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Article

Cis-[Cr(C2O4)(pm)(OH2)2]+ Coordination Ion as a Specific Sensing Ion for H2O2 Detection in HT22 Cells

1
Faculty of Chemistry, University of Gdańsk, Wita Stwosza 63, Gdańsk 80-308, Poland
2
Department of Histology, Medical University of Gdańsk, Dębinki 1, Gdańsk 80-211, Poland
3
Department of Medical Chemistry, Medical University of Gdańsk, Dębinki 1, Gdańsk 80-211, Poland
*
Author to whom correspondence should be addressed.
Molecules 2014, 19(6), 8533-8543; https://doi.org/10.3390/molecules19068533
Submission received: 8 April 2014 / Revised: 30 May 2014 / Accepted: 10 June 2014 / Published: 23 June 2014
(This article belongs to the Section Medicinal Chemistry)

Abstract

:
The purpose of this study was to examine the application of the coordinated cis-[Cr(C2O4)(pm)(OH)2]+ cation where pm denotes pyridoxamine, as a specific sensing ion for the detection of hydrogen peroxide (H2O2). The proposed method for H2O2 detection includes two key steps. The first step is based on the nonenzymatic decarboxylation of pyruvate upon reaction with H2O2, while the second step is based on the interaction of cis-[Cr(C2O4)(pm)(OH2)2]+ with the CO2 released in the previous step. Using this method H2O2 generated during glutamate-induced oxidative stress was detected in HT22 hippocampal cells. The coordination ion cis-[Cr(C2O4)(pm)(OH2)2]+ and the spectrophotometric stopped-flow technique were applied to determine the CO2 concentration in cell lysates, supernatants and cell-free culture medium. Prior to CO2 assessment pyruvate was added to all samples studied. Pyruvate reacts with H2O2 with 1:1 stoichiometry, and consequently the amount of CO2 released in this reaction is equivalent to the amount of H2O2.

Graphical Abstract

1. Introduction

Reactive oxygen species (ROS) generation contributes to the ethiology of many diseases, including diabetes, arteriosclerosis or neurodegenerative disorders and others [1,2,3]. The increase in intracellular production of free radicals may lead to cellular damage, including alterations of lipids, proteins and DNA [4]. ROS [5,6], in particular superoxide anion (O2•−), hydroxyl radical (OH) or hydrogen peroxide (H2O2) and reactive nitrogen species [7,8,9,10], e.g., nitrogen dioxide, are highly cytotoxic. Superoxide anion is able to react with nitric oxide to form toxic peroxinitrite anions or dismutate into H2O2 which in turn can be transformed into highly reactive and toxic hydroxyl radicals. H2O2 is one of the most important mediators of oxidative stress detected under pathological conditions. It was observed that under pathological conditions such as ischemia-reperfusion injury, excessive production of H2O2 occurs. H2O2 is probably involved in the neuronal damage seen in Parkinson’s and Huntington’s diseases and other neuronal disorders.
Oxidative stress can be evaluated by detecting ROS using biosensors [11]. However, in most cases, the concentration of ROS is evaluated without enough precision [12,13,14]. The other limitations include not enough sensitivity or lack of specificity. Therefore, there is a need for developing more specific, sensitive and effective methods for the detection and measurement of intracellular ROS levels [12]. Pyruvic acid, like other α-ketoacids, acts as a H2O2 scavenger and is able to react non-enzymatically with H2O2 yielding the following products: acetic acid, H2O and CO2. Interestingly, pyruvate is present in mammalian cells and its antioxidative properties contribute to the cellular defense against H2O2-mediated cytotoxicity [15]. In the present study, a new method of the H2O2 concentration assessment has been demonstrated. This method is based on the ability of the molecular biosensor—coordinate ion cis-[Cr(C2O4)(pm)(OH)2]+ to effectively trap CO2, one of the final products of the chemical reaction between H2O2 and exogenous pyruvate (1). Noteworthily, the amount of CO2 released in this reaction is equivalent to the amount of H2O2:
pyruvate + H2O2 → acetate + CO2 + H2O
Glutamate is a neurotransmitter in the central nervous system. It was reported to induce neuronal cell death at mM levels of concentrations [16]. High concentrations of extracellular glutamate inhibit the glutamate/cystine antiporter, which results in the depletion of intracellular glutathione that converts H2O2 to H2O [17]. Typically, 5 mM glutamate was shown to induce oxidative stress in HT22 cells leading to death [18]. Therefore, as an experimental model to study the generation of H2O2, mouse hippocampal HT22 cells treated with 5 mM l-glutamate were used in our study.
Previously, our research group reported a quantitative determination of H2O2 in osteosarcoma cells [19]. In this method a molecular biosensor was used, the most effective being a coordination complex ion cis-[Cr(C2O4)(pm)(OH2)2]+. The CO2 uptake was studied using a spectrophotometric stopped-flow method whereby it was possible to determine the content of H2O2 in the biological material under anaerobic conditions. The method described is based on the assumption of the selective reaction of α-keto acid—pyruvate with H2O2, with the subsequent decarboxylation of the intermediate product, pyruvic peracid, and the capture of the CO2 released. The results provided arguments for the usefulness of pyruvate application for cell culture studies where culture media could produce significant levels of H2O2 before treatment of cells. On the other hand endogenous and exogenous sources of H2O2 implicated in cytotoxity in a variety of human diseases can be safely prevented by pyruvate. The efficiency of this scavenger was clearly demonstrated by a novel application of a molecular CO2 detection method based on cis-[Cr(C2O4)(pm)(OH2)2]+ ion.

2. Results and Discussion

The cis-[Cr(C2O4)(pm)(OH2)2]+ ion was previously developed as a specific molecular biosensor to detect uptake of CO2, generated in the reaction between Na2CO3 and HCl [20]. To do this the reaction between the cis-[Cr(C2O4)(pm)(OH2)2]+ ion and carbon dioxide in aqueous solution was monitored between 340 nm and 700 nm using a spectrophotometric stopped-flow method. It was observed that the reaction of carbon dioxide uptake by the applied biosensor ran in two-steps. The first step was about 50 times faster than the second one. This conclusion is based on the analytical results. The analytical model was already described in [21]. During the carbon dioxide uptake (where CO2 was generated in a chemical reaction) by cis-[Cr(C2O4)(pm)(OH2)2]+ complex ion the most significant changes of absorbance were seen at λ = 560 nm. Consequently, on the basis of results obtained a two-step mechanism for the uptake of carbon dioxide by cis-[Cr(C2O4)(pm)(OH2)2]+ ion was proposed and described [21].
Having the above knowledge concerning a chemical model, the same coordination compound of Cr(III) with a bidendate ligand—pyridoxamine—was checked and successfully applied in this study also in biological material, namely for the detection of CO2 generated during the glutamate-induced oxidative stress in HT22 cells. The results obtained are presented in Figure 1a For comparison, the previously obtained results of global analysis (GA) for reaction of CO2 uptake by the cis-[Cr(C2O4)(pm)(OH2)2]+ ion in a chemical model within the consecutive reaction model (A→B→C) are presented in Figure 1b. In Figure 1a,b symbol “A” means the substrate cis-[Cr(C2O4)(pm)(OH2)2]+, “B”- intermediate product, and “C”- the final product cis-[Cr(C2O4)(pm)(O2CO)]. The different behaviour of A, B and C is the result of decrease in the substrate concentration and product formation. As seen the same absorption maxima can be observed for both systems – chemical and biological. This conformity can be treated as a confirmation that the proposed chemical model fits the biological system. Figure 1a shows the global analysis results for the reaction of CO2 uptake from biological material by the cis-[Cr(C2O4)(pm)(OH2)2]+ ion. Cr(III) is inert and this causes the reaction to be slowed down. To confirm the mechanism of uptake previously proposed on the basis of the chemical reaction system [21] in the biological model, in the first step (carbon dioxide uptake), kinetic data were fitted by a simple A→B reaction model (where B denotes the intermediate). Furthermore, in the second step (the closure of the ring of carbonate ion) [22], the reaction was monitored at the wavelength where the maximum difference in molar absorptivities between the intermediate products and products (B→C reaction model) was observed, at λ = 560 nm (Figure 1). It should be pointed that the results obtained by the global analysis (GA) [23] method were confirmed by another independent method of the singular value decomposition (SVD) [22] analysis (Figure 1). GA and SVD are the mathematical methods used in calculations.
Figure 1. Comparison of the biological and chemical reaction models. (a) The biological reaction model (I) Curves of concentration decay and buildup of the substrate A (which is the cis-[Cr(C2O4)(pm)(OH2)2]+ ion), product C as cis-[Cr(C2O4)(pm)(O2CO)] ion, and intermediate product B. (II) Absorption spectra of the reactants A, B and C - the detection of CO2 generated during glutamate induced oxidative stress in HT22 cells. (b) The chemical reaction model (I) Curves of concentration decay and buildup of the substrate A (which is the cis-[Cr(C2O4)(pm)(OH2)2]+ ion), product C as cis-[Cr(C2O4)(pm)(O2CO)] ion, and intermediate product B. (II) Absorption spectra of the reactants A, B and C at pH = 7.13, t = 20 °C.
Figure 1. Comparison of the biological and chemical reaction models. (a) The biological reaction model (I) Curves of concentration decay and buildup of the substrate A (which is the cis-[Cr(C2O4)(pm)(OH2)2]+ ion), product C as cis-[Cr(C2O4)(pm)(O2CO)] ion, and intermediate product B. (II) Absorption spectra of the reactants A, B and C - the detection of CO2 generated during glutamate induced oxidative stress in HT22 cells. (b) The chemical reaction model (I) Curves of concentration decay and buildup of the substrate A (which is the cis-[Cr(C2O4)(pm)(OH2)2]+ ion), product C as cis-[Cr(C2O4)(pm)(O2CO)] ion, and intermediate product B. (II) Absorption spectra of the reactants A, B and C at pH = 7.13, t = 20 °C.
Molecules 19 08533 g001
In this study a new method for H2O2 detection in cell lysates, supernatants and cell-free culture medium is demonstrated. This method is based on both the interaction of the coordination compound cis-[Cr(C2O4)(pm)(OH)2]+ with CO2 and the nonenzymatic reaction of pyruvate with H2O2. Using cis-[Cr(C2O4)(pm)(OH)2]+ and the spectrophotometric stopped-flow technique the CO2 concentration in cell lysates, supernatants and culture medium was determined. Since pyruvate reacts nonenzymatically with H2O2 with 1:1 stoichiometry releasing CO2, the amount of H2O2 in this reaction is equivalent to the amount of CO2 released. Therefore, prior to CO2 assessment pyruvate was added to all samples studied.
As an experimental model to study the generation of H2O2, mouse hippocampal HT22 cells treated with sodium 5 mM l-glutamate were used. Five mM glutamate was used to induce the oxidative stress in HT22 cells, which will subsequently lead to cell death [18]. In agreement with this data, our results showed that the viability of HT22 cells treated with 5 mM l-glutamate for 24 h decreased dramatically (Figure 2).
Figure 2. Antiproliferative effect of l-glutamate. HT22 cells were treated with 5 mM l-glutamate for 24 h. The cell viability was assessed using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. Data are presented as mean ± SD. *** p <0.001, statistically significant differences compared to control (untreated) cells.
Figure 2. Antiproliferative effect of l-glutamate. HT22 cells were treated with 5 mM l-glutamate for 24 h. The cell viability was assessed using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. Data are presented as mean ± SD. *** p <0.001, statistically significant differences compared to control (untreated) cells.
Molecules 19 08533 g002
In the experiments, the main source of CO2 measured in cell lysates was the reaction of pyruvate with H2O2 produced endogenously in HT22 cells treated with 5 mM l-glutamate for 24 h. In order to determine the most effective concentration of sodium pyruvate required to scavenge H2O2 present in cell lysates, the following concentrations of sodium pyruvate were tested: 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10 mM. The CO2 levels in lysates of control (untreated) cells and lysates of cells treated with 5 mM l-glutamate, measured without the addition of pyruvate, were 1.09 μM (± 0.06) and 1.1 μM (±0.05), respectively. It has been found that upon addition of 0.5–4 mM sodium pyruvate to the lysates of l-glutamate-treated HT22 cells, the CO2 level gradually increased (Figure 3).
Noteworthily, at pyruvate concentrations ranging from 5 mM to 10 mM only slight changes in the CO2 level measured in the cell lysates were observed—the CO2 level did not increase significantly. These results suggest that the 5 mM–10 mM concentration range of sodium pyruvate can be used in the proposed method to effectively assess the CO2 concentration in the cell lysates obtained upon lysis of l-glutamate-treated HT22 cells.
H2O2 has the ability to penetrate biological membranes, which enables it to be released from the cells where it is produced and thus affect neighbouring cells [5]. Therefore, the CO2level in supernatants—the surrounding of HT22 cells was assessed. Noteworthily, the source of CO2 detected in supernatants can also be H2O2 production resulting from oxidation of components of culture medium [24]. The results revealed that the CO2 levels in supernatants separated from control (untreated) cells and those collected after separation of cells treated with 5 mM l-glutamate, both measured without the addition of pyruvate, were 0.94 μM (± 0.05) and 1.03 μM (± 0.22), respectively. Moreover, it has been found that upon addition of 0.5–6 mM sodium pyruvate to the supernatants separated from l-glutamate-treated HT22 cells, the CO2 level gradually increased (Figure 3). At pyruvate concentrations ranging from 7 mM to 10 mM only slight changes in the CO2 level in the supernatants were observed—the CO2 level did not increased significantly. These results indicate that among the pyruvate concentrations tested, the concentration range 7-10 mM is sufficient to assess the CO2 level in the supernatants collected upon separation of HT22 cells treated with 5 mM l-glutamate.
Figure 3. CO2 assessment in cell lysates and supernatants upon treatment with 5 mM sodium l-glutamate. HT22 cells were incubated with 5 mM sodium l-glutamate for 24 h. After treatment, cells and superntants were separated. The cells were then lysed using a lysis buffer. Prior to CO2 measurement sodium pyruvate was added (final concentations of sodium pyruvate: 0.5–10 mM, respectively) to the samples containing cell lysates and supernatants, respectively. The CO2 level was assessed using a stopped-flow technique. Data are expressed as mean ± SD of three independent experiments. * p < 0.05, ** p < 0.01, statistically significant differences compared to the sample treated with 5 mM l-glutamate alone, without addition of sodium pyruvate; control - untreated cells.
Figure 3. CO2 assessment in cell lysates and supernatants upon treatment with 5 mM sodium l-glutamate. HT22 cells were incubated with 5 mM sodium l-glutamate for 24 h. After treatment, cells and superntants were separated. The cells were then lysed using a lysis buffer. Prior to CO2 measurement sodium pyruvate was added (final concentations of sodium pyruvate: 0.5–10 mM, respectively) to the samples containing cell lysates and supernatants, respectively. The CO2 level was assessed using a stopped-flow technique. Data are expressed as mean ± SD of three independent experiments. * p < 0.05, ** p < 0.01, statistically significant differences compared to the sample treated with 5 mM l-glutamate alone, without addition of sodium pyruvate; control - untreated cells.
Molecules 19 08533 g003
As mentioned previously, the culture medium may be a source of H2O2 generation [24]. Moreover, it is important to evaluate whether the CO2 level assessed by our method can be influenced by the HCO3/CO2 buffer system used in growth media to maintain the proper pH. Therefore, in addition to the CO2 assessment in cell lysates and supernatants, the CO2 level in cell-free complete culture medium was examined, under conditions of the experiment not resulting from endogenous (cellular) production of H2O2. The CO2 concentration was measured without addition or upon addition of different concentrations of sodium pyruvate, respectively. The culture medium were treated with 5 mM l-glutamate for 24 h and then collected. The CO2 concentration in control (untreated) medium sample and those treated with 5 mM l-glutamate, both measured without the addition of pyruvate, were 1.06 μM (± 0.04) and 1.09 μM (± 0.02), respectively. It has been found that upon addition of 0.5–10 mM sodium pyruvate, the CO2 level gradually increased (Figure 4). As mentioned previously, one possible explanation is that the culture medium may also be a source of H2O2 generation [19].
Figure 4. CO2 assessment in cell-free complete culture medium upon treatment with 5 mM sodium l-glutamate. The cell-free complete medium was incubated with 5 mM sodium l-glutamate for 24 h. After treatment, sodium pyruvate was added (final concentations of sodium pyruvate: 0.5–10 mM, respectively). The CO2 level was assessed using a stopped-flow technique. Data are expressed as mean ± SD. * p < 0.05, statistically significant differences compared to the sample treated with 5 mM l-glutamate alone, without addition of sodium pyruvate; control – untreated sample.
Figure 4. CO2 assessment in cell-free complete culture medium upon treatment with 5 mM sodium l-glutamate. The cell-free complete medium was incubated with 5 mM sodium l-glutamate for 24 h. After treatment, sodium pyruvate was added (final concentations of sodium pyruvate: 0.5–10 mM, respectively). The CO2 level was assessed using a stopped-flow technique. Data are expressed as mean ± SD. * p < 0.05, statistically significant differences compared to the sample treated with 5 mM l-glutamate alone, without addition of sodium pyruvate; control – untreated sample.
Molecules 19 08533 g004

3. Experimental Section

3.1. Chemicals

l-Glutamic acid monosodium salt monohydrate was purchased from Sigma (St. Louis, MO, USA). l-Glutamic acid monosodium salt solutions were prepared prior to use in sterile physiological saline solution. Sterile sodium pyruvate solution (100 mM) was obtained from Sigma and diluted in sterile water to desired concentrations before use. Dihydrochloride pyridoxamine was purchased from Sigma.

3.2. Reagents

The cis-[Cr(C2O4)(pm)(OH2)2]+ ion was prepared according to standard literature procedures [19]. The final products, cis-[Cr(C2O4)(L-L)(O2CO)] (where L-L denotes bidentate ligand—pyridoxamine (pm)) was synthesised by the previously reported method [21]. The complex cis-[Cr(C2O4)(pm)(OH2)2]+ was synthesised from K[Cr(C2O4)(H2O)2]·3H2O and pyridoxamine.

3.3. Cell Culture

The hippocampal neuronal HT22 cell line was kindly provided by Professor T. Grune (Institute of Biological Chemistry and Nutrition, University Hohenheim, Stuttgart, Germany). HT22 cells were maintained at 37 °C in a humidified atmosphere containing 5% (v/v) CO2 in Dulbecco’s Modified Eagle’s Medium without sodium pyruvate (Sigma), supplemented with 10% (v/v) heat-inactivated fetal bovine serum (Sigma), 100 IU/mL penicillin (Sigma) and 100 μg/mL streptomycin (Sigma).

3.4. MTT Assay

The 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) cytotoxicity assay is based on the ability of mitochondrial succinate dehydrogenase of viable cells to reduce the MTT tetrazolium salt HT22 cells were seeded in 96-well plates (8 × 103 cells per well). After a 24-hour incubation of cells with 5 mM l-glutamate, MTT (final concentration = 0.5 mg/mL) was added and the cells were incubated at 37 °C for the next 4 h. Supernatants were then removed and dimethyl sulfoxide (DMSO) was added to dissolve MTT formazan crystals. The absorbance was recorded using a microplate reader (ELx800; BioTek Instruments, Inc., Seattle, WA, USA). The viability of cells treated with 5 mM l-glutamate was expressed as the percentage of the viability of control cells (untreated with 5 mM l-glutamate).

3.5. CO2 Measurement

HT22 cells were incubated in 5 mM sodium l-glutamate for 24 h. After treatment, the cells and supernatants were separated. Cells were then washed with a phosphate buffered saline (PBS) and suspended in a lysis buffer (0.15 M NaCl, 0.005 M EDTA, 1% Triton X-100, 0.01 M Tris-HCl). Next, sodium pyruvate was added (final concentration: 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10 mM) to the samples containing cell lysates and supernatants, respectively, prior to CO2 measurement. The CO2 level in each sample was assessed using a spectrophotometric stopped-flow technique. The cell-free complete culture medium was treated with 5 mM l-glutamate for 24 h and then collected. Next, a different concentration of sodium pyruvate (final concentration: 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10 mM) was added prior to CO2 measurement. The concentrations of CO2 and H2O2 were determined using the complex cis-[Cr(C2O4)(pm)(OH2)2]+ as the biosensor.

3.6. Instrumentation

The UV-visible spectroscopy studies were conducted using a Perkin-Elmer Lambda 18 Instrument with the scan accuracy of 1 nm and 1 nm slit width at a scanning rate of 120.00 nm min−1. Kinetic measurements were carried out using a stopped-flow technique and an Applied Photophysics SX-17MV spectrophotometer. The observable rate constants and concentrations of CO2 were computed based on the global analysis using a “Glint” program [25,26,27,28,29].

3.7. Statistical Analysis

Statistical analysis was performed using Statistica 9 software (StatSoft, Kraków, Poland). Data are expressed as mean considering SD (standard deviation). Statistical differences were evaluated using the Mann-Whitney U test. Differences were considered significant at p < 0.05, p < 0.01, p < 0.001.

4. Conclusions

In this paper an application of a new method for H2O2 detection in biological samples i.e., hippocampal HT-22 cells, has been described. Moreover, the usefulness of this method, which is based on the interaction of CO2 with the coordination cation cis-[Cr(C2O4)(pm)(OH2)2]+ as a specific molecular biosensor, for the H2O2 detection in biological materials was discussed. The presented method turned out to be also handy tool to analyze the scavenging reaction of H2O2 by sodium pyruvate. Consequently, the efficiency of this scavenger was clearly demonstrated by the novel application of the H2O2 molecular biosensor—cis-[Cr(C2O4)(pm)(OH2)2]+ complex cation.

Acknowledgments

This work was financially supported by Polish Ministry of Science and Higher Education under grants N N204 132040.

Author Contributions

Dagmara Jacewicz, Joanna Pranczk, Dariusz Wyrzykowski and Lech Chmurzyński carried out the chemical part of the studies: synthesis of coordination compound and measurements of CO2 using spectrophotometric stopped-flow technique. Michał Woźniak and Kamila Siedlecka-Kroplewska carried out the MTT Assay. All authors read and approved the final manuscript.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Friedman, J.; Peleg, E.; Kagan, T.; Shnizer, S.; Rosenthal, T. Oxidative stress in hypertensive, diabetic, and diabetic hypertensive rats. Am. J. Hypertens. 2003, 16, 1049–1052. [Google Scholar]
  2. Reynolds, A.; Laurie, C.; Mosley, R.L.; Gendelman, H.E. Oxidative stress and the pathogenesis of neurodegenerative disorders. Int. Rev. Neurobiol. 2007, 82, 297–325. [Google Scholar]
  3. Schleicher, E.; Friess, U. Oxidative stress, AGE, and atherosclerosis. Kid. Int. Suppl. 2007, 106, S17–S26. [Google Scholar] [CrossRef]
  4. Halliwell, B. Antioxidants in human health and disease. Ann. Rev. Nutr. 1996, 16, 33–50. [Google Scholar] [CrossRef]
  5. Tuccio, B.; Zeghdaoui, A.; Finet, J.P.; Cerri, V.; Tordo, P. Use of new beta-phosphorylated nitrones for the spin trapping of free radicals. Res. Chem. Intermediat. 1996, 22, 393–404. [Google Scholar] [CrossRef]
  6. Alaghmand, M.; Blough, N.V. Source-dependent variation in hydroxyl radical production by airborne particulate matter. Environ. Sci. Technol. 2007, 41, 2364–2370. [Google Scholar] [CrossRef]
  7. Dąbrowska, A.; Jacewicz, D.; Łapińska, A.; Banecki, B.; Figarski, A.; Szkatuła, M.; Lehman, J.; Krajewski, J.; Kubasik-Juraniec, J.; Woźniak, M.; et al. Pivotal participation of nitrogen dioxide in l-arginine induced acute necrotizing pancreatitis: Protective role of superoxide scavenger 4-OH-TEMPO. Biochem. Biophys. Res. Commun. 2005, 326, 313–320. [Google Scholar] [CrossRef]
  8. Jacewicz, D.; Dąbrowska, A.; Wyrzykowski, D.; Pranczk, J.; Wozniak, M.; Kubasik-Juraniec, J.; Knap, N.; Siedlecka, K.; Neuwelt, A.J.; Chmurzynski, L. A Novel Biosensor for Evaluation of Apoptotic or Necrotic Effects of Nitrogen Dioxide during Acute Pancreatitis in Rat. Sensors 2010, 10, 280–291. [Google Scholar]
  9. Jacewicz, D.; Łapinska, A.; Dąbrowska, A.; Figarski, A.; Woźniak, M.; Chmurzyński, L. Reactions of NO2 with chromium(III) complexes with histamine and pyridoxamine ligands studied by the stopped-flow technique. Anal. Biochem. 2006, 350, 256–262. [Google Scholar] [CrossRef]
  10. Dąbrowska, A.; Jacewicz, D.; Chylewska, A.; Szkatula, M.; Knap, N.; Kubasik-Juraniec, J.; Wozniak, M.; Chmurzynski, L. Nitric dioxide as biologically important radical and its role in molecular mechanism of pancreatic inflammation. Curr. Pharm. Anal. 2008, 4, 183–196. [Google Scholar] [CrossRef]
  11. Nieradka, K.; Gotszalk, T.P.; Schroeder, G.M. A novel method for simultaneous readout of static bending and multimode resonance-frequency of microcantilever-based biochemical sensors. Proc. Eng. 2010, 5, 910–913. [Google Scholar] [CrossRef]
  12. Jacewicz, D.; Żamojć, K.; Wyrzykowski, D.; Chmurzyński, L. Analytical methods for determination of CO and CO2 and their applicability in biological studies. Curr. Pharm. Anal. 2013, 9, 226–235. [Google Scholar] [CrossRef]
  13. Nordin, H.; Jungnelius, M.; Karlsson, R.; Karlsson, O.P. Kinetic studies of small molecule interactions with protein kinases using biosensor technology. Anal. Biochem. 2005, 340, 359–368. [Google Scholar] [CrossRef]
  14. Schellera, F.W.; Wollenbergera, U.; Warsinkea, A.; Lisdata, F. Research and development in biosensors. Curr. Opin. Biotech. 2001, 12, 35–40. [Google Scholar] [CrossRef]
  15. O’Donnell-Tormey, J.; Nathan, C.F.; Lanks, K.; DeBoer, C.J.; de la Harpe, J. Secretion of pyruvate. An antioxidant defense of mammalian cells. J. Exp. Med. 1987, 165, 500–514. [Google Scholar] [CrossRef]
  16. Coyle, J.T. Puttfarcken Poxidative stress, glutamate, and neurodegenerative disorders. Science 1993, 262, 689–695. [Google Scholar]
  17. Murphy, T.H.; Miyamoto, M.; Sastre, A.; Schnaar, R.L.; Coyle, J.T. Glutamate toxicity in a neuronal cell line involves inhibition of cystine transport leading to oxidative stress. Neuron 1989, 2, 1547–1558. [Google Scholar] [CrossRef]
  18. Yoon, S.W.; Kang, S.; Ryu, S.E.; Poo, H. Identification of tyrosine-nitrated proteins in HT22 hippocampal cells during glutamate-induced oxidative stress. Cell Prolif. 2010, 43, 584–593. [Google Scholar] [CrossRef]
  19. Jacewicz, D.; Szkatuła, M.; Chylewska, A.; Dąbrowska, A.; Chmurzyński, L. Coordinate cis-[Cr(C2O4)(pm)(OH2)2]+ cation as molecular biosensor of pyruvates protective activity against hydrogen peroxide mediated cytotoxity. Sensors 2008, 8, 4487–4504. [Google Scholar] [CrossRef]
  20. Kita, E. Model quasi-enzyme compounds of chromium(III) with vitamins B6 and histamine. Pol. J. Chem. 1991, 65, 1933–1940. [Google Scholar]
  21. Jacewicz, D.; Dąbrowska, A.; Banecki, B.; Kolisz, I.; Woźniak, M.; Chmurzyński, L. A stopped-flow study on the kinetics and mechanism of CO2 uptake by chromium(III) complexes with histamine and pyridoxamine. Trans. Met. Chem. 2005, 30, 209–216. [Google Scholar] [CrossRef]
  22. Lay, P.A.; Levina, A. Kinetics and mechanism of chromium(VI) reduction to chromium(III) by l-cysteine in neutral aqueous solutions. Inorg. Chem. 1996, 35, 7709–7717. [Google Scholar] [CrossRef]
  23. Kissner, R.; Nauser, T.; Bugnon, P.; Lye, P.G.; Koppenol, W.H. Formation and Properties of Peroxynitrite as Studied by Laser Flash Photolysis, High-Pressure Stopped-Flow Technique, and Pulse Radiolysis. Chem. Res. Toxicol. 1997, 10, 1285–1292. [Google Scholar] [CrossRef]
  24. Kuban-Jankowska, A.; Knap, N.; Gorska, M.; Popowska, U.; Woźniak, M. Protein tyrosine phosphatase CD45 as a molecular biosensor of hydrogen peroxide generation in cell culture media. Biochem. Biophys. Res. Commun. 2011, 415, 270–273. [Google Scholar] [CrossRef]
  25. Szabelski, M.; Guzow, K.; Rzeska, A.; Malicka, J.; Przyborowska, M.; Wiczk, W. Acidity of carboxyl group of tyrosine and its analogues and derivatives studiem by steady state fluorescencje spectroscopy. J. Photochem. Photobiol. A Chem. 2002, 152, 73–77. [Google Scholar] [CrossRef]
  26. Johanson, M.L.; Correira, J.J.; Yphantis, D.A.; Halvorson, H.R. Analysis of data from the analytical ultracentrifuge by nonlinear least- squares techniques. Biophys. J. 1981, 36, 575–588. [Google Scholar] [CrossRef]
  27. Nagel, J.F.; Parodi, L.A.; Lozier, R.H. Procedure for testing kinetic models of the photocycle of Bacteriorhodopsin. Biophys. J. 1982, 38, 161–174. [Google Scholar] [CrossRef]
  28. Knutson, J.R.; Beechem, J.M.; Brand, L. Simultaneous analysis of multiple fluorescence decay curves: A global approach. Chem. Phys. Lett. 1983, 102, 501–507. [Google Scholar] [CrossRef]
  29. Maeder, M.; Zuberbuchler, A. Nonlinear Least-Squares Fitting of Multivariate Data. Anal. Chem. 1990, 64, 2220–2224. [Google Scholar] [CrossRef]
  • Sample Availability: Samples of the compounds are available from the authors.

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Jacewicz, D.; Siedlecka-Kroplewska, K.; Pranczk, J.; Wyrzykowski, D.; Woźniak, M.; Chmurzyński, L. Cis-[Cr(C2O4)(pm)(OH2)2]+ Coordination Ion as a Specific Sensing Ion for H2O2 Detection in HT22 Cells. Molecules 2014, 19, 8533-8543. https://doi.org/10.3390/molecules19068533

AMA Style

Jacewicz D, Siedlecka-Kroplewska K, Pranczk J, Wyrzykowski D, Woźniak M, Chmurzyński L. Cis-[Cr(C2O4)(pm)(OH2)2]+ Coordination Ion as a Specific Sensing Ion for H2O2 Detection in HT22 Cells. Molecules. 2014; 19(6):8533-8543. https://doi.org/10.3390/molecules19068533

Chicago/Turabian Style

Jacewicz, Dagmara, Kamila Siedlecka-Kroplewska, Joanna Pranczk, Dariusz Wyrzykowski, Michał Woźniak, and Lech Chmurzyński. 2014. "Cis-[Cr(C2O4)(pm)(OH2)2]+ Coordination Ion as a Specific Sensing Ion for H2O2 Detection in HT22 Cells" Molecules 19, no. 6: 8533-8543. https://doi.org/10.3390/molecules19068533

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