cytochrome-c-t and cadmium-telluride

We applied transient absorption spectroscopy to study the early photodynamics in a system composed of CdTe quantum dots (QDs) and cytochrome c (Cyt c) protein. In the QDs and Cyt c mixtures, about 25 % of the excited QD electrons quickly relax (∼23 ps) to the ground state and roughly 75 % decay on slower time scale - mostly due to quenching by Cyt c. On the basis of the assumed model, we estimated the contribution of electron transfer and other mechanisms to this quenching. The primary quenching mechanism is probably energy transfer but electron transfer makes a significant contribution (∼8 %), resulting in photoreduction of Cyt c. The lifetime of one fraction of reduced Cyt c (35-90 %) is ∼ 1 ms and the lifetime of the remaining fraction was longer than the ∼ 50-ms time window of the experiment. We speculate that, in the former fraction, the back electron transfer from the reduced Cyt c to QDs occurs and the latter fraction of Cyt c is stably reduced.

Topics: Cadmium Compounds; Cytochromes c; Electrons; Quantum Dots; Tellurium

Recently, the use of CdTe quantum dots in the field of biomedicine, such as biological imaging, biosensors, cell markers, and drug carriers, is increasing due to their special physical and chemical properties. However, their biosafety assessment lags far behind their rapid application. In this study, we observed that CdTe quantum dots with certain exposed doses and time decreased the cell viability and increased the apoptosis rates in ND7/23 cells. In general, CdTe quantum dots exposure could promote the accumulation of reactive oxygen species (ROS) in cells and decrease the mitochondrial membrane potential, which led to pathological changes and subcellular organelle damages. We hypothesized that the mitochondrial pathway could be involved in CdTe quantum dots-induced apoptosis. The results suggested that CdTe quantum dots exposure increased the expression levels of three mitochondrial pathway markers, for example, caspase-3, cytochrome c, and Bax while decreased Bcl-2 protein expression, following with cytochrome c falling out of the inner membrane of mitochondrial and releasing into the cytoplasm. The application of caspase-3 protein inhibitor Ac-DEVD-CHO could decrease apoptosis rates in ND7/23 cells. The results, taken together, demonstrated that CdTe quantum dots could induce apoptosis of ND7/23 cells through the mitochondrial pathway. Our findings provide a novel insight for researchers to explore CdTe quantum dots' toxic mechanisms to reduce their adverse effects.

Topics: Apoptosis; Cadmium Compounds; Caspase 3; Cell Line; Cytochromes c; Ganglia, Spinal; Quantum Dots; Tellurium

In the current study, the cytotoxicity and mechanisms of cadmium telluride quantum dots (CdTe QDs) on RSC96 cells were evaluated by exposing different doses of CdTe QDs for 24 h. Two types of cell death, including apoptosis and autophagy, as well as two important organelles, mitochondria and endoplasmic reticulum, were focused after CdTe QDs exposure. The results showed that CdTe QDs induced apoptosis in RSC96 cells in a concentration-dependent manner; promoted the accumulation of intracellular reactive oxygen species; decreased the mitochondrial membrane potential; caused the release of cytochrome c; and also increased the expression of Bcl-2 associated X protein, caspase-3, and cytochrome c proteins and decreased the expression of Bcl-2 protein. Further results also confirmed that CdTe QDs could be internalized by RSC96 cells, and the exposure and internalization of CdTe QDs could induce excessive endoplasmic reticulum stress in the cells, and the expression levels of binding immunoglobulin protein, C/EBP homologous protein, and caspase12 proteins were increased in a concentration-dependent manner. Moreover, autophagy-related proteins LC3II, Beclin1, and P62 all increased after CdTe QDs exposure, suggesting that CdTe QDs exposure both promoted autophagosome formation and inhibited autophagosome degradation, and that CdTe QDs affected the autophagic flow in RSC96 cells. In conclusion, CdTe QDs are able to cause apoptosis and autophagy in RSC96 cells through mitochondrial and endoplasmic reticulum stress pathways, and the possible neurotoxicity of CdTe QDs should be further investigated.

Topics: Apoptosis; Autophagy; Cadmium Compounds; Cytochromes c; Endoplasmic Reticulum Stress; Oxidative Stress; Proto-Oncogene Proteins c-bcl-2; Quantum Dots; Rats; Schwann Cells; Tellurium

Utilizing fluorescence spectroscopy, non-covalent and covalent interactions of L-cys@CdTe quantum dots to bovine serum albumin (BSA), cytochrome c and trypsin were investigated. L-cys@CdTe QDs with the emission maximum at 530 nm and an average diameter of 2.6 nm were synthesized in the aqueous medium. Formaldehyde, N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (EDC) with N-hydroxysuccinimide (NHS), and glutaraldehyde was applied as cross-linkers. In the case of both electrostatic and covalent strategies PARAFAC, as a powerful multi-way chemometrics technique, was utilized to analyze fluorescence excitation-emission (EEM) spectra. For non-covalent and covalent bonding, two and three significant components composed the PARAFAC models. Resolved EEM shows that in the presence of formaldehyde, a new component with an emission peak similar to BSA was obtained. Using EDC-NHS cross-linker, the fluorescence peak of the newly formed component was in a distinct wavelength with similar emission intensity, compared to L-cys@CdTe QDs and BSA. Employing glutaraldehyde, a distinguished component was easily detected at emission wavelengths higher than that of L-cys@CdTe QDs and proteins. It was concluded that the choice of cross-linker is a critical step to create different emission spectra when dealing with nano-bio-conjugations. This study shows that glutaraldehyde cross-linker leads to increase sensitivity, selectivity, and accuracy of protein analysis.

Topics: Cadmium Compounds; Cytochromes c; Quantum Dots; Serum Albumin, Bovine; Spectrometry, Fluorescence; Static Electricity; Tellurium; Trypsin

Deoxynivalenol (DON) is considered a mycotoxin that is toxic to the agricultural environment and human body. It is necessary to study the pathophysiological mechanism of DON toxicity at the cellular level. Cytochrome c (Cyt c), as an important biomarker of DON-induced apoptosis that may lead to a bipartite 'point-of-no return' event, is of great significance to be detected using cell imaging. Herein, we synthesized a DON-deactivated emission fluorescent probe, the molecularly imprinted polymer-coated quantum dots (CdTe@MIP), for monitoring the Cyt c level with a photoinduced electron transfer strategy. The CdTe@MIP probe can be easily loaded into cells and perform well due to its great sensitivity and selectivity and its fluorescence was gradually quenched with the increasing concentration (0-10 μM) and incubation time (0-7.5 h) of DON. Cell imaging results of apoptosis induced by DON was consistent with that of the cell counting kit-8 assay and flow cytometry technique. The developed method can be used to monitor DON-induced apoptosis and provide an early-warning system for the contaminant toxicity.

Topics: Apoptosis; Cadmium Compounds; Cytochromes c; Electrons; Fluorescent Dyes; Humans; Quantum Dots; Tellurium; Trichothecenes

Cytochrome c (Cyt c) is commonly used as intrinsic biomarker for several characteristics of the cell such as respiration, energy level and apoptosis. In the present study a simple colorimetric sensor should be developed and tested for the real-time detection of Cyt c in living cells. We synthesized cadmium telluride quantum dots (CdTe QDs) capped with thioglycolic acid (TGA) as a fluorometric Cyt c nanosensor. The synthesized TGA/CdTe QDs nanosensor was characterized by Fourier transform infrared spectroscopy, transmission electron microscopy, and absorption as well as fluorescence spectrophotometry. We investigated the developed TGA/CdTe QDs sensor with regard to its applicability in the fluorometric detection of Cyt c. Results showed that the TGA/CdTe QDs could be used as a sensitive fluorescence probe for the quantification of different concentrations of Cyt c ranging from 0.5 - 2.5μM. Increased binding of QDs to Cyt c results in decreasing fluorescence. The fluorescence of the QDs is inversely correlated to the Cyt c concentration. Based on these data, a standard curve up to 2.5μM Cyt c was established. Moreover, the developed nanosensor was applied in different concentrations on primary human dermal fibroblasts. Results showed that TGA/CdTe QDs were taken up by cells and could be visualized by fluorescence microscopy. Quantification of Cyt c within living cells via QDs is, however, influenced by various factors such as cell damage, QD aggregation or the level of reactive oxygen species, which have to be taken into account.

Topics: Biosensing Techniques; Cadmium Compounds; Cytochromes c; Fluorescent Dyes; Humans; Microscopy, Electron, Transmission; Quantum Dots; Spectrometry, Fluorescence; Tellurium

Monitoring of bio-catalytic events by using nano-probes is of immense interest due to unique optical properties of metal nanoparticles. In the present study, tunneling of enzyme activity was achieved using redox cofactors namely oxidized cytochrome-c (Cyt-c) and Co-enzyme-Q (Co-Q) immobilized on Quantum dots (QDs) which acted as a bio-probe for NAD(+) dependent dehydrogenase catalyzed reaction. We studied how electron transfer from substrate to non-native electron acceptors can differentially modify photoluminescence properties of CdTe QDs. Two probes were designed, QD-Ox-Cyt-c and QD-Ox-Co-Q, which were found to quench the fluorescence of QDs. However, formaldehyde dehydrogenase (FDH) catalyzed reduction of Cyt-c and Co-Q on the surface of QDs lead to fluorescence turn-on of CdTe QDs. This phenomenon was successfully used for the detection of HCHO in the range of 0.01-100,000ng/mL (LOD of 0.01ng/mL) using both QD-Ox-Cyt-c (R(2)=0.93) and QD-Ox-Co-Q (R(2)=0.96). Further probe performance and stability in samples like milk, wine and fruit juice matrix were studied and we could detect HCHO in range of 0.001-100,000ng/mL (LOD of 0.001ng/mL) with good stability and sensitivity of probe in real samples (R(2)=0.97). Appreciable recovery and detection sensitivity in the presence of metal ions suggests that the developed nano-probes can be used successfully for monitoring dehydrogenase based bio-catalytic events even in the absence of NAD(+). Proposed method is advantageous over classical methods as clean up/ derivatization of samples is not required for formaldehyde detection.

Topics: Aldehyde Oxidoreductases; Animals; Biosensing Techniques; Cadmium Compounds; Cytochromes c; Electron Transport; Enzymes, Immobilized; Food Analysis; Formaldehyde; Fruit and Vegetable Juices; Milk; NAD; Oxidation-Reduction; Quantum Dots; Tellurium; Ubiquinone; Wine

Self-assembly of proteins and inorganic nanoparticles into terminal assemblies makes possible a large family of uniformly sized hybrid colloids. These particles can be compared in terms of utility, versatility and multifunctionality to other known types of terminal assemblies. They are simple to make and offer theoretical tools for designing their structure and function. To demonstrate such assemblies, we combine cadmium telluride nanoparticles with cytochrome C protein and observe spontaneous formation of spherical supraparticles with a narrow size distribution. Such self-limiting behaviour originates from the competition between electrostatic repulsion and non-covalent attractive interactions. Experimental variation of supraparticle diameters for several assembly conditions matches predictions obtained in simulations. Similar to micelles, supraparticles can incorporate other biological components as exemplified by incorporation of nitrate reductase. Tight packing of nanoscale components enables effective charge and exciton transport in supraparticles and bionic combination of properties as demonstrated by enzymatic nitrate reduction initiated by light absorption in the nanoparticle.

Topics: Cadmium Compounds; Colloids; Cytochromes c; Humans; Metal Nanoparticles; Molecular Docking Simulation; Nitrate Reductase; Pichia; Static Electricity; Tellurium

The mechanisms of toxicity related to human hepatocellular carcinoma HepG2 cell exposures to cadmium telluride quantum dots (CdTe-QDs) were investigated. CdTe-QDs caused cytotoxicity in HepG2 cells in a dose- and time-dependent manner. Treated cells showed an increase in reactive oxygen species (ROS). Altered antioxidant levels were demonstrated by depletion of reduced glutathione (GSH), a decreased ratio of reduced glutathione to oxidized glutathione (GSH/GSSG) and an increased NF-E2-related Factor 2 (Nrf2) activation. Enzyme assays showed that superoxide dismutase (SOD) activity was elevated whereas catalase (CAT) and glutathione-S-transferase (GST) activities were depressed. Further analyses revealed that CdTe-QD exposure resulted in apoptosis, indicated by changes in levels of caspase-3 activity, poly ADP-ribose polymerase (PARP) cleavage and phosphatidylserine externalization. Extrinsic apoptotic pathway markers such as Fas levels and caspase-8 activity increased as a result of CdTe-QD exposure. Involvement of the intrinsic/mitochondrial apoptotic pathway was indicated by decreased levels of B-cell lymphoma 2 (Bcl2) protein and mitochondrial cytochrome c, and by increased levels of mitochondrial Bcl-2-associated X protein (Bax) and cytosolic cytochrome c. Further, mitogen-activated protein kinases (MAPKs) such as c-Jun N-terminal kinases (JNK), extracellular signal-regulated kinases (Erk1/2), and p38 were all activated. Our findings reveal that CdTe-QDs cause oxidative stress, interfere with antioxidant defenses and activate protein kinases, leading to apoptosis via both extrinsic and intrinsic pathways. Since the effects of CdTe-QDs on selected biomarkers were similar or greater compared to those of CdCl2 at equivalent concentrations of cadmium, the study suggests that the toxicity of CdTe-QDs arises from a combination of the effects of cadmium and ROS generated from the NPs.

Topics: Apoptosis; Cadmium Compounds; Carcinoma, Hepatocellular; Catalase; Cell Survival; Cytochromes c; Glutathione; Glutathione Disulfide; Hep G2 Cells; Humans; Liver Neoplasms; Microscopy, Confocal; Mitogen-Activated Protein Kinases; NF-E2-Related Factor 2; Oxidative Stress; Quantum Dots; Signal Transduction; Superoxide Dismutase; Tellurium

A newly designed molecularly imprinted polymer (MIP) material was fabricated and successfully utilized as recognition element to develop a quantum dots (QDs) based MIP-coated composite for selective recognition of the template cytochrome c (Cyt). The composites were synthesized by sol-gel reaction (imprinting process). The imprinting process resulted in an increased affinity of the composites toward the corresponding template. The fluorescence of MIP-coated QDs was stronger quenched by the template versus that of non-imprinted polymer (NIP)-coated QDs, which indicated the composites could recognize the corresponding template. The results of specific experiments further exhibited the recognition ability of the composites. Under optimum conditions, the linear range for Cyt is from 0.97 μM to 24 μM, and the detection limit is 0.41 μM. The new composites integrated the high selectivity of molecular imprinting technology and fluorescence property of QDs and could convert the specific interactions between imprinted cavities and corresponding template to the obvious changes of fluorescence signal. Therefore, a simple and selective sensing system for protein recognition has been realized.

Topics: Biosensing Techniques; Cadmium Compounds; Cytochromes c; Equipment Design; Equipment Failure Analysis; Molecular Probe Techniques; Polymers; Quantum Dots; Spectrometry, Fluorescence; Surface Properties; Tellurium

Quantum dots (QDs) are luminescent nanoparticles with unique optical properties that have been exploited for single-cell and whole-animal imaging. When coated with proteins or biocompatible polymers, QDs are not deleterious to cells and organisms. However, when QDs are retained in cells or accumulated in the body for a long period of time, their coatings may be degraded, yielding "naked" QDs. Here, we show that "naked" QDs induce damage to the plasma membrane, mitochondrion, and nucleus, leading to cell death. Reactive oxygen species (ROS) are important players in mediating QD-induced cellular damage. QD-induced cytotoxicity can be reduced or even eliminated without covalent binding of protective agents to the QD surface. Results from these studies suggest the critical role of several subcellular compartments in QD-induced cytotoxicity and point toward multiple molecular targets in nonclassical apoptosis.

Topics: Acetylcysteine; Cadmium Compounds; Caspases; Cell Death; Cell Line, Tumor; Cell Membrane; Cell Nucleus; Cytochromes c; Humans; Mitochondria; Organelles; Quantum Dots; Reactive Oxygen Species; Tellurium