and motional resistance changes depend upon the linker and vary both in magnitude and sign, whereas the integrated heat signal is proportional to the concentration of warfarin and independent of the linker chemistry. Quartz crystal microbalance/heat conduction calorimetry can thus be a useful tool for studying protein-ligand interactions at the solution-surface interface, even though the quartz resonator does not behave as a microbalance.     

Synthesis of 17β-hydroxy-1′-H-5α-androst-2-eno[3,2-b]pyrrole and 17β-hydroxy-1′-H-5α-androst-3-eno[3,4-b]pyrrole from O-(2-hydroxyethyl)ketoxime precursors

Pyrrolosteroids such as 17β-hydroxy-1′-H-5α-androst-2-eno[3,2-b]pyrrole ( 1 ) and the novel 17β-hydroxy-1′-H-5α-androst-3-eno[3,4-b]pyrrole ( 12 ) can be synthesized from the corresponding O-(2-hydroxyethyl)ketoxime precursors. In the case of 1 , yields compare favourably with previously reported literature methods.     

PROTECTION OF ESCHERICHIA COLI CELLS AGAINST THE LETHAL EFFECTS OF ULTRAVIOLET AND X IRRADIATION BY PRIOR X IRRADIATION: A GENETIC AND PHYSIOLOGICAL STUDY

Abstract— When log phase cells of wild-type E. coli K-12 were maintained in growth medium after X irradiation, they became progressively more resistant to a subsequent exposure to UV or X radiation. The time to achieve maximum resistance was about 60 min. The uvrB, uvrD, polA and certain exrA strains (W3110 background) also demonstrated this X ray-induced resistance to subsequent UV or X irradiation but recA, recB, lex (AB1157 or W3110 backgrounds) and other exrA strains (AB1157 background) did not. The resistance induced in wild-type, uvrB and uvrD cells was characterized by the production or enhancement of a shoulder on the survival curves obtained for the second irradiation, while the resistance induced in the W3110 exrA strains was expressed only as a change in slope. The induction of resistance in the W3110 exrA strain was not inhibited by the presence of chloramphenicol, but that in the wild-type cells appeared to be. The production or enhancement of a shoulder on the survival curves of the rec ⁺ lex ⁺ exr ⁺ cells is consistent with the concept of the radiation induction of repair enzymes. Alternative explanations, however, are discussed.     

SINGLE-STRAND BREAKS IN THE DNA OF THE uvrA AND uvrB STRAINS OF ESCHERICHIA COLI K-12 AFTER ULTRAVIOLET IRRADIATION

Abstract— DNA single-strand breaks were produced in uvrA and uvrB strains of E. coli K-12 after UV (254 nm) irradiation. These breaks appear to be produced both directly by photochemical events, and by a temperature-dependent process. Cyclobutane-type pyrimidine dimers are probably not the photoproducts that lead to the temperature-dependent breaks, since photoreactivation had no detectable effect on the final yield of breaks. The DNA strand breaks appear to be repairable by a process that requires DNA polymerase I and polynucleotide ligase, but not the recA, recB, recF, lexA 101 or uvrD gene products. We hypothesize that these temperature-dependent breaks occur either as a result of breakdown of a thermolabile photoproduct, or as the initial endonucleolytic event of a uvrA , uvrB -independent excision repair process that acts on a UV photoproduct other than the cyclobutane-type pyrimidine dimer.     

Application of a simple high-performance liquid chromatographic method for the determination of melphalan in the presence of its hydrolysis products

A procedure for the separation and quantitation of melphalan (L-PAM) and its hydrolysis products by high-performance liquid chromatography is described. The hydrolysis of L-PAM at 25 +/- 0.1 degrees and 41 +/- 0.1 degrees was studied between pH 3.0 and 9.0. The pattern of hydrolysis suggested that L-PAM decomposes via two consecutive pseudo first-order reactions. Pseudo first-order rate constants (k1) were determined for the disappearance of L-PAM at various pH values in buffered solutions and in a formulated product. At both temperatures L-PAM solutions were found to be most stable at low pH. Chloride ion was found to reduce the rate of hydrolysis.     

Converged quantum calculations of HO2 bound states and resonances for J=6 and 10

Bound and resonance states of HO(2) are calculated quantum mechanically using both the Lanczos homogeneous filter diagonalization method and the real Chebyshev filter diagonalization method for nonzero total angular momentum J=6 and 10, using a parallel computing strategy. For bound states, agreement between the two methods is quite satisfactory; for resonances, while the energies are in good agreement, the widths are in general agreement. The quantum nonzero-J specific unimolecular dissociation rates for HO(2) are also calculated.     

Enantioselective synthesis of alpha-methyl-D-cysteine and lanthionine building blocks via alpha-methyl-D-serine-beta-lactone

[reaction: see text] We report here the enantioselective synthesis of Boc-alpha-methyl-d-cysteine(PMB)-OH and lanthionine building blocks through the regioselective ring opening of key intermediate Boc-alpha-methyl-d-serine-beta-lactone.     

Lignocellulose degradation in mushroom composts

The edible mushroomAgaricus bisporus is grown commercially on composted manure/straw mixtures. However, this proven composting procedure is wasteful of raw materials. A nonmanure compost was developed (Smith, 1980) with two main aims:

1. To conserve raw materials, while still producing a compost favoringAgaricus bisporus colonization and giving an economic yield of mushrooms.
2. To speed up composting, hence making more efficient use of labor, farm equipment, and buildings.

A “conservation compost” (wheat straw, bran, whey, urea, peat, and gypsum) is ready for inoculation with mushroom mycelium (spawning) after 7 d preparation, i.e., 2 d pre-wetting of straw, then 4–5 d composting under controlled conditions. Whereas a traditional manure/wheat straw compost is produced by composting in windrows (8–11 d) followed by a controlled pasteurization phase (5–7 d). In the preparation of a traditional mushroom compost, as much as 60% of the initial dry matter is lost by microbial degradation prior to spawning. By shortening the composting process to 7 d conservation of cellulose and hemicellulose is achieved with only some 30% loss in dry matter. Straw hemicelluloses are degraded much quicker than cellulose during composting. Hence, the measurable extracellular laminarinase and xylanase activities of the compost microflora appear much greater than their cellulase activities at this period in both composts. A peak in laminarinase and xylanase activity after 48 h in manure compost corresponds with the increase in microbial populations. A pronounced increase in thermophilic bacterial and actinomycete populations occurs in “conservation composts” as readily available soluble carbohydrates are assimilated. Initially, this results in higher uniform compost temperatures (60?C+) and leads to a reduced thermophilic fungal population (10³ viable propagules g^-1 dry wt compost), which may explain the lowered enzyme activities found in the “conservation composts” and thus the reduced degradation of lignocellulose. The compost microflora showed no laccase activity during composting, and little if any lignin was degraded. However,Agaricus bisporus does possess a moderately active lignolytic system and a strongly active cellulolytic system. Subsequent experiments have shown that increased mushroom yields may be obtained from these composts when urea is replaced by chicken manure as the nitrogen supplement (Smith, 1983); this has not affected compost “selectivity” for mushroom growth, dry matter loss, or the duration of the process. Although yield of mushrooms, based on compost weights at spawning tend to be lower than what would be expected from traditional composts, yield calculated on the basis of weight of starting materials is usually much higher.     

Novel mercury(II) complexes of porphyrins

Treatment of octa-alkylporphyrins (e.g. 1–3) with mercury(II) acetate in methylene chloride and tetrahydrofuran affords novel complexes (4) containing two porphyrin rings layered between three Hg atoms. A new type of stereoisomerism in regularly substituted porphyrins (e.g. 1a, 2a) is identified; it is observed by NMR spectroscopy and occurs because two forms of the complex (4) are possible, depending upon which faces of the porphyrin molecule are turned in towards the central Hg atom. The NMR spectra also indicate that in concentrated solution the double sandwich complexes (4) are stable towards disproportionation and recombination. On the basis of the inherent geometrical arrangements in the complex (4), an efficient NMR method for unambiguous identification of the four primary type isomers of the aetio- and copro-porphyrin series is described.

In very dilute solution, or in mixtures containing nucleophilic solvents such as pyridine, methanol, dioxan, or tetrahydrofuran (in the absence of excess mercury(II) acetate), the double sandwich complexes (4) are destroyed to afford normal 1:1 metalloporphyrins.