Proteomic identification of salivary transferrin as a biomarker for early detection of oral cancer

Oral cancer has a low five-year survival rate. Early detection of oral cancer could reduce the mortality and morbidity associated with this disease. Saliva, which can be sampled non-invasively and is less complex than blood, is a good potential source of oral cancer biomarkers. Proteomic analysis of saliva from oral cancer patients and control subjects was performed to identify salivary biomarkers of early stage oral cancer in humans. The protein profile of pooled salivary samples from patients with oral squamous cell carcinoma (OSCC) or OSCC-free control subjects was analyzed using two-dimensional gel electrophoresis (2DE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analyses. Potential biomarkers were verified by Western blotting and ELISA assays. Transferrin levels were elevated in the saliva of OSCC patients as determined using 2DE followed by MALDI-TOF MS and confirmed by MALDI-TOF/TOF MS, Western blotting and ELISA. The increase in salivary transferrin levels in OSCC patients strongly correlated with the size and stage of the tumor. The area under the receiver-operating characteristics curves showed that salivary transferrin-based ELISA was highly specific, sensitive and accurate for the early detection of oral cancer. We have identified salivary transferrin as a biomarker for the detection of early stage oral cancer. This finding provides a promising basis for the development of a non-invasive diagnostic test for early stage oral cancer.     

Selective enrichment and identification of azide-tagged cross-linked peptides using chemical ligation and mass spectrometry

Protein-protein interaction is one of the key regulatory mechanisms for controlling protein function in various cellular processes. Chemical cross-linking coupled with mass spectrometry has proven to be a powerful method not only for mapping protein-protein interactions of all natures, including weak and transient ones, but also for determining their interaction interfaces. One critical challenge remaining in this approach is how to effectively isolate and identify cross-linked products from a complex peptide mixture. In this work, we have developed a novel strategy using conjugation chemistry for selective enrichment of cross-linked products. An azide-tagged cross-linker along with two biotinylated conjugation reagents were designed and synthesized. Cross-linking of model peptides and cytochrome c as well as enrichment of the resulting cross-linked peptides has been assessed. Selective conjugation of azide-tagged cross-linked peptides has been demonstrated using two strategies: copper catalyzed cycloaddition and Staudinger ligation. While both methods are effective, Staudinger ligation is better suited for enriching the cross-linked peptides since there are fewer issues with sample handling. LC MSⁿ analysis coupled with database searching using the Protein Prospector software package allowed identification of 58 cytochrome c cross-linked peptides after enrichment and affinity purification. The new enrichment strategy developed in this work provides useful tools for facilitating identification of cross-linked peptides in a peptide mixture by MS, thus presenting a step forward in future studies of protein-protein interactions of protein complexes by cross-linking and mass spectrometry.     

Hydrogen‐Bonding Induced Cooperative Effect on the Energy Transfer in Helical Polynorbornenes Appended with Porphyrin‐Containing Amidic Alanine Linkers

Polynorbornenes appended with porphyrins containing a range of different linkers are synthesized. The use of bisamidic chiral alanine linkers between the pending porphyrins and the polymeric backbone has been shown to bring the adjacent porphyrin chromophores to more suitable orientation for exciton coupling owing to hydrogen bonding between the adjacent linkers. The hydrogen bonding between the adjacent pendants in these polymers may induce a cooperative effect and therefore render single‐handed helical structures for these polymers. Such a cooperative effect is reflected in the enhancement of FRET efficiencies between zinc–porphyrin and free base porphyrin in random copolymers.     

Analysis of amino acids and biogenic amines in breast cancer cells by capillary electrophoresis using polymer solutions containing sodium dodecyl sulfate

We describe simultaneous analysis of naphthalene-2,3-dicarboxaldehyde (NDA)-amino acid and NDA-biogenic amine derivatives by CE in conjunction with light-emitting diode-induced fluorescence detection using poly(ethylene oxide) (PEO) solutions containing sodium dodecyl sulfate (SDS). After sample injection, via EOF 0.1% PEO prepared in 100 mM TB solution (pH 9.0) containing 30 mM SDS entered a capillary filled with 0.5 M TB solution (pH 10.2) containing 40 mM SDS. Under this condition, 14 NDA-amino acid and NDA-amine derivatives were separated within 16 min, with high efficiency ((1.0–3.2) × 10⁵ theoretical plates) and sensitivity (LODs at S/N = 3 ranging from 2.06 to 19.17 nM). In the presence of SDS and PEO, analytes adsorption on the capillary wall was suppressed, leading to high efficiency and reproducibility. The intraday analysis RSD values (n = 3) of the mobilities for the analytes are less than 0.52%. We have validated the practicality of this approach by quantitative determination of 9 amino acids in breast cancer cells (MCF-7) and 10 amino acids in normal epithelial cells (H184B5F5/M10). The concentrations of Tau and Gln in the MCF-7 cells were different than those in the H184B5F5/M10 cells, respectively. Our results show the potential of this approach for cancer study.     

Coherent control of nanoscale light localization in metamaterial: creating and positioning isolated subwavelength energy hot spots

We show the strong optically induced interactions between discrete metamolecules in a metamaterial system and coherent monochromatic continuous light beam with a spatially tailored phase profile can be used to prepare a subwavelength scale energy localization. Well-isolated energy hot spots of a fraction of a wavelength can be created and positioned on the metamaterial landscape offering new opportunities for data storage and imaging applications.     

Octaarginine-modified chitosan as a nonviral gene delivery vector: properties and in vitro transfection efficiency

Protein transduction domains (PTD) have been identified to have the capacity to facilitate molecular cargo to translocate through cell membrane. This study aims to utilize the cell membrane penetrating ability of octaarginine oligopeptide, a simplified prototype of the PTD, to enhance the transfection efficiency of chitosan. Octaarginine-modified chitosan (R₈-CS) was synthesized as a gene transfer carrier by carbodiimide chemistry. The structure and composition of R₈-CSs were characterized using FTIR and ¹H NMR. Agarose gel electrophoresis assay showed that R₈-CS could efficiently condense the DNA. The particle size of R₈-CS/DNA complexes were determined to be around 100–200 nm. The nanoparticle complexes exhibited a spherical and compact morphology. R₈-CS demonstrated higher transfection activity and lower cytotoxicity as compared to the unmodified chitosan and also showed good serum resistance.     

The temperature evolution of the magnetic correlations in pure and diluted spin ice Ho2−xYxTi2O7

Diffuse polarized neutron scattering studies have been carried out on single crystals of pyrochlore spin ice Ho_2−xY_xTi₂O₇ (x=0, 0.3, and 1) to investigate the effects of doping and anisotropy on spin correlations in the system. The crystals were aligned with the (1 −1 0) orientation coincident with the direction of neutron polarization. For all the samples studied the spin flip (SF) diffuse scattering (i.e. the in-plane component) reveals that the spin correlations can be described using a nearest-neighbour spin ice model (NNSM) at higher temperatures (T=3.6 K) and a dipolar spin ice model (DSM) as the temperature is reduced (T=30 mK). In the non-spin flip (NSF) channel (i.e. the out-of-plane component), the signature of strong antiferromagnetic correlations is observed for all the samples at the same temperature as the dipolar spin ice behaviour appears in the SF channel. Our studies show that the non-magnetic dopant Y does not significantly alter SF or NSF scattering for the spin ice state, even when Y doping is as high as 50%. In this paper, we focus on the experimental results of the highly doped spin ice HoYTi₂O₇ and compare our results with pure spin ice Ho₂Ti₂O₇. The crossover from a dipolar to a nearest-neighbour spin ice behaviour and the doping insensitivity in spin ices are briefly discussed.     

Unruh effect and holography

We study the Unruh effect on the dynamics of quarks and mesons in the context of AdS₅/CFT₄ correspondence. We adopt an AdS₅ metric with the boundary Rindler horizon extending into a bulk Rindler-like horizon, which yields the thermodynamics with Unruh temperature verified by computing the boundary stress tensor. We then embed in it a probe fundamental string and a D7 brane in such a way that they become the dual of an accelerated quark and a meson in Minkowski space, respectively. Using the standard procedure of holographic renormalization, we calculate the chiral condensate, and also the spectral functions for both the accelerated quark and meson. Especially, we extract the corresponding strength of random force of the Langevin dynamics and observe that it can characterize the phase transition of meson melting. This result raises an issue toward a formulation of complementarity principle for the Rindler horizon. We find most of the dynamical features are qualitatively similar to the ones in the thermal bath dual to the AdS black hole background, though they could be quite different quantitatively.     

On the dynamics of radially symmetric granulomas

A granuloma is a collection of macrophages that contains bacteria or other foreign substances that the body?s immune response is unable to eliminate. In this paper we present a simple mathematical model of radially symmetric granuloma dynamics. The model consists of a coupled system of two semi-linear parabolic equations for the macrophage density, and the bacterial density. The boundary of the granuloma is free. This simple framework makes it possible to conduct a mathematical analysis of the system dynamics. In particular, we show that the model system has a unique solution, and that, depending on the biological parameters; the bacterial load either disappears over time or persists. We use numerical methods to establish the existence of stationary solutions and examine how a stationary solution changes with the reproductive rate of the bacteria. These simulations show that the structure of the granuloma breaks down as the reproductive rate of the bacteria increases.