Evaluation of Nitric Oxide Fluctuation Via a Fast, Responsive Fluorescent Probe in Idiopathic Pulmonary Fibrosis Cells and Mice Models.

Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, and fatal interstitial pneumonia with unknown pathogenesis. Early diagnosis and therapeutic intervention are essential for improving the prognosis of patients with IPF. The level of nitric oxide upregulates in the alveoli of IPF patients, which is correlated with the severity of the disease. Herein, we report a fluorescent probe DCM-nitric oxide (NO) to detect IPF by monitoring the concentration changes of NO. This probe displays a fast response time and a good linear response to NO in vitro. Fluorescence imaging experiments with probe DCM-NO revealed that the level of intracellular NO increases in the pulmonary fibrosis cells and IPF mice models. Probe DCM-NO displayed a strong red fluorescence in IPF mice models. However, a declining fluorescence was evidenced in the OFEV-treated IPF mice, implying that DCM-NO is capable of evaluating the therapeutic effects on IPF. Thus, probe DCM-NO can quickly predict the progression of pulmonary fibrosis at an early stage and thus help improve the effective treatment.

Integrating Target-Triggered Aptamer-Capped HRP@Metal-Organic Frameworks with a Colorimeter Readout for On-Site Sensitive Detection of Antibiotics.

Colorimetric analytical strategies exhibit great promise in developing on-site detection methods for antibiotics, while substantial recent research efforts remain problematic due to dissatisfactory sensitivity. Taking this into account, we develop a novel colorimetric sensor for in-field detection of antibiotics by using aptamer (Apt)-capped and horseradish peroxidise (HRP)-embedded zeolitic metal azolate framework-7 (MAF-7) (Apt/HRP@MAF-7) as target recognition and signal transduction, respectively. With the substrate 3,3',5,5'-tetramethylbenzidine (TMB)-impregnated chip attached on the lid, the assay can be conveniently operated in a tube and reliably quantified by a handheld colorimeter. Hydrophilic MAF-7 can not only prevent HRP aggregation but also enhance HRP activity, which would benefit its detection sensitivity. Besides, the catalytic activity of HRP@MAF-7 can be sealed through assembling with Apt and controllably released based on the bioresponsivity via forming target-Apt complexes. Consequently, a significant color signal can be observed owing to the oxidation of colorless TMB to its blue-green oxidized form oxTMB. As a proof-of-concept, portable detection of streptomycin was favorably achieved with excellent sensitivity, which is superior to most reported methods and commercial kits. The developed strategy affords a new design pattern for developing on-site antibiotics assays and immensely extends the application of enzyme embedded metal-organic framework composites.

l-Pyroglutamic Acid-Modified CdSe/ZnS Quantum Dots: A New Fluorescence-Responsive Chiral Sensing Platform for Stereospecific Molecular Recognition.

Stereoselective recognition of amino acids is extremely important due to its high chirality-dependent interactions and physiological activities in life activities. We herein report a novel functionalized chiral fluorescent nanosensor prepared from surface modification of CdSe/ZnS quantum dots (QDs) with pyroglutamic acid derivatives, which could serve as a chiral recognition module for fluorescence detection of chiral molecules. The sensor exhibited a unique stereoselective fluorescence response to histidine (His), glutamate (Glu), and dihydroxyphenylalanine (Dopa) and had preferable response performance to l-enantiomers. The enantiomeric fluorescence difference ratios of His, Glu, and Dopa enantiomers were 3.90, 3.40, and 2.49, respectively. The mechanism for the enantiomeric fluorescence recognition was systematically studied through a fluorescence spectrum, fluorescence life, and density functional theory (DFT) calculation. Presumably, the different hydrogen bonding capacity of the chiral recognition module with two enantiomers mainly contributed to the difference in fluorescence signals. As a result, a broader application of the pyroglutamic acid derivative-coated QDs as a fluorescence-responsive chiral sensing platform for enantiomeric detection would be expected.

Preparative isolation and purification of 12 main antioxidants from the roots of Polygonum multiflorum Thunb. using high-speed countercurrent chromatography and preparative HPLC guided by 1,1'-diphenyl-2-picrylhydrazyl-HPLC.

A hyphenated strategy by off-line coupling of 1,1'-diphenyl-2-picrylhydrazyl-high-performance liquid chromatography, high-speed countercurrent chromatography, and preparative high-performance liquid chromatography was established to screen and separate antioxidants from ethyl acetate fraction of the roots of Polygonum multiflorum. Under the targeted guidance of 1,1'-diphenyl-2-picrylhydrazyl-high-performance liquid chromatography experiment, 12 compounds were identified as potential antioxidants and readily isolated by high-speed counter-current chromatography and preparative high-performance liquid chromatography. Ultraviolet spectroscopy, mass spectrometry, and 1 H NMR spectroscopy were employed to identify their structures, which were assigned as gallic acid (1, 6.2 mg, 98.28%), catechin (2, 8.8 mg, 90.69%), epicatechin (3, 4.1 mg, 96.71%), polydatin (4, 5.3 mg, 94.91%), 2,3,5,4'-tetrahydroxy stilbene-2-Ο-ß-D-glucoside (5, 20.2 mg, 95.23%), piceatannol (6, 5.3 mg, 96.85%), rutin (7, 5.4 mg, 97.92%), resveratrol (8, 5.2 mg, 96.94%), isorhapontigenin (9, 11.4 mg, 94.81%), hyperoside (10, 9.7 mg, 98.52%), rhein (11, 4.9 mg, 97.46%), and emodin (12, 8.2 mg, 95.74%). Notably, compounds 6 and 9 were isolated from Polygonum multiflorum for the first time. In addition, antioxidant activity of compounds 1-12 were evaluated, and compounds 1-8 and 10 exhibited stronger antioxidant activity than ascorbic acid (positive control). These results indicated that the proposed method is a highly efficient strategy to screen and isolate antioxidants from complex natural products.

Rapid screening and identification of antioxidants in the leaves of Malus hupehensis using off-line two-dimensional HPLC-UV-MS/MS coupled with a 1,1'-diphenyl-2-picrylhydrazyl assay.

The leaves of Malus hupehensis have a strong antioxidant activity and are commonly consumed as a healthy tea. However, detailed information about its antioxidants is incomplete. Herein, we developed an effective strategy based on combining off-line two-dimensional high-performance liquid chromatography with ultraviolet and tandem mass spectrometry detection with a 1,1'-diphenyl-2-picrylhydrazyl assay to rapidly screen and identify the antioxidants from the leaves of M. hupehensis. In the orthogonal two-dimensional liquid chromatography system, a Venusil HILIC column was used for the first dimension, while a Universil XB-C18 column was installed in the second dimension. As a result, 32 antioxidants, including ten dihydrochalcones, two flavanones, nine flavonols, four flavones, and seven phenolic acids were tentatively identified, out of which 23 compounds, as far as we know, were isolated and characterized from the leaves of M. hupehensis for the first time. To the best of our knowledge, this is the first systematic investigation of the antioxidants from the leaves of M. hupehensis. The results indicated that the proposed method is an efficient technique to rapidly investigate antioxidants, especially for coeluted and minor compounds in a complex system.

In situ growth of ZIF-8 on gold nanoparticles/magnetic carbon nanotubes for the electrochemical detection of bisphenol A.

We herein report a facile and scalable strategy for the fabrication of a metal-organic framework (MOF) based composite by in situ growing ZIF-8 on gold nanoparticle (AuNP) loaded magnetic carbon nanotubes (mCNTs). AuNPs were firstly loaded on PEI (polyethylenimine) modified mCNTs by electrostatic forces, and then AuNPs/mCNTs were encapsulated into the ZIF-8 frame through in situ self-assembling of zinc ions and 2-methylimidazole. The morphology, spectroscopy and structural properties of the AuNP/mCNT@ZIF-8 nanocomposites were systematically characterized. The conductivity-strain tests revealed that the in situ insertion of AuNPs/mCNTs in ZIF-8 could not only shorten the electron transfer distance between active sites and mCNTs, but also increase the dispersion of mCNTs, which would benefit the electron and mass transfer. Besides, by adopting the AuNP/mCNT@ZIF-8 nanocomposite-modified glassy carbon electrode (GCE) as the working electrode, a novel electrochemical sensor was successfully developed for the detection of bisphenol A (BPA). A linear range of BPA detection from 1 µM to 100 µM with a limit of detection of 690 nM was favorably obtained. Moreover, the developed sensor exhibited satisfactory reproducibility and superior stability with excellent anti-interference ability, and was successfully applied in the detection of BPA in real samples.

Integrating amino acid oxidase with photoresponsive probe: A fast quantitative readout platform of amino acid enantiomers.

Low-cost, high-throughput, broadly useful photoresponsive enantiomeric excess (ee) sensing of amino acids remains challenging to date. Herein, based on the selective oxidation reaction of amino acid oxidase (AAO) to amino acid enantiomers (D/L-AA) and the oxidation reaction of substrate (H2O2) with aromatic boronic ester, we put forward a photoresponsive strategy for the determination of D/L-AA at a certain concentration. In this scheme, the substrate H2O2 produced by the enzyme-catalyzed reaction was determined by sensitive fluorescent and colorimetric response of ethyl-3-(3-(benzothiazol-2-yl)-5-methyl-2-((4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)oxy)phenyl)-2-cyanoacrylate (HBT-PB) to reflect the enantiomeric content at a certain concentration. The photoresponsive probe HBT-PB was readily available and inexpensive with sensitive long-wavelength red fluorescence and colorimetric light response to H2O2, the detection limit (LOD) was estimated as 2.91 µM. The operation of the sensing method was simple and data collection and processing are straightforward. The practicability of the scheme was favorably confirmed by accurate and scientific analysis of methionine and Dopa samples. As a result, the scheme was not only suitable for high-throughput screening but also adaptable to low-cost and sensitive RGB colorimetric analysis platform (LOD of methionine and Dopa was calculated as 9.23 µM and 8.34 µM respectively) with modern plate readers, and possessed extremely high enantioselectivity and wide applicability which benefited from the specificity and efficiency of enzyme catalytic reaction.

Rapid and visual detection of aflatoxin B1 in foodstuffs using aptamer/G-quadruplex DNAzyme probe with low background noise.

Contamination of foods by aflatoxin B1 (AFB1) is a common serious problem. To improve the efficiency of AFB1 detection, this study aims to develop a sensitive aptasensor for detection of AFB1 in food samples based on the catalytic effect of aptamer/G-quadruplex DNAzyme probe. The resulting reassembly of this probe in the presence of hemin and K+ catalyzes the generation of fluorescent 2,3-diaminophenazine (DAP) from o-phenylenediamine (OPD). Interestingly, we first found that the high background induced by the superfluous hemin can be effectively suppressed with the aid of sequential adsorption and magnetic separation by magnetic oxidized multiwall carbon nanotubes (Fe3O4@oMWCNTs). This aptasensor exhibits a high sensitivity toward AFB1 with a detection limit of 0.02 ng/mL. The assay also shows higher selectivity for AFB1 compared to other reported agents and can be employed to detect AFB1 in foodstuffs, which might find broad practical applications in other food contaminants determination.

Intelligent Platform for Simultaneous Detection of Multiple Aminoglycosides Based on a Ratiometric Paper-Based Device with Digital Fluorescence Detector Readout.

A digital fluorescence detector (DFD), a handheld fluorescence detection device, can convert the fluorescence signal of samples into the corresponding fluorescer concentration. Herein, by adopting a DFD as the readout, a novel intelligent platform was developed based on a ratiometric paper-based device (RPD) for multiple aminoglycoside detection. There are five layers and four parallel channels contained in the designed RPD, functioning as reagent storage, fluidic path control and signal processing, respectively. The rationale of this design lies in the fact that aptamer/graphitic carbon nitride nanosheet (Apt/g-C3N4 NS) modified layers can catalyze o-phenylenediamine to fluorescent 2,3-diaminophenazine (DAP) in the presence of H2O2. When Apt was removed from nanosheets via the Apt-target reaction, the peroxidase-like activity would be decreased, thus decreasing the production of DAP. All the changes of the fluorescence DAP signal can be read out using a portable DFD. Based on the DFD signal change related to the concentration of the target, a quantitative reaction platform was established. Furthermore, the sample flow and Apt-target reaction time can be reasonably regulated using the H2O2-cleavable hydrophobic compound modified layer placed between the target recognition region and detection region. Then, the practicality of this platform was verified through realizing sensitive analysis of streptomycin, tobramycin, and kanamycin simultaneously. Overall, with merits including portability and ease of operation, the platform shows great potential in on-site simultaneous detection of multiple targets, especially in resource-limited settings.

Development of a "Dual Gates" Locked, Target-Triggered Nanodevice for Point-of-Care Testing with a Glucometer Readout.

Developing a facile and sensitive sensing platform is of importance for point-of-care testing (POCT). Herein, a sensitive and portable POCT platform based on "dual gates" aminated magnetic mesoporous silica nanocomposites (AMMS) bearing polydopamine (PDA)-aptamer (Apt) two-tier shells, as a novel nanodevice, is designed for target detection through a target-triggered glucose (GO) release from AMMS with personal glucometer (PGM) readout. In the absence of target, GO can be firmly captured in pores by the designed "dual gates", which would decrease the high background signal of this system and ensure the accuracy of the detection results. Upon the introduction of the target molecules under acidic conditions (pH 5.5), the subsequent PDA self-degradation and the specific Apt-target reaction can cause the departure of "dual gates" and the opening of pores to release the loaded GO molecules, which could be quantitatively monitored by a portable PGM. It has been demonstrated that such POCT platform shows high sensitivity and excellent selectivity for aflatoxin B1 (AFB1) detection, accompanied by the well-presented reproducibility and stability. Importantly, this sensing platform was further validated by assaying contaminated samples, where the obtained results were well matched with that by HPLC. Regarding the features of portability, high sensitivity, and high throughput detection, the developed platform might find wide applications in POCT.

Single-stranded DNA modified protonated graphitic carbon nitride nanosheets: A versatile ratiometric fluorescence platform for multiplex detection of various targets.

Facile and cost-effective detection of multiple targets is essential for a variety of applications ranging from life sciences to environmental monitoring. Here, we report a versatile ratiometric fluorescence platform for multiple detection of various targets based on the conjugation of single-stranded DNA (ssDNA) with protonated graphitic carbon nitride nanosheets (Pg-C3N4 NSs). We demonstrate that intrinsic peroxidase-like activity of Pg-C3N4 NSs is enhanced by conjugating with ssDNA, and thus the oxidation of substrate o-phenylenediamine (OPD) is promoted in the presence of H2O2. The oxidation product 2,3-diaminophenazine (DAP) can deliver a new fluorescence signal at 564 nm, and concurrently quench the intrinsic fluorescence of conjugates ssDNA/Pg-C3N4 NSs at 443 nm upon excitation at 370 nm. The transformation of fluorescence provides us a novel strategy for ratiometric fluorescence-based analytical sensing. Taking ssDNA as the target-recognition element of the conjugates ssDNA/Pg-C3N4 NSs, we favorably present ratiometric fluorescence detection of various targets including heavy metal ions (Hg2+) and biomolecules (Aflatoxin B1 (AFB1) and adenosine triphosphate (ATP)) in real samples by varying the ssDNA sequences. The present work provides a new strategy to develop facile methods for quantitative determination of various analytes and uncovers an innovative horizon for Pg-C3N4 NSs-based sensing platform fabrication.

Organocatalytic, Enantioselective, Polarity-Matched Ring-Reorganization Domino Sequence Based on the 3-Oxindole Scaffold.

A one-pot squaramide-catalyzed enantioselective ring-reorganization domino sequence (Michael addition/intramolecular ring-opening/lactamization) of 3-hydroxyoxindole and methyleneindolinone, which can be readily derived from 3-oxindole, has been established in this work. As a result, novel polycyclic quinolinone-spirooxindoles bearing three contiguous chiral centers were efficiently and step-economically assembled under mild conditions in high yields (up to 97%) with excellent enantioselectivities (up to >99% ee) and moderate to good diastereoselectivities (up to >95:5 dr).

Novel S, N-doped carbon quantum dot-based "off-on" fluorescent sensor for silver ion and cysteine.

In this work, sulfur and nitrogen co-doped carbon dots (S,N-CQDs) as highly selective fluorescent probe for silver ion (Ag+) and cysteine (Cys) detection were designed and synthesized directly from citric acid and thiamine hydrochloride via a one-step hydrothermal protocol in 63.8% quantum yield. This probe enabled selective detection of Ag+ with a linear range of 0-10 and 10-250µM and a limit of detection of 0.40µM with respect to the variation in fluorescence induced by target concentration and electron-transfer from S,N-CQDs to Ag+. Furthermore, S,N-CQDs/Ag+ fluorescence can be effectively recovered by virtue of a specific reaction of Cys with silver ion. This fluorescence "turn-on" protocol was applied to determine Cys with two linear ranges of 0-10 and 10-120µM as well as a detection limit of 0.35µM. The corresponding cell experiments were also performed, indicating that the prepared S,N-CQDs possessed low cytotoxicity and good biocompatibility. Ultimately, the practicality and viability of this fluorescent probe were demonstrated through the analysis of silver ion in real river water and human serum samples.

4-Mercaptophenylboronic acid-modified spirally-curved mesoporous silica nanofibers coupled with ultra performance liquid chromatography-mass spectrometry for determination of brassinosteroids in plants.

Herein, for the first time, thiol-functionalized mesoporous silica (mSiO2-SH) nanofibers with a spirally-curved twisted hexagonal morphology were synthesized via a simple one-pot protocol. 4-Mercaptophenylboronic acids (4-MPBA) were attached onto the mSiO2-SH nanofibers via disulfide bond, serving as boronate affinity sorbent to selectively capture brassinosteroids (BRs) from plant extract. The resulting BRs-MPBA derivatives were easily eluted from the sorbent by cleaving the disulfide bond, which was subsequently subjected to ultra performance liquid chromatography-mass spectrometry (UPLC-MS) analysis. Thus, the in situ extraction/derivatization/desorption method coupled with UPLC-MS was established for the fast, sensitive and selective detection of BRs in plant tissues. Finally, based on the developed method, endogenous BRs were successfully detected in leaf of H. lupulus L., silique of A. thaliana, and panicle of O. sativa L.

Sensitive surface enhanced Raman spectroscopy (SERS) detection of methotrexate by core-shell-satellite magnetic microspheres.

An effective approach for fabricating core-shell-satellite magnetic microspheres was developed to form a sub-10nm positively charged mesoporous silica (mSiO2) interlayer, which was applied to enrich samples and create sufficient hotspots between the inner Fe3O4@Ag core and outer assembled gold nanoparticles (AuNPs) satellites. Based on the prepared Fe3O4@Ag/mSiO2/AuNPs (FASA) as SERS substrates, a sensitive and label-free detection of methotrexate in serum was successfully realized. The positively charged mSiO2 interlayer exhibited strong capability in enriching methotrexate through the electrostatic interaction. The intraparticle plasmonic coupling between the inner Ag shell and the outside AuNPs satellites, and magnetism-induced aggregation greatly enhanced the SERS activity of methotrexate. The diluted serum containing different concentrations of methotrexate was employed as real samples and the minimum detected concentration of methotrexate was as low as 1nM with a linear response range from 1 to 200nM. This sensitive and label-free SERS detection of methotrexate based on FASA offers great potential for practical applications in medicine and biotechnology.

Multispectroscopic Investigation of the Interaction Between two Ruthenium(II) Arene Complexes of Curcumin Analogs and Human Serum Albumin.

The interaction between two ruthenium(II) arene complexes of curcumin analogs and human serum albumin (HSA) was systematically investigated by multispectroscopic techniques. The fluorescence spectral results indicated that two complexes quenched the intrinsic fluorescence of HSA through static quenching mode. The quenching constants and the corresponding thermodynamic parameters at different temperatures were calculated. The binding interactions of two complexes with HSA resulted in the complex formation of complex-HSA, and the van der Waals interactions and hydrogen bond interactions played major roles in the complex stabilization. The distances between HSA and two complexes were obtained according to fluorescence resonance energy transfer theory. The site competitive replacement experiments illustrated that two complexes mainly bounded with HSA on site I. The results of synchronous fluorescence spectra, three-dimensional fluorescence spectra, FT-IR spectra, and circular dichroism spectra indicated that the secondary structure of HSA was changed at the present of two complexes. The results of mass spectrometry further validated the binding interaction and the binding number between two complexes and HSA.

Study on the molecular interaction of graphene quantum dots with human serum albumin: combined spectroscopic and electrochemical approaches.

Graphene quantum dots (GQDs) have attracted great attention in biological and biomedical applications due to their super properties, but their potential toxicity investigations are rarely involved. Since few studies have addressed whether GQDs could bind and alter the structure and function of human serum albumin (HSA), the molecular interaction between GQDs and HSA was systematically characterized by the combination of multispectroscopic and electrochemical approaches. GQDs could quench the intrinsic fluorescence of HSA via static mode. The competitive binding fluorescence assay revealed that the binding site of GQDs was site I of HSA. Some thermodynamic parameters suggested that GQDs interacted with HSA mainly through van der Waals interactions and hydrogen bonding interactions, and protonation might also participate in the process. As further revealed by FT-IR spectroscopy and circular dichroism technique, GQDs could cause the global and local conformational change of HSA, which illustrated the potential toxicity of GQDs that resulted in the structural damage of HSA. Electrochemical techniques demonstrated the complex formation between GQDs and HSA. Our results offered insights into the binding mechanism of GQDs with HSA and provided important information for possible toxicity risk of GQDs to human health.

Thermodynamic investigation of interaction between [(η6-p-cymene) RuII(acetone-N4-phenylthiosemicarbazone)Cl]Cl anticancer drug and human serum albumin: spectroscopic and electrochemical studies.

In this contribution, the interaction between [(η (6)-p-cymene)Ru(II)(acetone-N (4)-phenylthiosemicarbazone)Cl]Cl (Ru-TSC) anticancer drug and human serum albumin (HSA) was investigated by spectroscopic and electrochemical techniques. The fluorescence spectra results indicated that Ru-TSC anticancer drug could quench the intrinsic fluorescence of HSA through dynamic quenching mode. The calculated corresponding activation energy of the interaction between Ru-TSC anticancer drug and HSA was 35.62 kJ mol(-1). The distance between HSA and Ru-TSC anticancer drug was obtained according to fluorescence resonance energy transfer. The results of synchronous fluorescence spectra, three-dimensional fluorescence spectra, Fourier transform infrared spectroscopy (FTIR) spectra, and circular dichroism (CD) spectra indicated that the microenvironment and the conformation of HSA were all changed in the presence of Ru-TSC anticancer drug. The results of cyclic voltammetry further validated the interaction between Ru-TSC and HSA. These results indicated that the biological activity of HSA was affected by Ru-TSC anticancer drug dramatically.

A sensitive quantum dots-based "OFF-ON" fluorescent sensor for ruthenium anticancer drugs and ctDNA.

In this contribution, a simple and sensitive fluorescent sensor for the determination of both the three ruthenium anticancer drugs (1 to 3) and calf thymus DNA (ctDNA) was established based on the CdTe quantum dots (QDs) fluorescence "OFF-ON" mode. Under the experimental conditions, the fluorescence of CdTe QDs can be effectively quenched by ruthenium anticancer drugs because of the surface binding of these drugs on CdTe QDs and the subsequent photoinduced electron transfer (PET) process from CdTe QDs to ruthenium anticancer drugs, which render the system into fluorescence "OFF" status. The system can then be "ON" after the addition of ctDNA which brought the restoration of CdTe QDs fluorescence intensity, since ruthenium anticancer drugs broke away from the surface of CdTe QDs and inserted into double helix structure of ctDNA. The fluorescence quenching effect of the CdTe QDs-ruthenium anticancer drugs systems was mainly concentration dependent, which could be used to detect three ruthenium anticancer drugs. The limits of detection were 5.5 × 10(-8) M for ruthenium anticancer drug 1, 7.0 × 10(-8) M for ruthenium anticancer drug 2, and 7.9× 10(-8) M for ruthenium anticancer drug 3, respectively. The relative restored fluorescence intensity was directly proportional to the concentration of ctDNA in the range of 1.0 × 10(-8) M ∼ 3.0 × 10(-7) M, with a correlation coefficient (R) of 0.9983 and a limit of detection of 1.1 × 10(-9) M. The relative standard deviation (RSD) for 1.5 × 10(-7) M ctDNA was 1.5% (n = 5). There was almost no interference to some common chemical compounds, nucleotides, amino acids, and proteins. The proposed method was applied to the determination of ctDNA in three synthetic samples with satisfactory results. The possible reaction mechanism of CdTe QDs fluorescence "OFF-ON" was further investigated. This simple and sensitive approach possessed some potential applications in the investigation of interaction between drug molecules and DNA.