A study on the interaction between cadmium and α-chymotrypsin and the underlying mechanisms.

Because cadmium might interact with proteins and, thus, exert toxicity in organisms, it is vital to understand the molecular mechanism of the interaction between cadmium and biologically relevant proteins as well as the structural and functional changes in these proteins. In this study, the interaction between α-chymotrypsin (α-ChT) and cadmium chloride (CdCl2 ) was investigated by performing enzyme activity determinations, multispectroscopic measurements, isothermal titration calorimetry, and molecular docking studies. It was demonstrated that CdCl 2 binds to α-ChT mainly via electrostatic forces with (21.0 ± 0.982) binding sites, leading to the increase of α-helix and the decrease of ß-sheet. The interaction between CdCl 2 and α-ChT loosened the protein skeleton and increased the molecular volume of α-ChT. CdCl 2 first binds to the interface of α-ChT and then interacts with the key residues His 57 or Asp 102 or both in the active sites, leading to the activity inhibition of α-ChT under the exposure of high CdCl 2 concentrations.

Development of an in Vitro Autocatalytic Self-Replication System for Biosensing Application.

Molecular self-replication is a fundamental function of all living organisms with the capability of templating and catalyzing its own synthesis, and it plays important roles in prebiotic chemical evolution and effective synthetic machineries. However, the construction of the self-replication system in vitro remains a great challenge and its application for biosensing is rare. Here, we demonstrate for the first time the construction of an in vitro enzymatic nucleic acid self-replication system and its application for amplified sensing of human 8-oxoguanine DNA glycosylase (hOGG1) based on autocatalytic self-replication-driven cascaded recycling amplification. In this strategy, hOGG1 excises 8-oxoguanine (8-oxoG) to unfold the hairpin substrate, activating the autonomous biocatalytic process with molecular beacons (MBs) as both the fuels for producing nucleic acid templates and the generators for signal output, leading to the continuous replication of biocatalytic nucleic acid templates and the repeated cleavage of MBs for an enhanced fluorescence signal. This strategy exhibits an extremely low detection limit of 4.3 × 10-7 U/µL and a large dynamic range of 5 orders of magnitude from 1 × 10-6 to 0.05 U/µL. Importantly, it can be applied for the detection of enzyme kinetic parameters, the screening of hOGG1 inhibitors, and the quantification of hOGG1 activity in even 1 single lung cancer cell, providing a new approach for biomedical research and clinical diagnosis.

The Effect of Ultraviolet Radiation on the Chemical Bath Deposition of Bis(thiourea) Cadmium Chloride Crystals and the Subsequent CdS Obtention.

In this work, the effects on the preparation of bis(thiourea) cadmium chloride crystals when illuminated with ultraviolet (UV) light at a wavelength of 367 nm using the chemical bath deposition technique are studied comparatively. Two experiments are performed to make a comparison: one without UV light and the other with the aid of UV light. Both experiments are performed under equal conditions, at a temperature of 343 K and with a pH of 3.2. The precursors used are cadmium chloride (CdCl2) and thiourea [CS(NH2)2], which are dissolved in 50 mL of deionized water with an acidic pH. In this experiment, the interaction of electromagnetic radiation is sought at the moment the chemical reaction is carried out. The results demonstrate the existence of an interaction between the crystals and the UV light; the UV light assistance causes crystal growths in an acicular shape. Also, the final product obtained is cadmium sulfide and shows no evident difference when synthesized with or without the use of UV light.

Epirubicin-calf thymus DNA interaction: a comprehensive investigation using molecular docking, spectroscopy and fluorescent quantum dots.

Reviewing the mode of interaction between this kind of active pharmaceutical ingredients and DNA has received much more attention in current years. Anthracycline drugs such as Epirubicin are frequently used in cancer treatment for breast cancer treatment. In the present study, the Epirubicin -calf thymus DNA interaction was investigated by using spectroscopic, fluorimetric and molecular docking methods. Water-soluble quantum dots (QDs) with nanometric particle size fabricated and characterized by transmission electron microscope and photon correlation spectroscopy. The binding constant value and the free energy change for this interaction were obtained to be 3.00×106 M-1 and -42.26 kJ mol-1, using the spectroscopic method and docking investigations, respectively. Additionally, fluorescent thioglycolic acid-capped CdTe QDs were used for investigation of EPI and DNA interaction. Epirubicin as a quencher quenched the fluorescence of CdTe QDs after electrostatic adsorption on the surface of QDs. With the addition of DNA, EPI will be desorbed from the surface of CdTe QDs, inserted into the DNA. Subsequently, fluorescence changes of QDs were used for calculation of binding constant value, which was in good agreement with that obtained by the spectroscopic method. By the comparison of the achieved results, the intercalation mode of interaction between Epirubicin and DNA proved.

Cadmium stress assessment based on the electrocardiogram characteristics of zebra fish (Danio rerio): QRS complex could play an important role.

The electrocardiogram (ECG) of zebra fish (Danio rerio) expresses cardiac features that are similar to humans. Here we use sharp microelectrode measurements to obtain ECG characteristics in adult zebra fish and analyze the effects of cadmium chloride (CdCl2) on the heart. We observe the overall changes of ECG parameters in different treatments (0.1 TU, 0.5 TU and 1.0 TU CdCl2), including P wave, Q wave, R wave, S wave, T wave, PR interval (atrial contraction), QRS complex (ventricular depolarization), ST segment, and QT interval (ventricular repolarization). The trends of the ECG parameters showed some responses to the concentration and exposure time of CdCl2, but it was difficult to obtain more information about the useful indicators in water quality assessment depending on tendency analysis alone. A self-organizing map (SOM) showed that P values, R values, and T values were similar; R wave and T wave amplitude were similar; and most important, QRS value was similar to the CdCl2 stress according to the classified data patterns including CdCl2 stress (E) and ECG components based on the Ward linkage. It suggested that the duration of QRS complex was related to environmental stress E directly. The specification and evaluation of ECG parameters in Cd2+ pollution suggested that there is a markedly significant correlation between QRS complex and CdCl2 stress with the highest r (0.729) and the smallest p (0.002) among all ECG characteristics. In this case, it is concluded that QRS complex can be used as an indicator in the CdCl2 stress assessment due to the lowest AIC data abased on the linear regression model between the CdCl2 stress and ECG parameters.

Involvement of Notch1 signaling in malignant progression of A549 cells subjected to prolonged cadmium exposure.

Cadmium exposure is known to increase lung cancer risk, but the underlying molecular mechanisms in cadmium-stimulated progression of malignancy are unclear. Here, we examined the effects of prolonged cadmium exposure on the malignant progression of A549 human lung adenocarcinoma cells and the roles of Notch1, hypoxia-inducible factor 1α (HIF-1α), and insulin-like growth factor 1 receptor (IGF-1R)/Akt/extracellular signal-regulated kinase (ERK)/p70 S6 kinase 1 (S6K1) signaling pathways. Exposing A549 cells to 10 or 20 µm cadmium chloride (CdCl2) for 9-15 weeks induced a high proliferative potential, the epithelial-mesenchymal transition (EMT), stress fiber formation, high cell motility, and resistance to antitumor drugs. Of note, the CdCl2 exposure increased the levels of the Notch1 intracellular domain and of the downstream Notch1 target genes Snail and Slug. Strikingly, siRNA-mediated Notch1 silencing partially suppressed the CdCl2-induced EMT, stress fiber formation, high cell motility, and antitumor drug resistance. In addition, we found that prolonged CdCl2 exposure induced reduction of E-cadherin in BEAS-2B human bronchial epithelial cells and antitumor drug resistance in H1975 human tumor-derived non-small-cell lung cancer cells depending on Notch1 signaling. Moreover, Notch1, HIF-1α, and IGF-1R/Akt/ERK/S6K1 activated each other to induce EMT in the CdCl2-exposed A549 cells. These results suggest that Notch1, along with HIF-1α and IGF-1R/Akt/ERK/S6K1 signaling pathways, promotes malignant progression stimulated by prolonged cadmium exposure in this lung adenocarcinoma model.

Molecular mechanism investigation of the neutralization of cadmium toxicity by transferrin.

Cadmium adversely affects the biological function of the liver. Transferrin might be involved in the detoxification system of cadmium. However, owing to the lack of investigation of the molecular mechanism of cadmium conjugating to transferrin, the role of transferrin in cadmium detoxification in the liver and how transferrin undergoes conformational and functional changes upon cadmium binding are not clear. In this article, we demonstrated the potential role of transferrin in the protection of the mouse primary hepatocytes against cadmium toxicity. After the incubation of hepatocytes with 10 and 100 µM CdCl2, pretreatment with transferrin significantly attenuated the reduction of cell viability in comparison with the samples treated with CdCl2 alone. Furthermore, a detailed molecular mechanism investigation of the interaction of CdCl2 with transferrin was reported using biophysical methods. Multi-spectroscopic measurements showed that CdCl2 formed complexes with transferrin and caused structural and conformational changes of transferrin. Isothermal titration calorimetry measurements revealed that transferrin has two classes of binding sites with different binding constants for CdCl2 binding. Hydrophobic forces and electrostatic forces are the major driving forces of the interaction. Preferred specific binding sites on transferrin were identified by dialysis experiments, molecular docking studies and molecular dynamics simulations. Upon low CdCl2 concentration exposure, no content of iron was released from transferrin because CdCl2 preferentially binds to the surface of transferrin molecules. Upon higher CdCl2 concentration exposure, the release of iron content from transferrin was observed due to the interaction of CdCl2 with the key residues around iron binding sites.

Biomineralization of a Cadmium Chloride Nanocrystal by a Designed Symmetrical Protein.

We have engineered a metal-binding site into the novel artificial ß-propeller protein Pizza. This new Pizza variant carries two nearly identical domains per polypeptide chain, and forms a trimer with three-fold symmetry. The designed single metal ion binding site lies on the symmetry axis, bonding the trimer together. Two copies of the trimer associate in the presence of cadmium chloride in solution, and very high-resolution X-ray crystallographic analysis reveals a nanocrystal of cadmium chloride, sandwiched between two trimers of the protein. This nanocrystal, containing seven cadmium ions lying in a plane and twelve interspersed chloride ions, is the smallest reported to date. Our results indicate the feasibility of using rationally designed symmetrical proteins to biomineralize nanocrystals with useful properties.

Apoptotic effect of novel Schiff based CdCl2(C14H21N3O2) complex is mediated via activation of the mitochondrial pathway in colon cancer cells.

The development of metal-based agents has had a tremendous role in the present progress in cancer chemotherapy. One well-known example of metal-based agents is Schiff based metal complexes, which hold great promise for cancer therapy. Based on the potential of Schiff based complexes for the induction of apoptosis, this study aimed to examine the cytotoxic and apoptotic activity of a CdCl2(C14H21N3O2) complex on HT-29 cells. The complex exerted a potent suppressive effect on HT-29 cells with an IC50 value of 2.57 ± 0.39 after 72 h of treatment. The collapse of the mitochondrial membrane potential and the elevated release of cytochrome c from the mitochondria to the cytosol indicate the involvement of the intrinsic pathway in the induction of apoptosis. The role of the mitochondria-dependent apoptotic pathway was further proved by the significant activation of the initiator caspase-9 and the executioner caspases-3 and -7. In addition, the activation of caspase-8, which is associated with the suppression of NF-κB translocation to the nucleus, also revealed the involvement of the extrinsic pathway in the induced apoptosis. The results suggest that the CdCl2(C14H21N3O2) complex is able to induce the apoptosis of colon cancer cells and is a potential candidate for future cancer studies.

The Effect of Cadmium on COX-1 and COX-2 Gene, Protein Expression, and Enzymatic Activity in THP-1 Macrophages.

The aim of this study was to examine the effects of cadmium in concentrations relevant to those detected in human serum on cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) expression at mRNA, protein, and enzyme activity levels in THP-1 macrophages. Macrophages were incubated with various cadmium chloride (CdCl2) solutions for 48 h at final concentrations of 5 nM, 20 nM, 200 nM, and 2 µM CdCl2. The mRNA expression and protein levels of COXs were analyzed with RT-PCR and Western blotting, respectively. Prostaglandin E2 (PGE2) and stable metabolite of thromboxane B2 (TXB2) concentrations in culture media were determined using ELISA method. Our study demonstrates that cadmium at the highest tested concentrations modulates COX-1 and COX-2 at mRNA level in THP-1 macrophages; however, the lower tested cadmium concentrations appear to inhibit COX-1 protein expression. PGE2 and TXB2 production is not altered by all tested Cd concentrations; however, the significant stimulation of PGE2 and TXB2 production is observed when macrophages are exposed to both cadmium and COX-2 selective inhibitor, NS-398. The stimulatory effect of cadmium on COXs at mRNA level is not reflected at protein and enzymatic activity levels, suggesting the existence of some posttranscriptional, translational, and posttranslational events that result in silencing of those genes' expression.

pH dependence of cadmium-contaminated drinking water on the development of cardiovascular injury in Wistar rats.

The purpose of our study was to investigate the effect of water pH in the genesis of cardiovascular injury caused by cadmium poisoning. For this study, 90 male Wistar rats were used, divided into six groups: A, 15 rats that received 400 mg/l cadmium chloride (CdCl2) in drinking water at a neutral pH of 7.0; B, 15 rats that received CdCl2 (400 mg/l) in drinking water at an acidic pH of 5.0; C, 15 rats that received CdCl2 (400 mg/l) in drinking water at a basic pH of 8.0; D, 15 rats that received water at an acidic pH of 5.0; E, 15 rats that received water at a basic pH of 8.0; and F, 15 rats that received water at a neutral pH of 7.0. All animals were euthanized after 6 months. We collected the heart and aorta from each rat for microscopic analysis. No microscopic changes were observed in the hearts. In the aorta, fatty streaks appeared in a large proportion of animals in groups A (50 %) and B (46 %), but fatty streaks appeared in a smaller minority of animals in groups C (15.3 %), D (0 %), E (7 %), and F (13.3 %) (p < 0.05). Cadmium exposure caused the development of fatty streaks in the aorta of animals and the exposure to this metal in basic pH decreased the formation of these lesions.

Superposition model study of Cr3+ doped tetra methyl ammonium cadmium chloride.

The zero field splitting parameter D of Cr(3+) doped in tetra methyl ammonium cadmium chloride (TMCC) is calculated with perturbation formula using microscopic spin Hamiltonian theory and crystal field parameters from superposition model. The theoretically calculated ZFS parameter for Cr(3+) in TMCC single crystal is compared with the experimental value obtained by electron paramagnetic resonance (EPR). The local structure distortion is considered to obtain the crystal field parameters. The theoretical study gives the ZFS parameter D similar to that from experiment. However, calculation considering small distortion in local structure around Cr(3+) gives better agreement with the experimental value of ZFS parameter.

Synthesis, crystal structure and thermal decomposition of the new cadmium selenite chloride, Cd4(SeO3)2OCl2.

A synthetic study in the Cd-Se-O-Cl system led to formation of the new oxochloride compound Cd4(SeO3)2OCl2 via solid state reactions. The compound crystallizes in the orthorhombic space group Fmmm with cell parameters a = 7.3610(3) Å, b = 15.4936(2) Å, c = 17.5603(3) Å, Z = 8, S = 0.969, F(000) = 2800, R = 0.0185, Rw = 0.0384. Single crystal X-ray data were collected at 293 K. The crystal structure can be considered as layered and the building units are distorted [Cd(1)O6] octahedra, distorted [Cd(2)O8] cubes, irregular [Cd(3)O4Cl2] polyhedra and SeO3E trigonal pyramids. There are two crystallographically unique Cl atoms that both are half occupied. Thermogravimetric studies show that the compound starts to decompose at 500°C. The crystal structure of the new compound is closely related to the previously described compound Cd4(SeO3)2Cl4(H2O).

Role of calcium and EPAC in norepinephrine-induced ghrelin secretion.

Ghrelin is an orexigenic hormone secreted principally from a distinct population of gastric endocrine cells. Molecular mechanisms regulating ghrelin secretion are mostly unknown. Recently, norepinephrine (NE) was shown to enhance ghrelin release by binding to ß1-adrenergic receptors on ghrelin cells. Here, we use an immortalized stomach-derived ghrelin cell line to further characterize the intracellular signaling pathways involved in NE-induced ghrelin secretion, with a focus on the roles of Ca(2+) and cAMP. Several voltage-gated Ca(2+) channel (VGCC) family members were found by quantitative PCR to be expressed by ghrelin cells. Nifedipine, a selective L-type VGCC blocker, suppressed both basal and NE-stimulated ghrelin secretion. NE induced elevation of cytosolic Ca(2+) levels both in the presence and absence of extracellular Ca(2+). Ca(2+)-sensing synaptotagmins Syt7 and Syt9 were also highly expressed in ghrelin cell lines, suggesting that they too help mediate ghrelin secretion. Raising cAMP with the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine also stimulated ghrelin secretion, although such a cAMP-mediated effect likely does not involve protein kinase A, given the absence of a modulatory response to a highly selective protein kinase A inhibitor. However, pharmacological inhibition of another target of cAMP, exchange protein-activated by cAMP (EPAC), did attenuate both basal and NE-induced ghrelin secretion, whereas an EPAC agonist enhanced basal ghrelin secretion. We conclude that constitutive ghrelin secretion is primarily regulated by Ca(2+) influx through L-type VGCCs and that NE stimulates ghrelin secretion predominantly through release of intracellular Ca(2+). Furthermore, cAMP and its downstream activation of EPAC are required for the normal ghrelin secretory response to NE.

Can zinc(II) ions be doped into the crystal structure of L-proline cadmium chloride monohydrate?

The bivalent metals Cd(II) and Zn(II) exhibit different stereochemical requirements for the set of chloride and L-proline ligands, which precludes the doping of Zn(II) ions into the crystal structure of dichloro(l-proline)cadmium(II) hydrate also referred to as L-proline cadmium chloride monohydrate (L-PCCM). Hence, the reported claim of growth of zinc doped L-PCCM crystals namely Zn(0.4 mol):LPCCM and Zn(0.2 mol):LPCCM by Vetrivel et al. (S. Vetrivel, P. Anandan, K. Kanagasabapathy, S. Bhattacharya, S. Gopinath, R. Rajasekaran, Effect of zinc chloride on the growth and characterization of l-proline cadmium chloride monohydrate semiorganic NLO single crystals, Spectrochim. Acta 110A (2013) 317-323), is untenable.

In vivo biodistribution and synergistic toxicity of silica nanoparticles and cadmium chloride in mice.

Silica nanoparticles (SiNPs) are now in daily use due to their low intrinsic toxicity. Cadmium is a ubiquitous environmental pollutant. In spite of real risk of humans' co-exposure to SiNPs and cadmium, their synergistic toxicity is still unclear. Here, we report the synergistic effects of SiNPs and CdCl2 on their biodistribution and subacute toxicity in mice. The biodistributions, histopathological changes, serum biochemical parameters and oxidative stress responses were determined after intraperitoneal injection of SiNPs and/or CdCl2 to mice. SiNPs and CdCl2 have a positive synergistic toxicity in mice. Although SiNPs were low toxic to mice, co-exposure of SiNPs and CdCl2 significantly enhanced CdCl2-induced oxidative damage in the liver as indicated by the severe liver dysfunction and histopathological abnormalities. Co-exposure to SiNPs and CdCl2 markedly increased the cadmium accumulation in the liver, which induced significant hepatic oxidative stress. In vitro binding assays indicated that serum albumin and Cd(2+) mutually enhanced the binding of each other to SiNPs via the interaction of serum albumin and Cd(2+). The uptake of serum albumin- and Cd(2+)-bound SiNPs by the macrophages significantly increased cadmium accumulation in mice. These results demonstrate that serum albumins play an important role in the positive synergistic toxicity of SiNPs and CdCl2.

Synthesis, structure and antifungal activity of thiophene-2,3-dicarboxaldehyde bis(thiosemicarbazone) and nickel(II), copper(II) and cadmium(II) complexes: unsymmetrical coordination mode of nickel complex.

The reaction of nickel(II), copper(II) chlorides and cadmium(II) chloride and bromide with thiophene-2,3-dicarboxaldehyde bis(thiosemicarbazone) (2,3BTSTCH2) leads to a series of new complexes: [Ni(2,3BTSTCH)]Cl, [Cu(2,3BTSTC)], [CdCl2(2,3BTSTCH2)] and [CdBr2(2,3BTSTCH2)]. The crystal structures of the ligand and of [Ni(2,3BTSTCH)]Cl complex have been determined. In this case, we remark an unusual non-symmetrical coordination mode for the two functional groups: one acting as a thione and the second as a deprotonated thiolate. All compounds have been tested for their antifungal activity against human pathogenic fungi: Candida albicans, Candida glabrata and Aspergillus fumigatus, the cadmium complexes exhibit the highest antifungal activity. Cytotoxicity was evaluated using two biological methods: human MRC5 cultured cells and brine shrimp Artemia salina bioassay.

Effect of zinc chloride on the growth and characterization of L-proline cadmium chloride monohydrate semiorganic NLO single crystals.

Single crystals of zinc doped L-proline cadmium chloride monohydrate were successfully grown from aqueous solution by slow evaporation method at room temperature for different molar concentration of zinc chloride. The structural properties of grown crystals have been studied by single crystal X-ray diffraction, powder X-ray diffraction studies and Fourier transform infrared spectral analysis. The incorporation of the dopant (zinc chloride) into L-proline cadmium chloride monohydrate crystal lattice has been confirmed by EDAX analysis. UV-Vis spectral analyses showed that the doped crystals have lower UV cut-off wavelength at 200 nm combined with very good transparency about 85% in a very wide range. The second harmonic generation efficiency test has been carried out and results are discussed. The 0.2 and 0.4 mol Zinc chloride doped crystals were thermally stable up to 208.9 °C and 211.9 °C respectively. The electrical properties have been studied by dielectric constant studies. All results are compared with the results of pure L-PCCM crystals.

Field-effect transistors from lithographically patterned cadmium selenide nanowire arrays.

Field-effect transistors (NWFETs) have been prepared from arrays of polycrystalline cadmium selenide (pc-CdSe) nanowires using a back gate configuration. pc-CdSe nanowires were fabricated using the lithographically patterned nanowire electrodeposition (LPNE) process on SiO(2)/Si substrates. After electrodeposition, pc-CdSe nanowires were thermally annealed at 300 °C × 4 h either with or without exposure to CdCl(2) in methanol-a grain growth promoter. The influence of CdCl(2) treatment was to increase the mean grain diameter from 10 to 80 nm as determined by grazing incidence X-ray diffraction and to convert the crystal structure from cubic to wurtzite. Measured transfer characteristics showed an increase of the field effect mobility (µ(eff)) by an order of magnitude from 1.94 × 10(-4) cm(2)/(V s) to 23.4 × 10(-4) cm(2)/(V s) for pc-CdSe nanowires subjected to the CdCl(2) treatment. The CdCl(2) treatment also reduced the threshold voltage (from 20 to 5 V) and the subthreshold slope (by ~35%). Transfer characteristics for pc-CdSe NWFETs were also influenced by the channel length, L. For CdCl(2)-treated nanowires, µ(eff) was reduced by a factor of eight as L increased from 5 to 25 µm. These channel length effects are attributed to the presence of defects including breaks and constrictions within individual pc-CdSe nanowires.

The CdCl2 effects on synthetic DNAs encaged in the nanodomains of a cationic water-in-oil microemulsion.

The present work is dedicated to the study of the interactions of CdCl(2) with the synthetic polynucleotides polyAT and polyGC confined in the nanoscopic aqueous compartment of the water-in-oil microemulsion CTAB/pentanol/hexane/water, with the goal to mimic in vitro the situation met by the nucleic acids in vivo. In biological structures, in fact, very long strings of nucleic acids are segregated into very small compartments having a radius exceedingly smaller than the length of the encapsulated macromolecule. For comparison, the behaviour of polyGC was also studied in aqueous solutions of matched composition. The conformational and thermal stabilities of both polynucleotides enclosed in the inner compartment of the microemulsion are scarcely affected by the presence of CdCl(2), whereas in solution immediate and large effects were observed also at room temperature. The lack of effects of CdCl(2) on the properties of the biopolymers entrapped in the aqueous core of the microemulsion has been attributed to the peculiar characteristics of the medium (low dielectric constant, in particular) which cause a total repression of the CdCl(2) dissociation that is not complete even in water. In fact, several of the numerous effects of CdCl(2) observed on the conformational stability of polyGC in aqueous solutions have also been ascribed to the limited dissociation of the cadmium salt.