Interactions of Janus Green B with double stranded DNA and the determination of DNA based on the measurement of enhanced resonance light scattering

A novel assay of DNA with a sensitivity at the nanogram level is proposed based on the measurement of enhanced resonance light scattering (RLS) signals resulting from the interaction of Janus Green B (JGB) with DNA. At pH 6.37 and ionic strength < 0.20, the RLS signals of JGB were greatly enhanced by DNA in the region of 300-650 nm characterized by three peaks at 416.0, 452.0 and 469.2 nm. The binding properties were examined using a Scatchard plot based on the measurement of the enhanced RLS data at 416.0 nm at a high JGB: DNA molar ratio (R > 2.22), and an aggregation mechanism of JGB in the presence of DNA at the nanogram level is proposed. Linear relationships can be established between the enhanced RLS intensity and DNA concentration in the range 0-3.5 micrograms ml-1 for both calf thymus DNA (ctDNA) and fish sperm DNA (fsDNA) if 2.0 x 10(-5) M JGB is employed. The limits of determination were 8.7 ng ml-1 for ctDNA and 9.9 ng ml-1 for fsDNA, respectively. Synthetic samples were analysed satisfactorily.     

Multi-spectroscopic method study the interaction of anti-inflammatory drug ketoprofen and calf thymus DNA and its analytical application

Interactions of the anti-inflammatory drug ketoprofen with calf thymus DNA (ctDNA) in aqueous solution have been studied by multi-spectroscopic method including resonance light scattering (RLS) technique, ultraviolet spectra (UV), (1)H NMR, etc. The characteristics of RLS spectra, the effective factors and optimum conditions of the reaction have been unequivocally investigated. Mechanism investigations have shown that ketoprofen can bind to ctDNA by groove binding and form large particles, which resulted in the enhancement of RLS intensity. In Critic acid-Na(2)HPO(4) buffer (pH=6.5), ketoprofen has a maximum peak 451.5 nm and the RLS intensity is remarkably enhanced by trace amount of ctDNA due to the interaction between ketoprofen and ctDNA. The enhancement of RLS signal is directly proportional to the concentration of ctDNA in the range of 1.20×10(-6)-1.0×10(-5) mol/L, and its detection limit (3σ) is 1.33×10(-9) mol/L. The method is simple, rapid, practical and relatively free from interference generated by coexisting substance, and was applied to the determination of trace amounts of nucleic acid in synthetic samples with satisfactory results.     

Screen anticancer drug in vitro using resonance light scattering technique

An in vitro screening model using resonance light scattering (RLS) technique with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) reagent as the reactive probe to target cancer cell was firstly developed. In this model, MTT was reduced by viable cancer cells to produce a purple formazan. Cell viability was proportional to the number of formazan induced strong light scattering signal. The inhibition rate of anticancer drug was found to vary inversely with the H₂₂-MTT system RLS intensity. So it was intuitive to see the sequence of the tumor suppressive activity of six anticancer drugs without data processing by RLS/MTT screening spectra. Compared with the traditional MTT method, this method has high sensitivity, low detection limit and quite intuitive screening results which were identical to those obtained from the MTT colorimetric assay.     

A resonance light-scattering off–on system for studies of the selective interaction between adriamycin and DNA

On the basis of the resonance light scattering (RLS) of Ag nanoparticles (AgNPs), an RLS off–on system was developed for studies of the selective interaction between adriamycin (ADM) and DNA. In this strategy, addition of ADM could induce a proportional decrease in the RLS intensity of AgNPs; this could be used to detect trace amounts of ADM with a detection limit of 12.75 ng mL⁻¹ in the range 0.021–10.0 μg mL⁻¹. Subsequently, by investigating the ability of different DNA sequences to restore the RLS intensity of the analytical systems, we found that ADM was selective to dsDNA and had an obvious preference for sequences that were rich in guanine and cytosine bases. In order to validate the results of the RLS assay, fluorescence quenching was used, and binding constants and binding numbers of each system were calculated. Compared with other methods, this RLS off–on strategy was more sensitive, fast, and reliable. It has also supplied a novel method for studying the sequence selectivity of DNA-targeted anticancer drugs and is a novel application of the RLS technique in analytical chemistry.     

Spectral studies on the interaction of Amido black 10B with DNA in the presence of cetyltrimethylammonium bromide

The interaction of Amido black 10B (AB) with DNA in basic medium was studied in the presence of cetyltrimethylammonium bromide (CTMAB) based on the measurements of resonance light scattering (RLS), UV–vis, CD spectra, and RLS imaging. The interaction has been proved to give a ternary complex of CTMAB–DNA–AB in Britton–Robinson buffer of pH 11.55, which exhibits strong negative Cotton effect at 233.3 nm and 642.8 nm, and strong RLS signals characterized at 469 nm. Experiments showed that the enhanced RLS intensities (ΔI_RLS) against the mixture of AB and CTMAB are proportional to the concentration of fish sperm DNA (fsDNA) and calf thymus DNA (ctDNA), respectively over the range of 0.03–1.0 and 0.05–1.5 μg ml⁻¹, with the limits of determination (3σ) of 7.3 ng ml⁻¹ for fsDNA and 7.0 ng ml⁻¹ for ctDNA.     

Multiplex binding modes of toluidine blue with calf thymus DNA and conformational transition of DNA revealed by spectroscopic studies

It is noteworthy to understand the details of interactions between antitumor drugs and DNA because the binding modes and affinities affect their antitumor activities. Here, The interaction of toluidine blue (TB), a potential antitumor drug for photodynamic therapy of tumor, with calf thymus DNA (ctDNA) was explored by UV–vis, fluorescence, circular dichroism (CD) spectroscopy, UV-melting method and surface-enhance Raman spectroscopy (SERS). The experimental results suggest that TB could bind to ctDNA via both electrostatic interaction and partial intercalation. The fluorescence quenching of TB by ctDNA was static and due to electron transfer from bases to the excited singlet state of TB. At low [TB]/[DNA] ratio, TB mainly partially intercalated into ctDNA resulting in the slight increase of base stacking degree; at high [TB]/[DNA] ratio, excessive TB externally stacked along the helix surface via coupling with partially intercalated ones, thereby inducing B-A transition of ctDNA. The conformational transition of DNA was confirmed by the obvious improvement of the thermal stability of ctDNA. The SERS spectra suggest that TB could partially intercalate into DNA basepairs with its ring C₁NC_1′ side buried.     

Study on bromocresol green-cetyltrimethylammonium-deoxyribonucleic acids system by resonance light scattering spectrum methods

An assay of deoxyribonucleic acids (DNA) determination, with the sensitivity at nanogram level, was established in the present study by using a common spectrofluorometer to detect the intensity of resonance light scattering (RLS). In hexamethylene tetramine (HMTA) buffer (pH 11.00), Bromocresol Green (BCG) and deoxyribonucleic acids (DNA) react with cetyltrimethylammonium bromide (CTMAB) to form large particles of three-component complex, which results in strong enhanced RLS signals characterized by three peaks at 336, 390, and 622 nm and at 336 nm that is the strongest of the three enhanced RLS peaks. Mechanistic studies showed that the enhanced RLS stems from the aggregation of BCG on DNA through the bridged and synergistic effect of CTMAB. Yeast DNA (yDNA), in the range of 0.05-0.90 ngml(-1), fish sperm DNA (fsDNA) in the range of 0.05-0.80 ngml(-1), and calf thymus DNA (ctDNA) in the range of 0.05-0.80 ngml(-1) can be determined if 2.0 x 10(-6) moll(-1) BCG was employed. The determination limit of yDNA was 12.7 ngml(-1). Three synthetic samples of yDNA were analyzed with good reproducibility.     

Resonance light scattering spectroscopy study of interaction between norfloxacin and calf thymus DNA and its analytical application

The interaction between norfloxacin and calf thymus double-stranded DNA (dsDNA) has been studied by a resonance light scattering (RLS) technique with a common spectrofluorometer. The characteristics of RLS spectra, the effective factors and optimum conditions of the reaction have been investigated. In Britton-Robinson (BR) buffer (pH 5.87), norfloxacin has a maximum peak 405.5 nm and the RLS intensity is remarkably enhanced by trace amount of calf thymus dsDNA due to the interaction between norfloxacin and dsDNA. The binding of norfloxacin to DNA forms large particles, which were characterized by RLS spectrum, scanning electron microscopy (SEM), ultraviolet-visible (UV-vis) spectrum, and fluorescence spectrum. Based on the enhanced RLS intensity, a novel method for sensitive determination of calf thymus dsDNA concentration ranging from 0.02 to 2.3 microg ml(-1) was developed. The determination limit (3 sigma) was 1.2 ng ml(-1). The method is simple, rapid, practical and relatively free from interference generated by coexisting substance, as well as much more sensitive than most of the reported methods. Three synthetic samples of ctDNA were determined with satisfactory results.     

Study of the interaction of Azur B with DNA and the determination of DNA based on resonance light scattering measurements

Based on the measurements of molecular absorption and resonance light scattering (RLS), the aggregation of Azur B (AB) was in a medium of pH ranging from 1.98 to 2.56 and ionic strength <0.12 M. The presence of double stranded DNA prompts the aggregation, resulting in enhanced RLS signals. Linear relationships were achieved between the enhanced RLS intensity at 359.7 nm and DNA concentration in the range of 0-4.5 μg ml⁻¹ for both calf thymus DNA (ctDNA) and fish sperm DNA (fsDNA) if 3.0×10⁻⁵ M AB was employed. The 3σ limits of detection were 9.3 and 8.9 ng ml⁻¹ for ctDNA and fsDNA, respectively. Five synthetic samples were analysed satisfactorily.     

Rapid and ultrasensitive electrochemical detection of circulating tumor DNA by hybridization on the network of gold-coated magnetic nanoparticles

An accurate and robust method for quantifying the levels of circulating tumor DNA (ctDNA) is vital if this potential biomarker is to be used for the early diagnosis of cancer. The analysis of ctDNA presents unique challenges because of its short half-life and ultralow abundance in early stage cancers. Here we develop an ultrasensitive electrochemical biosensor for rapid detection of ctDNA in whole blood. The sensing of ctDNA is based on hybridization on a network of probe DNA modified gold-coated magnetic nanoparticles (DNA-Au@MNPs). This DNA-Au@MNPs biosensor can selectively detect short- and long-strand DNA targets. It has a broad dynamic range (2 aM to 20 nM) for 22 nucleotide DNA target with an ultralow detection limit of 3.3 aM. For 101 nucleotide ctDNA target, a dynamic range from 200 aM to 20 nM was achieved with a detection limit of 5 fM. This DNA-Au@MNPs based sensor provides a promising method to achieve 20 min response time and minimally invasive cancer early diagnosis.

This study introduces a new electrochemical sensing strategy for the rapid detection of circulating tumor DNA (ctDNA) from whole blood in combination with a network of DNA-Au@MNPs with high sensitivity and excellent selectivity.     

Investigation on the interaction between anthracyclines and DNA in the presence of paclitaxel by resonance light scattering technique

We have developed a method to investigate the interaction between DNA-targeted anthracyclines and DNA in the presence of the drug paclitaxel. It is based on resonance light scattering (RLS) and on the finding that anthracyclines when bound to DNA undergo a dramatic enhancement in their RLS intensities, while paclitaxel does not display such an effect. However, the RLS intensities of the anthracyclines-DNA associates are remarkably enhanced again on addition of paclitaxel. UV-visible spectra reveal interactions between paclitaxel and anthracyclines, but no reaction between paclitaxel and DNA. Consequently, paclitaxel, though not DNA-targeted, can improve the DNA-binding capabilities of anthracyclines. Binding constants between anthracyclines and DNA, and improved efficiency of paclitaxel on the DNA-binding capabilities of anthracyclines were calculated. The DNA binding constants of doxorubicin, epirubicin, and mitoxantrone, respectively, are 4.53?×?10⁵?L?mol^?1, 6.05?×?10⁵?L?mol^?1, and 9.47?×?10⁵?L?mol^?1. The improved values in presence of paclitaxel are 78%, 47% and 19%. We also have investigated the effects of drug concentrations and the order of adding the drugs. Displacement studies (using methylene blue as a competitive agent) provided additional information on the mechanisms of the interaction between paclitaxel and anthracyclines.

Study on spectroscopic characterization of Cu porphyrin/Co porphyrin and their interactions with ctDNA

The luminescence behaviors of Cu(II) meso-tetrakis (4-N-methyl-pyridiniumyl) porphyrin (Cu(II)TMPyP) and Co(II) meso-tetrakis (4-N-methyl-pyridiniumyl) porphyrin (Co(II)TMPyP) are investigated, and their interactions with calf thymus DNA (ctDNA) are studied by UV-vis, fluorescence and resonance light scattering (RLS) and based on the changes of UV-vis spectra, fluorescence and RLS spectra, the intrinsic binding constants of Cu(II)TMPyP/Co(II)TMPyP with ctDNA are obtained in the case of phosphate buffer solution (pH 7.2), respectively. According to the experimental results, it can be inferred that the interaction model of Cu(II)TMPyP with ctDNA is intercalative binding, while Co(II)TMPyP is the long-range assembly on the molecular surface of ctDNA.     

Electrochemical Determination of the Effect of Caffeic Acid onto the Interaction between Idarubicin and DNA by Single‐use Disposable Electrodes

Now it is well known, antioxidant may affect the interaction of anticancer drugs and DNA. This study aims to investigate the interaction between Idarubicin and DNA and the effect of caffeic acid on this interaction. Disposable, inexpensive, easy handle electrodes were used in this study to investigate the interaction of idarubicin and DNA electrochemically. Idarubicin (IDR) is an anthracyline antitumor antibiotic, which used against one or more types of leukemia. Electrochemical behaviour of IDR was investigated by using cyclic voltammetry. The interaction between anticancer drug, IDR and calf thymus double‐stranded DNA (ctdsDNA) was investigated by using differential pulse voltammetry (DPV) technique in the absence and presence of caffeic acid.     

Interaction between insulin and calf thymus DNA, and quantification of insulin and calf thymus DNA by a resonance Rayleigh scattering method

The interaction of insulin with calf thymus deoxyribonucleic acid (ctDNA) leads to a complex that displays remarkably enhanced resonance Rayleigh scattering (RRS). The complex and its formation were investigated by atomic force microscopy and by absorption, fluorescence and circular dichroism spectroscopies. We show that the Tyr B16, Tyr B26 and Phe B24 amino acids near the active center (Phe B25) were influenced by the interaction, whereas Tyr A14, Tyr A19 and Phe B1 (which are located far away from the active center) were less influenced. The interaction provide a way in the quantitation of both ctDNA and insulin with high sensitivity. When ctDNA is used as a probe to quantify insulin, the detection limit (3σ) is 6.0?ng?mL^-1. If, inversely, insulin is used as a probe to quantify ctDNA, the detection limit (3σ) is 7.2?ng?mL^-1. The analysis of synthetic DNA samples and an insulin infection sample provided satisfactory results.

Micro-determination of nucleic acids with a simple probe manganese chloride based on the fine enhanced resonance light scattering

Manganese chloride can form large particles with nucleic acids by electrostatic forces, which results in strong enhancement of resonance light scattering (RLS) signals. Based on this phenomenon, a novel and very simple assay of DNA was established. The work conditions have been investigated including the concentration of probe, the acidity of solution, the effect of ionic strength and the selectivity. In acidic solution, the enhanced RLS intensity at 389.5 nm was proportional to the concentration of nucleic acids in the range 0.05-10.0 microg ml(-1) for both ctDNA and fsDNA and 1.0-10.0 microg ml(-1) for yRNA. The limits of detection (LOD, 3sigma) were 0.17, 0.13 and 0.53 ng ml(-1) for ctDNA, fsDNA and yRNA, respectively. Synthetic samples were determined satisfactorily.     

Cetyltrimethylammonium bromide sensitized resonance light-scattering of nucleic acid--Pyronine B and its analytical application

This is the first report on the determination of nucleic acids with Pyronine B (PB) sensitized by cetyltrimethylammonium bromide (CTMAB) with resonance light-scattering (RLS) technique. Under the experimental conditions (1 x 10(-5) mol l(-1) PB, 1 x 10(-5) mol l(-1) CTMAB, pH 7.4, at room temperature, ionic strength 0.02 mol l(-1) NaCl), the interaction of PB with DNA sensitized by CTMAB results in enhanced RLS signals at 328 and 377 nm in the enhanced regions. It was found that the enhanced RLS intensity at 328 nm was proportional to the concentration of DNA in the suitable ranges. The linear range of this assay is 0.0-1.2 microg ml(-1) for calf thymus, 0.0-0.8 microg ml(-1) for fish sperm DNA (fsDNA), and 0.04-1.4 microg ml(-1) for yeast RNA, respectively. The detection limits (3 sigma) are 6.1 ng ml(-1) for calf thymus DNA (ctDNA), 11.2 ng ml(-1) for fish sperm DNA, and 8.6 ng ml(-1) for yeast RNA, respectively. Six synthetic samples were determined satisfactorily. This method is simple, rapid and the dye is inexpensive and stable.     

A resonance light scattering quenching system for studying DNA sequence recognition of actinomycin D

The DNA sequence recognition study of DNA-targeted anticancer drugs is a theoretical basis for improving the selectivity of anticancer drugs. With the high synergy effect of cocoamidopropyl hydroxy sulfobetaine (HSB), a resonance light scattering (RLS) quenching system for DNA sequence recognition studies of actinomycin D (ACTD) was developed in this contribution. By the strategy, DNA sequence selectivity as well as the recognition mechanisms of ACTD was systematically investigated. The results suggested that ACTD had the selectivity to single-stranded DNA (ssDNA) with an equilibrium constant (K(RLS)) of 12.4 mmol mg(-1). Also it had a preference for Guanine and Cytosine bases with a K(RLS) of 6.69 L mmol(-1). The selectivity mechanism between ACTD and DNA was also well discussed with the help of UV-Vis absorption spectroscopy. Compared with other methods, the RLS quenching system has the advantages of reliability and speediness, and it avoids complex modification processes and is a better bionic system for the above research. Results obtained from this work would supply a theoretical basis for improving anticancer activity and designing similar anticancer drugs.     

Study on the resonance light-scattering spectrum of lysozyme-DNA/CdTe nanoparticles system

The interactions of lysozyme and calf thymus DNA (ctDNA) or thioglycolic acid (TGA) modified CdTe nanoparticles in aqueous solution have been studied by resonance light-scattering (RLS) spectroscopy. At pH 7.2 Britton-Robinson (BR) buffer solution and pH 7.4 phosphate buffered saline (PBS), the RLS signals of ctDNA and TGA modified CdTe nanoparticles were greatly enhanced by lysozyme in the region of 220-750 nm characterized by the peak around 306 and 353 nm, respectively. Under optimal conditions, the increase of RLS intensity of the two systems is proportional to the concentration of lysozyme. The linear range is 0.1-25 microg/ml for the lysozyme-ctDNA system, and 0.2-10.7 microg/ml for the lysozyme-TGA modified CdTe nanoparticles system. The detection limit is 0.041 microg/ml for the lysozyme-ctDNA system, and 0.083 microg/ml for the lysozyme-TGA modified CdTe nanoparticles system, respectively. Meanwhile lysozyme can also be used as a probe to determine the ctDNA. The increase of RLS intensity of the system is also proportional to the concentration of ctDNA. The linear range is 0.078-13 microg/ml. The detection limit is 0.024 microg/ml. Three kinds of samples were analyzed with satisfactory results.     

吖啶橙-细胞DNA荧光抑制法筛选抗癌药物

在DNA荧光法‘’‘初步筛选抗癌药物的实验中,漠化乙锭（EB）是常用的荧光探针,其本身荧光板弱,但能与双链DNA发生专一性插入作用,使荧光大幅度地增强．但EB不能进入完整的细胞膜［’j．对陕橙（AO）常用于细胞内DNA和RNA的定量测定,并能用于活细胞染色;AO与核酸的结合方式有2种：一种是在嵌入核酸双链的碱基对之间形成AO－DNA复合物,另一种是与单链核酸的磷酸发生静电间相互作用,形成AO－RNA复合物．如用蓝光激发,前者发射峰波长为530urn的绿色荧光,后者发射峰波长为640urn的红色荧卅’‘．本文提出一种对细胞周期非…     

Resonance light scattering spectroscopy study of interaction between gold colloid and thiamazole and its analytical application

In this paper, we used resonance light scattering (RLS) spectroscopy to study the interaction between thiol-containing pharmaceutical-thiamazole and gold colloid. At pH 5.2, the resonance light scattering spectrum of gold nanoparticles has a maximum peak at 555 nm and the RLS intensity is enhanced by trace amount of thiamazole due to the interaction between thiamazole and gold colloid. The binding of colloidal gold to thiamazole results in ligand-induced aggregation of colloidal gold, which was characterized by RLS spectrum, ultraviolet-visible (UV-Vis) spectrum, and transmission electron microscopy (TEM). Based upon the study, we proposed a highly sensitive, gold colloid-based assay using RLS spectrum to detect pharmaceuticals for the first time. The mechanism of binding interaction between Au colloid and thiamazole was also discussed.