New bioluminogenic substrates for monoamine oxidase assays

Novel bioluminogenic substrates were designed for probing monoamine oxidase (MAO) activity based on a simple and effective beta-elimination strategy. By modifying the amino group and the central core of luciferin derivatives, we have developed a series of substrates useful for assays of MAO A or B, or both. One of these substrates, exhibiting low Km values and high signal-to-background ratios with both isozymes, was shown to accurately measure the Ki values of known MAO inhibitors. This substrate is a key component in the development of a highly sensitive homogeneous MAO assay for high-throughput screening (HTS) of compounds in drug discovery and for monitoring MAO activity in complex biological systems. This design strategy should be applicable to fluorogenic MAO substrates and could broaden the structural requirements of substrates for other enzyme assays.     

Schubert polynomials and classes of Hessenberg varieties

Regular semisimple Hessenberg varieties are a family of subvarieties of the flag variety that arise in number theory, numerical analysis, representation theory, algebraic geometry, and combinatorics. We give a “Giambelli formula” expressing the classes of regular semisimple Hessenberg varieties in terms of Chern classes. In fact, we show that the cohomology class of each regular semisimple Hessenberg variety is the specialization of a certain double Schubert polynomial, giving a natural geometric interpretation to such specializations. We also decompose such classes in terms of the Schubert basis for the cohomology ring of the flag variety. The coefficients obtained are nonnegative, and we give closed combinatorial formulas for the coefficients in many cases. We introduce a closely related family of schemes called regular nilpotent Hessenberg schemes, and use our results to determine when such schemes are reduced.     

Impact of Photodynamic Treatment with Meso-substituted Porphyrin on the Immunomodulatory Capacity of White Blood Cell-containing Red Blood Cell Products

After transfusion, the presence of contaminating white blood cells (WBC) in blood components may result in either deleterious or positive immunological responses. We have previously reported that photodynamic treatment (PDT) with meso-substituted mono-phenyl-tri-( N -methyl-4-pyridyl)-porphyrin (Tri-P(4)) and red light can inactivate pathogens in red blood cell (RBC) products. The present study explored the effect of PDT on contaminating WBC in RBC products with varying hematocrit (Hct). After PDT, we evaluated adaptive and innate immunomodulation through allogeneic and mitogenic stimulation. PDT resulted in decreased T-cell proliferation which was more pronounced with lower Hct. Dark effect of porphyrin Tri-P(4) was remarkable on antigen-presenting cells affecting expression of co-stimulatory molecules CD80/CD86. Finally, cytokine profile after PDT revealed a mixed Th1/Th2 type response while surface antigen expression supported the development of alternatively activated macrophages (AAMφ or Type 2 macrophages) instead of dendritic cells. In conclusion, PDT with Tri-P(4) altered proliferation, allo-stimulation, cell surface antigen expression and cytokine profiles of the cells. These results suggest that PDT may be potentially useful in preventing transfusion-associated graft- versus -host disease and alloimmunization. It seems worthwhile to further explore PDT-induced immunomodulation to optimize conditions which may result in allo-tolerance by AAMφ.     

Gravitational and Electroweak Unification

Einstein suggested that a unified field theorybe constructed by replacing the diffeomorphisms (thecoordinate transformations of general relativity) withsome larger group. We have constructed a theory that unifies the gravitational and electroweakfields by replacing the diffeomorphisms with the largestgroup of coordinate transformations under whichconservation laws are covariant statements. Thisreplacement leads to a theory with field equations whichimply the validity of the Einstein equations of generalrelativity, with a stress-energy tensor that is justwhat one expects for the electroweak field andassociated currents. The electroweak field appears as aconsequence of the field equations (rather than as a"compensating field" introduced to secure gaugeinvariance). There is no need for symmetry breaking toaccommodate mass, because the U(1) × SU(2) gaugesymmetry is approximate from the outset. Thegravitational field is described by the space-timemetric, as in general relativity. The electroweak fieldis described by the "mixed symmetry" part of the Riccirotation coefficients. The gauge symmetry-breakingquantity is a vector formed by contracting theLevi-Civita symbol with the totally antisymmetric partof the Ricci rotation coefficients.     

Reversed-phase HPLC of polymer additives with multiple on-line spectroscopic analysis (UV,IR, (1)H NMR and MS)

The reversed-phase chromatography of a number of polymer additives has been undertaken with on-line characterisation via a combination of diode array UV, (1)H NMR, IR spectroscopy and mass spectrometry. This combination of spectrometers enabled the on-flow collection of full UV, (1)H NMR, IR and mass spectra for a range of common polymer additives in amounts ranging from ca. 230 to 900 micro g on-column. The practical difficulties associated with multiple hyphenation and potential future developments are discussed.     

Synthesis and characterization of 5-substituted 1,3-diazacyclohexane derivatives

Three synthetic routes to 5-substituted 1,3-diazacyclohexane derivatives 1 are reported. The first method involves treatment of 1, 3-diaminopropan-2-ol 2 with paraformaldehyde to yield 5-hydroxy-1, 3-diazacyclohexane 3. A second method is based on the condensation of 2-bromo-2-nitro-1,3-propanediol with tert-butylamine and formaldehyde to yield 1,3-di-tert-butyl-5-bromo-5-nitro-1, 3-diazacyclohexane 22. The third method relies on the cycloalkylation of methylenebisacetamide with 3-chloro-2-chloromethyl-2-propene to provide 5-exomethylene-1, 3-diacetyl-1,3-diazacyclohexane 28. Functional group manipulations of 3, 22, and 28 provide a number of novel 1,3-diazacyclohexanes functionalized at the 5-position.     

Reaction of nitriles under acidic conditions. Part I. A general method of synthesis of condensed pyrimidines

Nitriles are known to give rise to salts of different compositions with halogen acids. Many of the reactions undergone by nitriles under the influence of halogen acids are, in many cases, assumed to proceed via the intermediate formation of highly reactive imidoyl derivatives. The intermediates produced, in situ, by the reaction of nitriles with hydrogen chloride should, in principle, be capable of reacting with compounds containing appropriately placed nucleophilic and electrophilic centers leading to the formation of a heterocycle incorporating the C=N of the nitrile. Thus, the reaction of aliphatic, aromatic and heterocyclic nitriles with a variety of ortho aminocarbonyl derivatives such as nitriles, amides, esters and ketones of benzene, thiophen, furan, pyrrole, benzothiophene and pyridothiophene have been found to yield the corresponding condensed pyrimidines in fair to good yields. This constitutes a facile and versatile one-pot synthesis of condensed pyrimidines.