Mark Lowry

Selective fluorescence detection of cysteine and N-terminal cysteine peptide residues.

A new fluorogenic fluorescein derivative containing an alpha,beta-unsaturated aldehyde moiety produces a selective fluorescent signal enhancement in the presence of cysteine or peptides containing N-terminal cysteine residues. The mechanism is based on synergistic covalent and supramolecular interactions.

Seminaphthofluorones are a family of water-soluble, low molecular weight, NIR-emitting fluorophores

A readily accessible new class of near infrared (NIR) molecular probes has been synthesized and evaluated. Specific fluorophores in this unique xanthene based regioisomeric seminaphthofluorone dye series exhibit a combination of desirable characteristics including (i) low molecular weight (339 amu), (ii) aqueous solubility, and (iii) dual excitation and emission from their fluorescent neutral and anionic forms. Importantly, systematic changes in the regiochemistry of benzannulation and the ionizable moieties afford (iv) tunable deep-red to NIR emission from anionic species and (v) enhanced Stokes shifts. Anionic SNAFR-6, exhibiting an unusually large Stokes shift of ≈200 nm (5,014 cm−1) in aqueous buffer, embodies an unprecedented fluorophore that emits NIR fluorescence when excited in the blue/green wavelength region. The successful use of SNAFR-6 in cellular imaging studies demonstrates proof-of-concept that this class of dyes possesses photophysical characteristics that allow their use in practical applications. Notably, each of the new fluorophores described is a minimal template structure for evaluation of their basic spectral properties, which may be further functionalized and optimized yielding concomitant improvements in their photophysical properties.

A Fast Response Highly Selective Probe for the Detection of Glutathione in Human Blood Plasma

A fluorescent probe for glutathione (GSH) detection was developed. Our study indicates a possible mechanism which couples a conjugate addition and micelle-catalyzed large membered ring formation/elimination sequence. This method enables excellent selectivity towards GSH over other biological thiols such as cysteine (Cys) and homocysteine (Hcy). The proposed method is precise with a relative standard deviation (R.S.D) lower than 6% (n = 3) and has been successfully applied to determine GSH in human plasma with recoveries between 99.2% and 102.3%.

Controllable formation of ionic liquid micro- and nanoparticles via a melt-emulsion-quench approach.

We present a facile, scalable, and general method for the size-variable generation of monodispersed, near-spherical solid-state (frozen) ionic liquid nanoparticles based on a novel melt-emulsion-quench approach. Simple manipulation of the internal templating droplets within oil-in-water (o/w) microemulsions also permits the formation of well-defined microspheres. This simple and rapid preparation, requiring neither specialized equipment nor harsh conditions, suggests a wealth of potential for these designer nanomaterials within the biomedical, materials, and analytical communities.

Gold Nanoparticle Sensor for Homocysteine Thiolactone-Induced Protein Modification

Homocysteine thiolactone-induced protein modification (HTPM) is a unique post-translational protein modification that is recognized as an emergent biomarker for cardiovascular disease. HTPM involves the site-specific acylation of proteins at lysine residues by homocysteine thiolactone (HTL) to produce protein homocystamide, which has been found at elevated levels in patients with coronary heart disease. Herein, we report the development of a novel gold nanoparticle (GNP) biochemical sensor for detection of protein homocystamide in an in vitro serum protein-based model system. Human serum albumin (HSA) and human sera were subjected to HTPM in vitro to produce HSA-homocystamide or serum protein homocystamide, respectively, which was subsequently treated with citrate-capped GNPs. This GNP sensor typically provided instantaneous visual confirmation of HTPM in the protein model systems. Transmission electron microscopy images of the GNPs in the presence of HSA-homocystamide suggest that modification-directed nanoparticle assembly is the mechanism by which the biochemical sensor produces a colorimetric signal. The resultant nanoparticle-protein assembly exhibited excellent thermal and dilutional stability, which is expected for a system stabilized by chemisorption and intermolecular disulfide bonding. The sensor typically provided a linear response for modified human sera concentrations greater than approximately 5 mg/mL. The calculated limit of detection and calibration sensitivity for the method in human sera were 5.2 mg/mL and 13.6 AU . (microg/mL)-1, respectively.

Amino Acid-Based Fluorescent Chiral Ionic Liquid for Enantiomeric Recognition

We report on the synthesis and characterization of a new fluorescent chiral ionic liquid (FCIL), l-phenylalanine ethyl ester bis(trifluoromethane) sulfonimide (l-PheC(2)NTf(2)), capable of serving simultaneously as solvent, chiral selector, and fluorescent reporter in chiral analytical measurements. Enantiomers of different analytes, including fluorescent and nonfluorescent compounds, with a variety of structures were shown to induce wavelength- and analyte-dependent changes in the fluorescence intensity of this FCIL. This system may provide both chemo- and enantioselectivity toward multiple analytes simultaneously. The newly synthesized FCIL, derived from commercially available l-phenylalanine ethyl ester chloride and lithium bis(trifluoromethane) sulfonamide, was obtained as liquid at room temperature and is stable to thermal decomposition up to 270 degrees C. Absorption and fluorescence properties of neat l-PheC(2)NTf(2) were complex. While the absorption properties were similar to phenylalanine with a weakly absorbing tail extending beyond 400 nm, multiple excitation and emission bands were observed in its Excitation-Emission Matrix (EEM). A prominent excimer emission displayed the greatest intensity of all emission bands, and a long-wavelength emission shifted toward the red with increasing excitation wavelength. These different spectral regions were shown to respond differently toward several analytes, including sugars such as glucose and mannose, making this an ideal system to exploit the multidimensional properties of fluorescence. The unique properties of l-PheC(2)NTf(2) combined with EEMs resulted in reliable identification of different enantiomers and measurement of enantiomeric composition. Importantly, the choice of excitation and emission wavelength regions was an important variable shown to improve prediction of enantiomeric composition.

Rhein inhibits angiogenesis and the viability of hormone-dependent and -independent cancer cells under normoxic or hypoxic conditions in vitro

Hypoxia is a hallmark of solid tumors, including breast cancer, and the extent of tumor hypoxia is associated with treatment resistance and poor prognosis. Considering the limited treatment of hypoxic tumor cells and hence a poor prognosis of breast cancer, the investigation of natural products as potential chemopreventive anti-angiogenic agents is of paramount interest. Rhein (4,5-dihydroxyanthraquinone-2-carboxylic acid), the primary anthraquinone in the roots of Cassia alata L., is a naturally occurring quinone which exhibits a variety of biologic activities including anti-cancer activity. However, the effect of rhein on endothelial or cancer cells under hypoxic conditions has never been delineated. Therefore, the aim of this study was to investigate whether rhein inhibits angiogenesis and the viability of hormone-dependent (MCF-7) or -independent (MDA-MB-435s) breast cancer cells in vitro under normoxic or hypoxic conditions. Rhein inhibited vascular endothelial growth factor (VEGF(165))-stimulated human umbilical vein endothelial cell (HUVEC) tube formation, proliferation and migration under normoxic and hypoxic conditions. In addition, rhein inhibited in vitro angiogenesis by suppressing the activation of phosphatidylinositol 3-kinase (PI3K), phosphorylated-AKT (p-AKT) and phosphorylated extracellular signal-regulated kinase (p-ERK) but showed no inhibitory effects on total AKT or ERK. Rhein dose-dependently inhibited the viability of MCF-7 and MDA-MB-435s breast cancer cells under normoxic or hypoxic conditions, and inhibited cell cycle in both cell lines. Furthermore, Western blotting demonstrated that rhein inhibited heat shock protein 90alpha (Hsp90α) activity to induce degradation of Hsp90 client proteins including nuclear factor-kappa B (NF-κB), COX-2, and HER-2. Rhein also inhibited the expression of hypoxia-inducible factor-1 alpha (HIF-1α), vascular endothelial growth factor (VEGF(165)), epidermal growth factor (EGF), and the phosphorylation of inhibitor of NF-κB (I-κB) under normoxic or hypoxic conditions. Taken together, these data indicate that rhein is a promising anti-angiogenic compound for breast cancer cell viability and growth. Therefore, further studies including in vivo and pre-clinical need to be performed.

Differences in heterocycle basicity distinguish homocysteine from cysteine using aldehyde-bearing fluorophores

We report the detection of homocysteine over cysteine based upon characteristic differences between 5- and 6-membered heterocyclic amines formed upon reaction with aldehyde-bearing compounds. Homocysteine-derived thiazinane-4-carboxylic acids are more basic than cysteine-derived thiazolidines-4-carboxylic acids. Fluorescence enhancement in response to homocysteine is achieved by tuning pH and excitation wavelength.

Field Effects Induce Bathochromic Shifts in Xanthene Dyes

There is ongoing interest in near-infrared (NIR) absorbing and emitting dyes for a variety of biomedical and materials applications. Simple and efficient synthetic procedures enable the judicious tuning of through-space polar (field) effects as well as low barrier hydrogen bonding to modulate the HOMO-LUMO gap in xanthene dyes. This affords unique NIR-absorbing xanthene chromophores.

Fluorescence, Phosphorescence, and Chemiluminescence

Noureen Siraj,† Bilal El-Zahab,‡ Suzana Hamdan,† Tony E. Karam,† Louis H. Haber,† Min Li, Sayo O. Fakayode, Susmita Das, Bertha Valle, Robert M. Strongin, Gabor Patonay, Herman O. Sintim, Gary A. Baker, Aleeta Powe, Mark Lowry, Jan O. Karolin, Chris D. Geddes, and Isiah M. Warner*,† †Department of Chemistry, Louisiana State University, Baton Rouge, Louisiana 70803, United States ‡Department of Mechanical and Materials Engineering, Florida International University, Miami, Florida 33174, United States Process Development Center, Albemarle Corporation, Baton Rouge, Louisiana 70805, United States Department of Chemistry, Winston-Salem State University, Winston-Salem, North Carolina 27110, United States Department of Civil Engineering, Adamas Institute of Technology, Barasat, Kolkata 700126, West Bengal India Department of Chemistry, Texas Southern University, Houston, Texas 77004, United States Department of Chemistry, Portland State University, Portland, Oregon 97207, United States Department of Chemistry, Georgia State University, Atlanta, Georgia 30302-4098, United States Department of Chemistry and Biochemistry, University of Maryland, College Park, Maryland 20742, United States Department of Chemistry, University of Missouri Columbia, Columbia, Missouri 65211-7600, United States Department of Chemistry, University of Louisville, Louisville, Kentucky 40208, United States Institute of Fluorescence, University of Maryland Baltimore County, Baltimore, Maryland 21202, United States