Quantification of 5-methylcytosine, 5-hydroxymethylcytosine and 5-carboxylcytosine from the blood of cancer patients by an enzyme-based immunoassay

Background

Genome-wide aberrations of the classic epigenetic modification 5-methylcytosine (5mC), considered the hallmark of gene silencing, has been implicated to play a pivotal role in mediating carcinogenic transformation of healthy cells. Recently, three epigenetic marks derived from enzymatic oxidization of 5mC namely 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC), have been discovered in the mammalian genome. Growing evidence suggests that these novel bases possess unique regulatory functions and may play critical roles in carcinogenesis.

Methods

To provide a quantitative basis for these rare epigenetic marks, we have designed a biotin–avidin mediated enzyme-based immunoassay (EIA) and evaluated its performance in genomic DNA isolated from blood of patients diagnosed with metastatic forms of lung, pancreatic and bladder cancer, as well as healthy controls. The proposed EIA incorporates spatially optimized biotinylated antibody and a high degree of horseradish-peroxidase (HRP) labeled streptavidin, facilitating signal amplification and sensitive detection.

Results

We report that the percentages of 5mC, 5hmC and 5caC present in the genomic DNA of blood in healthy controls as 1.025 ± 0.081, 0.023 ± 0.006 and 0.001 ± 0.0002, respectively. We observed a significant (p < 0.05) decrease in the mean global percentage of 5hmC in blood of patients with malignant lung cancer (0.013 ± 0.003%) in comparison to healthy controls.

Conclusion

The precise biological roles of these epigenetic modifications in cancers are still unknown but in the past two years it has become evident that the global 5hmC content is drastically reduced in a variety of cancers. To the best of our knowledge, this is the first report of decreased 5hmC content in the blood of metastatic lung cancer patients and the clinical utility of this observation needs to be further validated in larger sample datasets.     

Multichromophoric Organic Molecules Encapsulated in Polymer Nanoparticles for Artificial Light Harvesting

We designed a self‐assembled multichromophoric organic molecular arrangement inside polymer nanoparticles for light‐harvesting antenna materials. The self‐assembled molecular arrangement of quaterthiophene molecules was found to be an efficient light‐absorbing antenna material, followed by energy transfer to Nile red (NR) dye molecules, which was confined in polymer nanoparticles. The efficiency of the antenna effect was found to be 3.2 and the effective molar extinction coefficient of acceptor dye molecules was found to be enhanced, which indicates an efficient light‐harvesting system. Based on this energy‐transfer process, tunable photo emission and white light emission has been generated with 14 % quantum yield. Such self‐assembled oligothiophene–NR systems encapsulated in polymer nanoparticles may open up new possibilities for fabrication of artificial light harvesting system.     

Self‐assembly of silver nanocolloids in the Langmuir–Blodgett Film of stearic acid: Evidence of an efficient SERS sensing platform

Highly sensitive surface‐enhanced Raman scattering active substrate obtained by self‐assembly of silver nanocolloids (AgNCs) in the bilayer Langmuir–Blodgett (LB) film of stearic acid (SA) has been reported. Rhodamine 6G (R6G) has been used as the probe molecule to test the efficacy of the as prepared substrate. Gigantic enhancement factors ~10¹² orders of magnitude have been estimated from the surface‐enhanced resonance Raman scattering [SER(R) S] spectrum of R6G, which proves that the as prepared substrate is superior or comparable with silver nanoparticle as dried AgNC solutions on microscopic slides. The optical properties of the as prepared substrates have been envisaged by ultraviolet‐visible absorption spectra, while their morphological features are mapped through field emission scanning electron microscopy (FESEM) and atomic force microscopy (AFM) images. A correlation has been drawn between the SER(R) S efficacy and the corresponding FESEM and AFM images of the as prepared substrates. Electric field distributions around the aggregated AgNCs have been estimated with the aid of three‐dimensional finite difference time domain simulation studies. Localized surface plasmon coupling between the nanoaggregated geometries may be altered by lifting the LB film of SA at various surface pressures. Copyright © 2015 John Wiley & Sons, Ltd.     

Singular solutions and Green's method in micropolar theory of elasticity

The matrices of fundamental solutions are constructed for a concentrated force as well as a concentrated couple varying harmonically in time and acting in an unbounded micropolar elastic continuum. These solutions are then used to obtain solutions for some other loading singularities. Integral representations, for the displacement and the rotation vectors are obtained by making use of the basic singular solutions. The formal solutions to two fundamental boundary value problems are expressed in terms of integrals which include given surface and body data and Green's functions.     

Thermal stresses due to uniform temperature distributed over a band of the cylindrical hole in an infinite body

Summary Two-dimensional thermal stress problems involving plates, spheres and specially circular cylinders have received the bulk of the attention in the literature by various authors. In this paper the thermal stresses in an infinite isotropic elastic solid have been obtained when a band of the internal surface of a cylindrical hole in the solid is kept at a constant temperature.     

Studies on solution and solution-cast film of polyaniline colloids prepared in the absence and presence of ultrasonic irradiation

Spectroscopic and voltammetric features for the solution and solution-cast film of polyaniline (PANI) colloids prepared in the absence and presence of ultrasonic irradiation were examined. Significant differences in the color and absorption spectra between the solution of colloids prepared in the absence and presence of irradiation were observed. Effects of ambient atmosphere on the solution of colloids were also studied. A gradual decay in the absorption at approximately 365 nm was occurred upon ageing the solution in air for the colloid prepared under ultrasonic irradiation. PANI film, cast from its colloidal solution, exhibited excellent electroactivity. The films thus cast were applied to electrode modification to control electrochemical reaction. Clear voltammetric response was observed on the electrode modified with the colloid prepared in the presence of ultrasonic irradiation.     

SH-type waves dispersion in an isotropic medium sandwiched between an initially stressed orthotropic and heterogeneous semi-infinite media

The present paper studies the propagation of shear waves (SH-type waves) in an homogeneous isotropic medium sandwiched between two semi infinite media. The upper half-space is considered as orthotropic medium under initial stress and lower half-space considered as heterogeneous medium. We have obtained the dispersion equation of phase velocity for SH-type waves. The propagation of SH-type waves are influenced by inhomogeneity parameters and initial stress parameter. The velocity of SH-type wave has been computed for different cases. We have also obtained the dispersion equation of phase velocity in homogeneous media in the absence of initial stress. The velocities of SH-type waves are calculated numerically as a function of kH (non-dimensional wave number) and presented in a number of graphs. To study the effect of inhomogeneity parameters and initial stress parameter we have plotted the velocity of SH-type wave in several figure. We have observed that the velocity of wave increases with the increase inhomogeneity parameters. We found that in both homogeneous and inhomogeneous media the velocity of SH-type wave increases with the increase of initial stress parameter. The results may be useful for the study of seismic waves propagation during any earthquake and artificial explosions.     

Access to Different Nanostructures via Self‐Assembly of Thiourea‐Containing PEGylated Amphiphiles

Readily water‐soluble PEGylated amphiphiles containing bis‐thiourea‐based molecular recognition units at the interface of hydrophobic and hydrophilic blocks are developed. Self‐assembly of these amphiphiles is found to be dependent on the exact chemical composition of the hydrophobic component. Elongated, spherical, and disk‐like micelles are formed with the change in hydrophobic group from stearyl (2A), oleyl (2B), and dodecanol (2C), respectively. The length of the rod‐like elongated micelles formed by 2A could be tuned by thermal treatment as well. Synthesis and detailed structural characterization of these amphiphiles by TEM, DSC, synchrotron SAXS techniques are reported. Organic solvent‐free direct aqueous encapsulation of doxorubicin, an anticancer drug into these nanostructures is demonstrated.

     

The effect of inertia on the Dirac electron,the spin Hall current and the momentum space Berry curvature

We have studied the spin dependent force and the associated momentum space Berry curvature in an accelerating system. The results are derived by taking into consideration the non-relativistic limit of a generally covariant Dirac equation with an electromagnetic field present, where the methodology of the Foldy–Wouthuysen transformation is applied to achieve the non-relativistic limit. Spin currents appear due to the combined action of the external electric field, the crystal field and the induced inertial electric field via the total effective spin–orbit interaction. In an accelerating frame, the crucial role of momentum space Berry curvature in the spin dynamics has also been addressed from the perspective of spin Hall conductivity. For time dependent acceleration, the expression for the spin polarization has been derived.