Ranjana Mehrotra

Assessment of amsacrine binding with DNA using UV-visible, circular dichroism and Raman spectroscopic techniques

Proper understanding of the mechanism of binding of drugs to their targets in cell is a fundamental requirement to develop new drug therapy regimen. Amsacrine is a rationally designed anticancer drug, used to treat leukemia and lymphoma. Binding with cellular DNA is a crucial step in its mechanism of cytotoxicity. Despite numerous studies, DNA binding properties of amsacrine are poorly understood. Its reversible binding with DNA does not permit X-ray crystallography or NMR spectroscopic evaluation of amsacrine-DNA complexes. In the present work, interaction of amsacrine with calf thymus DNA is investigated at physiological conditions. UV-visible, FT-Raman and circular dichroism spectroscopic techniques were employed to determine the binding mode, binding constant, sequence specificity and conformational effects of amsacrine binding to native calf thymus DNA. Our results illustrate that amsacrine interacts with DNA by and large through intercalation between base pairs. Binding constant of the amsacrine-DNA complex was found to be K=1.2±0.1×10(4) M(-1) which is indicative of moderate type of binding of amsacrine to DNA. Raman spectroscopic results suggest that amsacrine has a binding preference of intercalation between AT base pairs of DNA. Minor groove binding is also observed in amsacrine-DNA complexes. These results are in good agreement with in silico investigation of amsacrine binding to DNA and thus provide detailed insight into DNA binding properties of amsacrine, which could ultimately, renders its cytotoxic efficacy.

Spectroscopic and molecular docking studies on chlorambucil interaction with DNA.

Chlorambucil (CMB) is an anticancer drug used for the treatment of variety of cancers. Structural and conformational changes associated with DNA after binding with CMB were explored using spectroscopic techniques to get insight into the mechanism of action of CMB at molecular level. Different molar ratios of CMB-DNA complex were prepared with constant DNA concentration under physiological conditions. FTIR spectroscopy, UV-visible spectroscopy, CD spectroscopy and molecular docking studies were employed to determine the binding site and binding constant of CMB with DNA. The results show CMB binds DNA through nitrogenous bases (thymine, guanine and cytosine). The binding constant was calculated to be 1.3 × 10³ M⁻¹, which suggests weak binding of CMB with DNA double helix. FTIR and CD results show that CMB do not disturb native B-conformation of DNA and it continues to remain in its B conformation even at higher concentrations of CMB. The molecular docking results are in corroboration with our experimental results and provides structural insight into the interaction site.

Binding of an indole alkaloid, vinblastine to double stranded DNA: A spectroscopic insight in to nature and strength of interaction

Vinblastine is a chemotherapeutic drug, used for the treatment of various cancers. It functions by interfering with DNA in fast growing cells and preventing them from reproducing. The present work is focused on the interaction of vinblastine with double stranded DNA in aqueous solution. Fourier transform infrared and UV-Visible absorption spectroscopy were used to analyze the interaction of vinblastine with calf-thymus DNA. FTIR analysis showed binding of vinblastine through A-T and G-C base pairs of DNA along with its phosphate backbone. UV-Vis spectroscopy results suggested the intercalation of drug in between the base pairs of DNA double helix. The binding constant estimated for vinblastine-DNA association was found to be K=1.7×10(3)M(-1). Molecular docking was performed and the results showed adenine base binding of vinblastine with DNA. Furthermore spectroscopic results revealed that formation of vinblastine-DNA complex resulted in no major change in the B-conformation of DNA.

Analysis of ovarian tumor pathology by Fourier Transform Infrared Spectroscopy

BackgroundOvarian cancer is the second most common cancer among women and the leading cause of death among gynecologic malignancies. In recent years, infrared (IR) spectroscopy has gained attention as a simple and inexpensive method for the biomedical study of several diseases. In the present study infrared spectra of normal and malignant ovarian tissues were recorded in the 650 cm-1 to 4000 cm-1 region.MethodsPost surgical tissue samples were taken from the normal and tumor sections of the tissue. Fourier Transform Infrared (FTIR) data on twelve cases of ovarian cancer with different grades of malignancy from patients of different age groups were analyzed.ResultsSignificant spectral differences between the normal and the ovarian cancerous tissues were observed. In particular changes in frequency and intensity in the spectral region of protein, nucleic acid and lipid vibrational modes were observed. It was evident that the sample-to-sample or patient-to-patient variations were small and the spectral differences between normal and diseased tissues were reproducible.ConclusionThe measured spectroscopic features, which are the spectroscopic fingerprints of the tissues, provided the important differentiating information about the malignant and normal tissues. The findings of this study demonstrate the possible use of infrared spectroscopy in differentiating normal and malignant ovarian tissues.

Rapid determination of main constituents of packed juices by reverse phase-high performance liquid chromatography: an insight in to commercial fruit drinks.

The present work reports the compositional analysis of thirteen different packed fruit juices using high performance liquid chromatography (HPLC). Vitamin C, organic acids (citric and malic) and sugars (fructose, glucose and sucrose) were separated, analyzed and quantified using different reverse phase methods. A new rapid reverse phase HPLC method was developed for routine analysis of vitamin C in fruit juices. The precision results of the methods showed that the relative standard deviations of the repeatability and reproducibility were <0.05 and <0.1 respectively. Correlation coefficient of the calibration models developed was found to be higher than 0.99 in each case. It has been found that the content of Vitamin C was less variable amongst different varieties involved in the study. It is also observed that in comparison to fresh juices, the packed juices contain lesser amounts of vitamin C. Citric acid was found as the major organic acids present in packed juices while maximum portion of sugars was of sucrose. Comparison of the amount of vitamin C, organic acids and sugars in same fruit juice of different commercial brands is also reported.

Stress degradation studies of nelfinavir mesylate by Fourier transform infrared spectroscopy.

Nelfinavir mesylate is the first nonpeptidic protease inhibitor available in pediatric formulation. In the present paper the stability of nelfinavir mesylate under different stress conditions is evaluated using Fourier transform infrared spectroscopy. The drug is subjected to thermal degradation, photodegradation, acid hydrolysis, base hydrolysis and oxidation as per ICH guidelines. Differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), X-ray diffraction (XRD) and high performance liquid chromatography (HPLC) are carried out to support the implementation of infrared spectroscopy for the stability studies of nelfinavir mesylate. Significant changes are observed in the IR spectra collected after exposing the drug to thermal radiations, acid and base hydrolysis and oxidative degradation. No change is observed in the spectra of the drug after exposing it to sunlight indicating the good photostability of nelfinavir mesylate. The results of infrared spectroscopy agree well with that of other complementary techniques as DSC, TGA, XRD and HPLC.

Plasmon induced ultrafast injection of hot electrons in Au nanoislands grown on a CdS film

Metal nanoparticle semiconductor heterostructures exhibit unique optoelectronic properties and have potential applications in energy harvesting, photodetectors, photocatalysts, and optoelectronic devices. The hot carriers are formed at the metal nanostructure semiconductor interface, and their efficient injection into the surrounding media (semiconductor) is a great challenge and understanding the physics behind the charge transfer is currently a topic of global research. Herein, we report the investigation on the hot electron injection in the Au nanoislands formed on a CdS film using ultrafast femtosecond spectroscopy. When the Au–CdS film is excited with visible light (above the band gap), a complete bleaching effect is observed. However, when it is excited with the photon energies below the band gap of the CdS film, an absorption signal is observed over a broad spectral range. The ultrafast charge transfer dynamics studied herein indicate the possibility of the plasmons formed in the Au nanoislands, which directly decay nonradiatively by injecting electrons in the conduction band of the CdS film and charge recombination with the Au nanoislands. Finally, we demonstrate the charge transfer in the metal semiconductor hybrid, which exhibits a significant alteration in the ultrafast optical properties compared to its individual components and is dependent on the excitation energy and can thus be exploited in the light harvesting devices.

Determination of Some Ethnomedicinally Important Constituents of Aegle marmelos Fruit During Different Stages of Ripening

Aim Aegle marmelos is a medicinal herb belonging to the Rutacae family. The fruit of A. marmelos at each stage of ripening is used as ethnomedicine to cure various diseases. The aim of the present study was to determine the components contributing to the medicinal value of the A. marmelos fruit at different stages of ripening.