Exploring the interaction of ruthenium(II) polypyridyl complexes with DNA using single-molecule techniques

Here we explore DNA binding by a family of ruthenium(II) polypyridyl complexes using an atomic force microscope (AFM) and optical tweezers. We demonstrate using AFM that Ru(bpy)2dppz2+ intercalates into DNA (K(b) = 1.5 x 10(5) M(-1)), as does its close relative Ru(bpy)2dppx2+ (K(b) = 1.5 x 10(5) M(-1)). However, intercalation by Ru(phen)3(2+) and other Ru(II) complexes with K(b) values lower than that of Ru(bpy)2dppz2+ is difficult to determine using AFM because of competing aggregation and surface-binding phenomena. At the high Ru(II) concentrations required to evaluate intercalation, most of the DNA strands acquire a twisted, curled conformation that is impossible to measure accurately. The condensation of DNA on mica in the presence of polycations is well known, but it clearly precludes the accurate assessment by AFM of DNA intercalation by most Ru(II) complexes, though not by ethidium bromide and other monovalent intercalators. When stretching individual DNA molecules using optical tweezers, the same limitation on high metal concentration does not exist. Using optical tweezers, we show that Ru(phen)2dppz2+ intercalates avidly (K(b) = 3.2 x 10(6) M(-1)) whereas Ru(bpy)3(2+) does not intercalate, even at micromolar ruthenium concentrations. Ru(phen)3(2+) is shown to intercalate weakly (i.e., at micromolar concentrations (K(b) = 8.8 x 10(3) M(-1))). The distinct differences in DNA stretching behavior between Ru(phen)3(2+) and Ru(bpy)3(2+) clearly illustrate that intercalation can be distinguished from groove binding by pulling the DNA with optical tweezers. Our results demonstrate both the benefits and challenges of two single-molecule methods of exploring DNA binding and help to elucidate the mode of binding of Ru(phen)3(2+).     

Assessment of photosensitizer dosimetry and tissue damage assay for photodynamic therapy in advanced-stage tumors

Photodynamic therapy (PDT) efficacy is a complex function of tissue sensitivity, photosensitizer (PS) uptake, tissue oxygen concentration, delivered light dose and some other parameters. To better understand the mechanisms and optimization of PDT treatment, we assessed two techniques for quantifying tissue PS concentration and two methods for quantifying pathological tumor damage. The two methods used to determine tissue PS concentration kinetic were in vivo fluorescence probe and ex vivo chemical extraction. Both methods show that the highest tumor to normal tissue PS uptake ratio appears 4 h after PS administration. Two different histopathologic techniques were used to quantify tumor and normal tissue damage. A planimetry assessment of regional tumor necrosis demonstrated a linear relationship with increasing light dose. However, in large murine tumors this finding was complicated by the presence of significant spontaneous necrosis. A second method (densitometry) assessed cell death by nuclear size and density. With some exceptions the densitometry method generally supported the planimetry results. Although the densitometry method is potentially more accurate, it has greater potential subjectivity. Finally, our research suggests that the tools or methods we are studying for quantifying PS levels and tissue damage are necessary for the understanding of PDT effect and therapeutic ratio in experimental in vivo tumor research.     

Negative ion laser desorption ionization time-of-flight mass spectrometry of nitrated polycyclic aromatic hydrocarbons

Nitrated polycyclic aromatic hydrocarbons (nitro-PAHs) are widespread environmental pollutants that are generated by incomplete combustion and by atmospheric transformation of polycyclic aromatic hydrocarbons (PAHs). Many nitro-PAH compounds are potent genotoxins and some are direct acting mutagens. Detection of nitro-PAHs in aerosols is complicated by small sample sizes and nitro-PAH abundances that are 1–2 orders of magnitude less than analogous unsubstituted PAHs. Selective detection of several nitro-PAHs by using laser desorption ionization time-of-flight mass spectrometry in negative ion mode has been achieved. Desorption and ionization of nitro-PAHs were effected by using pulsed UV radiation at 266 and 213 ran. Intense molecular anions were observed in addition to fragments identified as CN^? and NO ₂ ^? , which were characteristic indicators of the presence of nitro-PAHs. Selective detection of nitro-PAHs in negative ion mode was demonstrated in the analysis of a diesel particulate sample.     

Airborne Nanostructured Particles and Occupational Health

Nanotechnology is leading to the development in many field, of new materials and devices in many fields that demonstrate nanostructure-dependent properties. However, concern has been expressed that these same properties may present unique challenges to addressing potential health impact. Airborne particles associated with engineered nanomaterials are of particular concern, as they can readily enter the body through inhalation. Research into the potential occupational health risks associated with inhaling engineered nanostructured particles is just beginning. However, there is a large body of data on occupational and environmental aerosols, which is applicable to developing an initial assessment of potential risk and risk reduction strategies. Epidemiological and pathological studies of occupational and environmental exposures to airborne particles and fibers provide information on the aerosol-related lung diseases and conditions that have been observed in humans. Toxicological studies provide information on the specific disease mechanisms, dose–response relationships, and the particle characteristics that influence toxicity, including the size, surface area, chemistry or reactivity, solubility, and shape. Potential health risk will depend on the magnitude and nature of exposures to airborne nanostructured particles, and on the release, dispersion, transformation and control of materials in the workplace. Aerosol control methods have not been well-characterized for nanometer diameter particles, although theory and limited experimental data indicate that conventional ventilation, engineering control and filtration approaches should be applicable in many situations. Current information supports the development of preliminary guiding principles on working with engineered nanomaterials. However critical research questions remain to be answered before the potential health risk of airborne nanostructured particles in the workplace can be fully addressed. Disclaimer The findings and conclusions in this report are those of the authors and do not necessarily represent the views of the National Institute for Occupational Safety and Health     

Copper-mediated aerobic synthesis of 3-azabicyclo[3.1.0]hex-2-enes and 4-carbonylpyrroles from N-allyl/propargyl enamine carboxylates

Synthetic methods for 3-azabicyclo[3.1.0]hex-2-enes and 4-carbonylpyrroles have been developed that use copper-mediated/catalyzed reactions of N-allyl/propargyl enamine carboxylates under an O(2) atmosphere and involve intramolecular cyclopropanation and carbooxygenation, respectively. These methodologies take advantage of orthogonal modes of chemical reactivity of readily available N-allyl/propargyl enamine carboxylates; the complementary pathways can be accessed by slight modification of the reaction conditions.     

Reduced decompositions and permutation patterns

Billey, Jockusch, and Stanley characterized 321-avoiding permutations by a property of their reduced decompositions. This paper generalizes that result with a detailed study of permutations via their reduced decompositions and the notion of pattern containment. These techniques are used to prove a new characterization of vexillary permutations in terms of their principal dual order ideals in a particular poset. Additionally, the combined frameworks yield several new results about the commutation classes of a permutation. In particular, these describe structural aspects of the corresponding graph of the classes and the zonotopal tilings of a polygon defined by Elnitsky that is associated with the permutation.     

Preservice Elementary Teachers' Attitudes Toward Their Past Experience With Science Fairs

The purpose of this research was to explore the experiences preservice elementary teachers have had with science fairs, to examine the attitudes resulting from their participation in science fairs as students, and to assess the importance of these past histories for their future students. Preservice teachers were asked what they liked, disliked, and what they considered to be the benefits of science fairs. While most felt that science fairs are beneficial activities for students, those who rated science fairs as most worthwhile had never participated in a science fair as either an elementary or a secondary student. Based on the results of the study, several recommendation were made to improve science fair experiences both for teachers and for students.     

Integrated paramagnetic resonance of high-spin Co(II) in axial symmetry: chemical separation of dipolar and contact electron-nuclear couplings

Integrated paramagnetic resonance, utilizing electron paramagnetic resonance (EPR), NMR, and electron-nuclear double resonance (ENDOR), of a series of cobalt bis-trispyrazolylborates, Co(Tp ( x )) 2, are reported. Systematic substitutions at the ring carbons and on the apical boron provide a unique opportunity to separate through-bond and through-space contributions to the NMR hyperfine shifts for the parent, unsubstituted Tp complex. A simple relationship between the chemical shift difference (delta H - delta Me) and the contact shift of the proton in that position is developed. This approach allows independent extraction of the isotropic hyperfine coupling, A iso, for each proton in the molecule. The Co..H contact coupling energies derived from the NMR, together with the known metrics of the compounds, were used to predict the ENDOR couplings at g perpendicular. Proton ENDOR data is presented that shows good agreement with the NMR-derived model. ENDOR signals from all other magnetic nuclei in the complex ( (14)N, coordinating and noncoordinating, (11)B and (13)C) are also reported.