Binding of triclosan to human serum albumin: insight into the molecular toxicity of emerging contaminant

Purpose

The interaction between triclosan (TCS) and human serum albumin (HSA) was investigated in order to obtain the binding mechanism, binding constant, the type of binding force, the binding distance between the donor and acceptor, and the effect of TCS on the conformation change of HSA.

Methods

A HSA solution was added to the quartz cell and then titrated by successive addition of TCS. The fluorescence quenching spectra and synchronous spectra were recorded with the excitation and emission slits of the passage of band set at 10 and 20 nm. Three-dimensional fluorescence spectra of HSA were recorded before and after the addition of TCS. The capillary electrophoresis was conducted with the pressure injection mode at 0.5 psi for 5 s, separation under 25 kV, and detection at 214 nm.

Results

Fluorescence data indicated the fluorescence quenching of HSA by TCS was static quenching, and the quenching constants (K _a ) were 1.14?×?10⁵, 8.75?×?10⁴, 6.67?×?10⁴, and 5.00?×?10⁴ at 293, 298, 303, and 309 K, respectively. The thermodynamic parameters, enthalpy change (??H) and entropy change (??S) for the interaction were calculated to be ?37.9 kJ mol^?1 and 32.6 J?mol^?1 K^?1. The binding distance between TCS and tryptophan residues of HSA was obtained to be 1.81 nm according to F??rster nonradioactive energy transfer theory. The UV-Vis absorption spectroscopy, the synchronous fluorescence spectroscopy, three-dimensional fluorescence spectroscopy, and circular dichroism spectroscopy revealed the alterations of HSA secondary structure in the presence of TCS. Finally, the interaction between TCS and HSA was further confirmed by capillary electrophoresis.

Conclusions

TCS was bound to HSA to form the TCS-HSA complex, with the binding distance of 1.81 nm. Hydrophobic interaction and hydrogen bond were dominated in the binding. TCS could change the secondary conformation of HSA. This work provides an insight into noncovalent interaction between emerging pollutants and protein, helping to elucidate the toxic mechanism of such pollutants.     

Potential toxicity of amphenicol antibiotic: binding of chloramphenicol to human serum albumin

Antibiotics are widely used in daily life but their abuse has posed a potential threat to human health. To evaluate the toxicity of chloramphenicol (CAP) at the protein level, the interaction between CAP and human serum albumin (HSA) was investigated by fluorescence, Ultraviolet–visible (UV–Vis) absorption, Fourier transform infrared (FT-IR) spectroscopy and molecular docking methods. Fluorescence data revealed that the fluorescence quenching of HSA by CAP was the result of the formation of CAP–HSA complex, and the binding constant was determined to be 3.196?×?10⁴ L mol^?1 at 310 K. The thermodynamic determination indicated that the interaction was driven by enthalpy change and entropy change together, where the multiple hydrogen bonds (CAP and the residues Arg 222 and His 242 of HSA) and van der Waals forces were the dominant binding force. The site marker competition revealed that CAP bound into sub-domain IIA of HSA. The binding of CAP induced the drastic reduction in α-helix conformation and the significant enhancement in β-sheet conformation of HSA. Molecular docking study further confirmed the binding mode obtained by experimental study. This work provides a new quantitative evaluation method for antibiotics to cause the protein damage.     

Interaction of tebuconazole with bovine serum albumin: determination of the binding mechanism and binding site by spectroscopic methods

Abstract

This study investigates the interaction between tebuconazole and bovine serum albumin (BSA) in a physiological buffer (pH = 7.4) using the fluorescence quenching method to obtain the apparent binding constants (K) and number of binding sites (n) in the interaction between tebuconazole and BSA. The results revealed that tebuconazole can quench the intrinsic fluorescence of BSA through a static quenching procedure. It also shows that the thermodynamic parameters of enthalpy change (ΔH) and entropy change (ΔS) are negative, indicating that the interaction of tebuconazole with BSA is mainly driven by van der Waals forces and hydrogen bonds. The process of binding was a spontaneous process in which Gibbs free energy change was negative. The distance of r between the donor (BSA) and acceptor (tebuconazole) was calculated to be 0.68?nm based on Forster’s non-radiative energy transfer theory. Analysis of synchronous fluorescence, three-dimensional fluorescence and circular dichroism (CD) spectra demonstrates that tebuconazole can induce conformational changes of BSA.     

Investigations on the interaction of the phototoxic alkaloid coralyne with serum albumins

The interaction of the phototoxic alkaloid coralyne with bovine and human serum albumins (BSA, HSA) was investigated. Absorbance and fluorescence quenching experiments revealed the formation of strong complexes. Based on the binding parameters calculated from Stern-Volmer quenching method, coralyne has higher affinity to BSA (∼10⁵ M⁻¹) compared to HSA (∼10⁴ M⁻¹). Forster resonance energy transfer studies showed that the specific binding distances between Trp (donor) of the proteins and coralyne (acceptor) were 2.95 and 3.10 nm, respectively. The bindings were favored by negative enthalpy and a stronger favorable entropy contribution. The heat capacity values for binding to BSA and HSA were similar, indicating the involvement of similar molecular forces in the complexation. Competitive binding experiments using site markers demonstrated that coralyne binds to site I (subdomain IIA) of both proteins. The secondary structure of the proteins was altered, suggesting a small but definitive partial unfolding on complexation.     

Interaction of diuron to human serum albumin: Insights from spectroscopic and molecular docking studies

This investigation was undertaken to determine the interaction of diuron with human serum albumin (HSA) was studied by monitoring the spectral behavior of diuron-HSA system. The fluorescence of HSA at 340 nm excited at 230 nm was obviously quenched by diuron due to dynamic collision and the quenching constant was of the order of 10⁴ L mol^?1 at 310 K. However, no fluorescence quenching was observed when excited at 280 nm. Thermodynamic investigations revealed that the combination between diuron and HSA was entropy driven by predominantly hydrophobic interactions. The binding of diuron induced the drastic reduction in α-helix conformation and the significant enhancement in β-turn conformation of HSA. In addition, both sites marker competition study and molecular modeling simulation evidenced the binding of diuron to HSA primarily took place in subdomain IIIA (Sudlow's site II).     

Pesticide binding and urea-induced controlled release applications with calixarene naphthalimide molecules by host–guest complexation

ABSTRACT

Three novel calix[4]arene molecule-based 1,8 naphthalimide fluoroionophore for the selective determination of kesoxim-methyl were synthesized and used in pesticide binding studies. The possible interaction between pesticides and fluorescent calix[4]arene molecules was monitored by UV/Vis absorption and fluorescence spectroscopy. When compared the studied pesticides, kesoxim-methyl was strongly quenched the fluorescence intensity of upper rim-modified calix[4]arene. UV and fluorescence titration experiments were also studied to determine both the quenching mechanism and stoichiometric ratio consisted in complex formation. Furthermore, pesticide release experiments were also performed with a fertilizing agent as urea by using fluorescence spectroscopy technique.     

Study on the mechanism of action between dimethyl phthalate and herring sperm DNA at molecular level

Dimethyl phthalate (DMP), a typical phthalic acid ester, is widespread in the environment and causes extensive concern due to its adverse effects on human health. To understand the genotoxicity of DMP at molecular level, the toxic interaction of DMP with herring sperm (hs) deoxyribonucleic acid (DNA; hs-DNA) was investigated in vitro under simulated physiological conditions using multi-spectroscopic techniques and a molecular modeling method. The results of Ultraviolet-Visible absorption spectroscopy, fluorescence emission spectroscopy, and circular dichroism spectra indicated that DMP interacts with hs-DNA in a groove-binding mode that changes the double helical structure of DNA. The binding constant and the number of binding sites calculated from the fluorescence quenching data were 565.718 L mol^?1 and 0.7872, respectively. A molecular modeling study revealed that DMP tends to bind with DNA in the A-T-rich regions of minor groove and that hydrogen bonding and van der Waals forces play main roles in the interaction. This research can help to elucidate the mechanism of DMP toxicity in vivo.     

Kinetics and equilibrium properties of the biosorption of Cu2+ by algae

The purpose of this study was to examine the kinetics and equilibrium properties of freshwater algae with Cu²⁺. This was a model system to explore using algae as biosensors for water quality. Methods included making luminescence measurements (fluorescence) and copper ion-selective electrode (CuISE) measurements vs. time to obtain kinetic data. Results were analyzed using a pseudo-first-order model to calculate the rate constants of Cu²⁺ uptake by algae: k _p(Cu?Calgae)?=?0.0025?±?0.0006?s^?1 by CuISE and k _p(Cu?Calgae)?=?0.0034?±?0.0011?s^?1 by luminescence. The binding constant of Cu?Calgae, K _Cu?Calgae, was 1.62?±?0.07?×?10⁷?M^?1. Fluorescence results analyzed using the Stern?CVolmer relationship indicate that algae have two types of binding sites of which only one appears to affect quenching. The fluorescence-based method was found to be able to detect the reaction of algae with Cu²⁺ quickly and at a detection limit of 0.1?mg?L^?1.     

Characterization on the toxic mechanism of two fluoroquinolones to trypsin by spectroscopic and computational methods

Abstract

Ciprofloxacin (CPFX) and enrofloxacin (ENFX), two of the most widely used fluoroquinolones (FQs), pose a great threat to humans and the ecosystem. In this study, the toxic mechanisms between the two FQs and trypsin were evaluated by means of multiple spectroscopic methods, as well as molecular docking. During the fluorescence investigations, both FQs quenched the intrinsic fluorescence of trypsin effectively, which was due to the formation of moderately strong complexes (mainly through van der Waals forces and hydrogen bonds). The binding of two FQs not only caused the conformational and micro-environmental changes of trypsin, but also changed its molecular activity; shown by the UV–Visible absorption spectroscopy, synchronous fluorescence spectroscopy, and functional tests. The established methods in this work can help to comprehensively understand the transport of FQs in the human body.     

Investigation of metal ions binding of humic substances using fluorescence emission and synchronous‐scan spectroscopy

Abstract

The binding site interactions of IHSS humic substances, Suwannee River Humic Acid, Suwannee River Fulvic Acid, Nordic Fulvic Acid, and Aldrich Humic Acid with various metals ions and a herbicide, methyl viologen were investigated using fluorescence emission and synchronous‐scan spectroscopy. The metal ions used were, Fe(III), Cr(III), Cr(VI), Pb(II), Cu(II) and Ni(II). Stern‐Volmer constants, K_SV for these quenchers were determined at pH 4 and 8 using an ionic strength of 0.1M. For all four humic substances, and at both pH studied, Fe(III) was found to be the most efficient quencher. Quenching efficiency was found to be 3–10 times higher at pH 8. The bimolecular quenching rate constants were found to exceed the maximum considered for diffusion controlled interactions, and indicate that the fluorophore and quencher are in close physical association. Synchronous‐scan spectra were found to change with pH and provided useful information on binding site interactions between humic substances and these quenchers.     

Elemental analysis of the edible fruit of Carpobrotus dimidiatus (from Kwazulu-Natal,South Africa) and the influence of soil quality on its elemental uptake

Abstract

Carpobrotus dimidiatus is an indigenous South African medicinal plant species from the Aizoaceae family that bears edible fruit that is consumed for nutritional value. In this study, the elemental distribution in C. dimidiatus fruit and growth soil from fifteen sites in KwaZulu-Natal (South Africa) was determined along with soil pH, soil organic matter and cation exchange capacity, to assess for nutritional value and the effect of soil quality on elemental uptake. The results showed elemental concentrations in fruit to be in decreasing order of Ca (6235–32755?mg kg^?1) > Mg (2250–5262?mg kg^?1) > Fe?>?Mn?>?Zn (20.9–50.6?mg kg^?1) > Cu (3.83–20.6?mg kg^?1) > Pb?>?Cr?>?Cd?>?As?～?Co?～?Ni?～?Se and no potential health risk due to metal toxicity from average consumption. For sites that had high levels of Cd and Pb, bioaccumulation occurred from atmospheric deposition. Concentrations of elements in soil were found to be in decreasing order of Fe (1059–63747?mg kg^?1) > Ca (1048–41475?mg kg^?1) > Mg?>?Mn (9.76–174?mg kg^?1) > Cr (1.55–135?mg kg^?1) > Zn (0.76–58.2?mg kg^?1) > Se?>?Cu?>?Ni?>?Pb?>?Co?>?As?～?Cd with no evidence of heavy metal contamination. This study revealed that the plant inherently controlled uptake of essential elements according to physiological needs and that the concentrations of essential elements in the fruit could contribute positively to the diet.     

Uptake and accumulation of antimicrobials,triclocarban and triclosan,by food crops in a hydroponic system

Commonly used in personal care products, triclocarban (TCC) and triclosan (TCS) are two chemicals with antimicrobial properties that have recently been recognized as environmental contaminants with the potential to adversely affect human health. The objective of the study described herein was to evaluate the potential of food crops to uptake TCC and TCS. Eleven food crops, grown in hydroponic nutrient media, were exposed to a mixture of 500 μg L^?1 TCC and TCS. After 4 weeks of exposure, roots accumulated 86–1,350 mg kg^?1 of antimicrobials and shoots had accumulated 0.33–5.35 mg kg^?1 of antimicrobials. Translocation from roots to shoots was less than 1.9 % for TCC and 3.7 % for TCS, with the greatest translocation for TCC observed for pepper, celery, and asparagus and for TCS observed for cabbage, broccoli, and asparagus. For edible tuber- or bulb-producing crops, the concentrations of both TCC and TCS were lower in the tubers than in the roots. Exposure calculations using national consumption data indicated that the average exposure to TCC and TCS from eating contaminated crops was substantially less than the exposure expected to cause adverse effects, but exceeded the predicted exposure from drinking water. Exposure to antimicrobials through food crops would be substantially reduced through limiting consumption of beets and onions.     

Differential aggregation of polystyrene and titanium dioxide nanoparticles under various salinity conditions and against multiple proteins types

The interaction of nanoplastics (NPls) and engineered nanoparticles (ENPs) with organic matter and environmental pollutants is particularly important. Therefore, their behavior should be investigated under the different salinity conditions, mimicking rivers and coastal environments, to understand this phenomenon in those areas. In this work, we analyzed the elementary characteristics of polystyrene-PS (unmodified surface and modified with amino or carboxyl groups) and titanium dioxide-TiO2 nanoparticles. The effect of salinity on their colloidal properties was studied too. Also, the interaction with different types of proteins (bovine serum albumin-BSA and tilapia proteins), as well as the formation of the BSA corona and its effect on the colloidal stability of nanoparticles, were evaluated. The morphology and dispersion of sizes were more uniform in unmodified-surface PS-NPs (70.5?±?13.7 nm) than in TiO2-NPs (131.2?±?125.6 nm). Likewise, Rama spectroscopy allowed recognizing peaks associated with the PS phenyl group aromatic ring in unmodified-surface PS-NPs (621, 1002, 1582, and 1602 cm^?1). For TiO2-NPs, the data suggest belonging to the tetragonal form, also known as rutile (445, 610 cm^?1). The elevation of salinity dose-dependently decreased NP colloid stability, with more significant variation in the PS-NPs compared to TiO2-NPs. The organic matter is also involved in this phenomenon, differentially as a function of time compared to its absence (unmodified-surface PS-NPs 30 psu/TOC 5 mgL^?1/24 h: 2876.6?±?378.03 nm; unmodified-surface PS-NPs 30 psu/24 h: 2133?±?49.57 nm). In general, the TiO2-NPs demonstrated greater affinity with all proteins tested (0.066 g/L). It was observed that morphology, size, and surface chemical modification intervene in a relevant way in the interaction of the nanoparticles with bovine serum albumin (unmodified-surface PS-NPs 298 K: 6.08E⁺⁰²; 310 K: 6.63E⁺⁰²; TiO2-NPs 298 K: 8.76E⁺⁰²; 310 K: 1.05E⁺⁰³ L mol^?1) and tilapia tissues proteins (from blood, gills, liver, and brain). Their morphology and size also determined the protein corona formation and the NPs’ agglomeration. These findings can provide references during knowledge transfer between NPls and ENPs.

     

Quantification of the interactions between Ca,Hg and extracellular polymeric substances (EPS) of sludge

The interactions between metals (Ca²⁺ and Hg²⁺) and extracellular polymeric substances (EPS) extracted from the aerobic and anaerobic sludge in wastewater treatment reactors were investigated using a combination of zeta potential measurement and 3-dimensional excitation–emission matrix (EEM) fluorescence spectroscopy with parallel factor (PARAFAC) analysis. Results show that Ca²⁺ had no substantial effects on the EEM fluorescence spectra of the EPS, but their zeta potentials increased with the increasing Ca²⁺ dosage. However, Hg²⁺ had a significant effect on the EEM fluorescence spectra of the EPS, while their zeta potentials seemed not to be affected by the dose of Hg²⁺. The interactions between Hg²⁺ and EPS were elucidated using the fluorescence quenching with PARAFAC analysis, while the interactions between Ca²⁺ and EPS were evaluated by the zeta potential technique. The binding constants for Hg²⁺ and EPS were two orders of magnitude higher than those for Ca²⁺ and EPS, suggesting that the binding mechanisms between Ca²⁺ and EPS were different from those between Hg²⁺ and EPS. The results might be useful for understanding the roles of EPS in bacterial self-protection against heavy metals and the aggregate formation mechanisms through ionic bridging interactions.     

The complexation of Cu(II) by anthropogenic fulvic acids extracted from composted urban and livestock wastes

Abstract

The fulvic acid (fua) fractions of two samples of composted solid wastes [urban (urfua) and livestock (lsfua) wastes], commercialized to be used in agriculture as organic correctives or fertilizers, were analyzed for their affinity towards Cu(II) at pH=6. Molecular fluorescence spectroscopy (synchronous mode) was used to monitor the quenching caused by the complexation upon addition of Cu(II) to fua. Spectral data were preprocessed by a chemometric self‐modeling mixture analysis method (SIMPLISMA) to detect the number of different types of fluorescent binding sites that exist in each fua, their spectra and the corresponding quenching profiles [fluorescence intensity as function of the total Cu(II) concentration]. From the analysis of the quenching profiles, the amount of binding sites (Cl) and the corresponding conditional stability constants (K') were calculated. Both fua samples have approximately Cl = 0.21 mmol/g and the logarithms of K’ are 4.21(3) and 4.51(8), respectively for urfua and lsfua. The differences detected between these fua samples and those extracted from natural soils can be attributed mainly to the relatively small humification extent of the present anthropogenic fua samples.     

Selected chloro and bromo derivatives of triclosan—syntheses and their occurrence in Canadian sewage and biosolid samples

The occurrence of triclosan (TCS), a general purpose antibacterial agent contained in numerous consumer and personal-care products, in the aquatic environment is well known. To a lesser degree, the formation of chlorinated and brominated derivatives of TCS during the chlorination of the antibacterial has also been reported. Presumably due to the lack of authentic standards, very few reports have been published on the levels of these halogenated TCSs in the environment. For this purpose, we have synthesized six selected halogenated derivatives of TCS, namely, 3-Cl-, 5-Cl-, 3,5-Cl₂-, 3-Br-, 5-Br-, and 3,5-Br₂- TCSs, with supporting ¹H-NMR (nuclear magnetic resonance) and GC–MS (gas chromatography–mass spectrometry) data for their structural assignments. Using these model compounds together with sensitive analytical methods, we were able to identify and quantify the above compounds together with their precursor compound TCS in Canadian municipal wastewater and biosolid samples for the first time. While detected in all influent (range from 1.4 to 24.1 ng L^?1) and biosolid (range from 7.7 to 274 ng g^?1) samples, the concentrations of these chlorinated TCS were generally from 100- to 1,000-fold lower than TCS in the same sample. Even lower levels (<20 ng/g in 85 % of the results) of brominated TCS were found in biosolids, and they were mostly undetected in sewage.     

Organic and elemental carbon associated to PM10 and PM2.5 at urban sites of northern Greece

Organic carbon (OC) and elemental carbon (EC) concentrations, associated to PM₁₀ and PM_2.5 particle fractions, were concurrently determined during the warm and the cold months of the year (July–September 2011 and February–April 2012, respectively) at two urban sites in the city of Thessaloniki, northern Greece, an urban-traffic site (UT) and an urban-background site (UB). Concentrations at the UT site (11.3?±?5.0 and 8.44?±?4.08 14 μg m^?3 for OC₁₀ and OC_2.5 vs. 6.56?±?2.14 and 5.29?±?1.54 μg m^?3 for EC₁₀ and EC_2.5) were among the highest values reported for urban sites in European cities. Significantly lower concentrations were found at the UB site for both carbonaceous species, particularly for EC (6.62?±?4.59 and 5.72?±?4.36 μg m^?3 for OC₁₀ and OC_2.5 vs. 0.93?±?0.61 and 0.69?±?0.39 μg m^?3 for EC₁₀ and EC_2.5). Despite that, a negative UT-UB increment was frequently evidenced for OC_2.5 and PM_2.5 in the cold months possibly indicative of emissions from residential wood burning at the urban-background site. At both sites, cconcentrations of OC fractions were significantly higher in the cold months; on the contrary, EC fractions at the UT site were prominent in the warm season suggesting some influence from maritime emissions in the nearby harbor area. Secondary organic carbon, being estimated using the EC tracer method and seasonally minimum OC/EC ratios, was found to be an appreciable component of particle mass particularly in the cold season. The calculated secondary contributions to OC ranged between 35 and 59 % in the PM₁₀ fraction, with relatively higher values in the PM_2.5 fraction (39–61 %). The source origin of carbonaceous species was investigated by means of air parcel back trajectories, satellite fire maps, and concentration roses. A local origin was mainly concluded for OC and EC with limited possibility for long range transport of biomass (agricultural waste) burning aerosol.     

Determination of glyphosate residue in maize and rice using a fast and easy method involving liquid chromatography–mass spectrometry (LC/MS/MS)

A fast and easy method was developed for the determination of glyphosate in maize and rice by using liquid chromatography triple quadrupole mass spectrometry with a Dionex Ion Pack column and phosphate buffer mobile phase. Samples were extracted with an acidified methanol solution. An isotope-labeled internal standard was added to the sample before extraction to ensure accurate tracking and quantification. The method’s performance was evaluated through a series of assessments to determine the accuracy, precision, linearity, matrix effect, limit of detection (LOD), and limit of quantification (LOQ). The mean recoveries for both matrices were within 70–105% at three fortification levels, including the LOQ. The precision for replicates was <20% (RSD%) for both matrices. Good linearity (R²=0.9982) was obtained over the concentration range of 0.01–1.5?mg kg^?1. The LOD was determined to be 0.002?mg kg^?1 for rice and 0.004?mg kg^?1 for maize. The LOQ was 0.01?mg kg^?1 for both maize and rice. Due to its versatility, the proposed method could be considered useful for the determination of glyphosate in cereals in routine analysis.     

Biopesticide and formulation processes based on starch industrial wastewater fortified with soybean medium

Abstract

The aim of this study was to produce Bacillus thuringiensis-based biopesticide using starch-producing industry wastewater (SIW) fortified with soybean medium and optimize the formulated product using different adjuvants. This study was necessary as low endotoxin concentration is obtained in formulated biopesticide when SIW alone is used as fermentation medium. The fermentation runs were conducted using SIW alone and SIW fortified with 25% soybean (w/v) medium in 2000?L and 150?L bioreactor, respectively. SIW supplemented with soybean medium showed an increase in cell count (from 1.95?×?10⁸ to 1.65?×?10⁹ CFU mL^–1), spore synthesis (from 1.5?×?10⁸ to 1.35?×?10⁹ CFU mL^–1) and endotoxin concentration (from 436 to 1170?μg mL^–1) when compared to SIW medium alone. The fermented broth was concentrated using continuous centrifugation and adjuvants were added for biopesticide formulation in order to enhance its resistance against UV rays and rainfastness. Entomotoxicity of the formulation produced using fermented broth of SIW fortified with soybean (38,000?IU μL^–1) was higher than that obtained by SIW medium alone (21,000?IU μL^–1), commercial biopesticide Foray 76B (20,000?IU μL^–1) and Btk sander’s (12,500?IU μL^–1).