Identification of hepatic metabolites of two highly carcinogenic polycyclic aza-aromatic compounds, 7,9-dimethylbenz[c]acridine and 7,10-dimethylbenz[c]acridine

The hepatic microsomal metabolites of the highly carcinogenic dimethylbenzacridines, 7,9-dimethylbenz[c]acridine (7,9-DMBAC), and 7,10-dimethylbenz[c]acridine (7,10-DMBAC) were obtained with preparations from 3-methylcholanthrene-pretreated rats. Metabolites were separated by reversed-phase HPLC and characterized using UV spectral data and chemical ionization-mass spectrometry after trimethylsilylation and GC. Comparisons with products formed in the presence of the epoxide hydrolase inhibitor, 1,1,1-trichloropropane 2,3-oxide and with those formed from the three synthetic alcohol derivatives of each parent compound, aided the assignment of firm or tentative structures to 16 products from 7,9-DMBAC found in 22 reversed-phase chromatographic peaks, and for 17 products of 7,10-DMBAC found in 19 chromatographic peaks. The more abundant metabolites were derived from oxidation of the methyl groups. Other metabolites were dihydrodiols, epoxides, phenols and secondary metabolites. The 9-methyl group prevented dihydrodiol formation at the 8,9-position from 7,9-DMBAC, and for each carcinogen, the 3,4-dihydrodiol was formed. As well, 3,4-dihydrodiols of methyl oxidized compounds were found.     

Partitioning multiple objective optimal solutions with applications in radiotherapy design

The optimal partition for linear programming is induced by any strictly complementary solution, and this partition is important because it characterizes the optimal set. However, constructing a strictly complementary solution in the presence of degeneracy was not practical until interior point algorithms became viable alternatives to the simplex algorithm. We develop analogs of the optimal partition for linear programming in the case of multiple objectives and show that these new partitions provide insight into the optimal set (both pareto optimality and lexicographic ordering are considered). Techniques to produce these optimal partitions are provided, and examples from the design of radiotherapy plans show that these new partitions are useful. A. Holder. Research conducted at Trinity University, TX, and The University of Mississippi, MS. This research was partially supported by ONR Grant N00014-01-1-0917.     

Segmental reflex sympathetic dystrophy: clinical and scintigraphic criteria

Clinical and scintigraphic criteria are proposed for the diagnosis of segmental reflex sympathetic dystrophy. Eight patients met previously described clinical criteria for reflex sympathetic dystrophy with involvement limited to only a portion of the hand. The delayed phase of the three-phase radionuclide bone scan was found to be highly sensitive (100%) for this small group of patients. Consecutive bone scans (n = 127) performed during a 6-month period for a variety of upper extremity problems were reviewed, and a segmentally diffuse pattern of tracer uptake was found to be highly specific (98%) for segmental reflex sympathetic dystrophy. Recognition and documentation of a more localized form of reflex sympathetic dystrophy will allow earlier recognition and treatment, which is an important factor in a successful outcome for managing pain dysfunction disorders.     

Development of technologies aiding large-tissue engineering

Apoptosis associated oligonucleosomal fragmentation of DNA can result from the activation of endonucleases that exhibit different pH optima and are either sensitive or insensitive to divalent cations. DNA fragmentation due to activation of cation sensitive endonucleases occurs in the absence of a change in intracellular pH whereas intracellular acidification is a feature of apoptosis characterized by activation of cation insensitive acidic endonuclease. We have reported earlier that somatostatin (SST) induced DNA fragmentation and apoptosis is signaled in a receptor subtype selective manner uniquely via human somatostatin receptor subtype 3 (hSSTR3). In the present study we investigated the pH dependence and cation sensitivity of endonuclease induced in hSSTR3 expressing CHO-K1 cells by the SST agonist octreotide (OCT) and its effect on intracellular pH. We show that OCT induced apoptosis is associated with selective stimulation of a divalent cation insensitive acidic endonuclease. The intracellular pH of of cells undergoing OCT induced apoptosis was 0.9 pH units lower than that of control cells. The effect of OCT on endonuclease and pH was inhibited by orthovanadate as well as by pretreatment with pertussis toxin, suggesting that hSSTR3 initiated cytotoxic signaling is protein tyrosine phosphatase mediated and is G protein dependent. These findings suggest that intracellular acidification and activation of acidic endonuclease mediate wild type p53 associated apoptosis signaled by hormones acting via G protein coupled receptors.     

Semiempirical and ab initio conformational analysis of 2-methylene-8,8-dimethyl-1,4,6,10-tetraoxaspiro[4.5] decane with application of GIAO-SCF methods to NMR spectrum interpretation

The GIAO-SCF method for calculating isotropic nuclear magnetic shielding values has been utilized to explain certain features in the ¹H-NMR spectrum of 2-methylene-8,8-dimethyl-1,4,6,10-tetraoxaspiro[4.5] decane. Population distributions of the low-energy conformers based on their ab initio energies were used to produce weighting factors for the individual calculated shielding values to calculate the weighted average of the shielding values for a complete set of conformers. The differences in ¹H chemical shifts between the hydrogens of the two methyl groups and between the axial and equatorial hydrogens in 2-methylene-8,8-dimethyl-1,4,6,10-tetraoxaspiro[4.5] decane were shown to be due to energy differences between the chair and boat orientations of the six-membered ring and contribution from a twist-boat conformation. Results suggest a hypothesis that intramolecular differences in chemical shift might be calculated to a greater degree of accuracy than chemical shifts calculated relative to a standard.     

Oil Spill Dispersant Effectiveness Protocol. I: Impact of Operational Variables

The current U.S. Environmental Protection Agency protocol for testing the effectiveness of dispersants, the swirling flask test, has been found to give widely varying results in the hands of different testing laboratories. The sources of the ambiguities in the test were determined by considering several options in a rigorous systematic fashion. Options considered were variability among three operators and the variability and limitations of three analytical instruments in establishing a calibration curve for oil/dispersant mixtures. Other options included flask type, impact of operational variables (rotational speed, mixing time, settling time, and oil:dispersant ratio), development of a revised procedure for dispersant effectiveness, reproducibility and repeatability of the revised procedure, and development of selection criteria for screening of dispersants. A redesign of the test flask, which is characterized by having baffles with a stopcock at the bottom for sample collection, was deemed necessary to accomplish reproducibility within operator and between operators. The results indicated that the baffled flask provided a total coefficient of variation (variations within operator and between operators) less than 11% compared to above 100% in some cases for the swirling flask.     

Oil Spill Dispersant Effectiveness Protocol. II: Performance of Revised Protocol

The current U.S. Environmental Protection Agency (EPA) protocol for testing the effectiveness of dispersants for use in treating oil spills on the open water, the swirling flask test (SFT), has been found to give widely varying results in the hands of different testing laboratories. Part I addressed the sources of the ambiguities in the EPA SFT and the development of a new test referred to as the baffled flask test (BFT). In this part of the series, further experiments were conducted for estimating the repeatability of three operators in determining the effectiveness of 18 dispersants by both the EPA SFT and the BFT methods. Overall statistical analysis of the results indicated that the coefficient of variation by the BFT was only 7.8% compared to 21.9% for the EPA SFT. The mean percent effectiveness of the EPA SFT was only 19.7% as compared to 64.6% for the BFT. Proposed selection criteria for screening of dispersants for listing on the National Contingency Plan Product Schedule have been developed.     

Synthesis and characterization of poly(methylphenylsilylene)–poly(ethylene oxide) and poly(methylphenylsilylene)–polyisoprene multiblock copolymers

Multiblock copolymers were synthesized through condensation reactions of end‐groups of α,ω‐dichloro‐poly(methylphenylsilylene) with hydroxyl end‐groups of poly(ethylene glycol) or the chain‐ends of ‘living’ polyisoprenyl disodium. Optimum conditions have been sought through kinetic studies and by investigation of model reactions. The overall molecular weight distribution of poly(methylphenylsilylene)‐block‐poly(ethylene oxide) is characterized in terms of Flory's theory of condensation reactions, while the limiting step in the reaction is tentatively attributed to the formation of aggregates. © 2001 Society of Chemical Industry     

A new enzymatically produced 1-lactulose: A pilot study to test the bifidogenic effects

In a novel pilot plant scale process, lactulose was synthesized enzymatically, operating a maximum batch volume of 170 L. The test product was subjected to a randomized, double-blind, placebo-controlled pilot feeding study. Participants consumed a milk serum drink for 14 days, equalling a daily intake of about 3 g d⁻¹ galactosyl-fructoses. Additionally, the effect of industrially produced lactulose and a placebo drink without lactulose was determined. So far, clinical studies on the prebiotic action of lactulose only focussed on the effects of industrially generated lactulose. The intake of enzymatically synthesized lactulose significantly increased (p = 0.001) faecal bifidobacterial counts, while there was no difference with industrially produced lactulose. Analysis of the compounds showed that the drink with enzymatically synthesized lactulose contained an additional disaccharide, β-1,1-galactosyl-fructose. Presumably, this sugar has a bifidogenic effect and may represent a novel prebiotic.