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The increase in the integral intensity of the EPR signal from Fe³⁺ ions in transferrin after the delivery of the radiation dose. EPR signal in a patient before (solid line) and after (dashed line) radiation exposure
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Ceruloplasmin and transferrin are proteins which play a potential role in the process of breast cancer development. These molecules contain Cu²⁺ (ceruloplasmin) or Fe³⁺ ions (transferrin) and thus constitute paramagnetic centers, which can be studied using electron paramagnetic resonance method. The aim of the study was to determine how paramagneti...
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Purpose: to reveal protective effects of intravenous low-level laser blood irradiation (ILBI) during surgical interventions at the gallbladder as well as to study the state of antioxidant activity, lipid peroxidation and endogenous intoxication.
Material and methods. In the perioperative period, 40 patients from the main group (I) had traditional t...
Citations
... The only known direct method that enables the measurement of free radicals is electron paramagnetic resonance (EPR) [20]. This method has been successfully applied in previous studies on free radicals in biological materials, such as blood samples [21][22][23], and in the studies of radical processes occurring in both healthy and cancer cells [24][25][26]. ...
The effect of nanosecond electromagnetic pulses on human health, and especially on forming free radicals in human cells, is the subject of continuous research and ongoing discussion. This work presents a preliminary study on the effect of a single high-energy electromagnetic pulse on morphology, viability, and free radical generation in human mesenchymal stem cells (hMSC). The cells were exposed to a single electromagnetic pulse with an electric field magnitude of ~1 MV/m and a pulse duration of ~120 ns generated from a 600 kV Marx generator. The cell viability and morphology at 2 h and 24 h after exposure were examined using confocal fluorescent microscopy and scanning electron microscopy (SEM), respectively. The number of free radicals was investigated with electron paramagnetic resonance (EPR). The microscopic observations and EPR measurements showed that the exposure to the high-energy electromagnetic pulse influenced neither the number of free radicals generated nor the morphology of hMSC in vitro compared to control samples.
... It is based on the interaction of unpaired electrons (free radicals) with a magnetic field [10,11]. ESR measurement provides information about the concentration of free radicals (quantitative data) and the type of free radicals (qualitative data) [12][13][14]. Free radical concentration is derived from ESR signal intensity, proportional to double integral. A free radical is determined based on a given radical's hyperfine pattern and g-factor value. ...
Spin probes can be used to monitor biological membranes, including the penetration of different molecules into cells. The aim of the present studies was an investigation of the endocytosis process of two spin labels—2,2,6,6-Tetramethylpiperidine-1-oxyl (TEMPO) and 4-hydroxy-TEMPO (TEMPOL)—into yeast cells and a leukemia cell line (HL-60, ATCC CCL-240) by Electron Spin Resonance (ESR). The ESR method is helpful for the direct detection of free radicals. The cell incubation and endocytosis of spin probes were carried out at 310 K. In contrast, the ESR measurements of yeast cells and a leukemia cell line with spin probes were at 240 K. Spectral differentiation was observed; hence, the spin probes present in suspension and attached to the cell membrane could be distinguished. The ESR signal changes of spin probes depended on spin probe concentration, cell number, and type of cell (healthy/cancerous). Additionally, the effect of external factors (oxygen and vitamin C) on the ESR signal decay of spin markers in the cell solution was established. The experimental results prove that the spin probes (TEMPO and TEMPOL) could scavenge free radicals inside the cell. At the same time, the mechanism of spin probe interaction in suspension was determined based on the measurements at low temperatures.
... Cp is a member of the multicopper oxidase family of enzymes. Previous reports showed that Cp is not only synthesized in the liver, but also produced by cancer cells [54]. Tumor cells can capture non-Cp copper from plasma to form pathological angiogenesis, which need a relatively large amount of copper; therefore, the expression level of Cp might represent a supportive biomarker for bile duct cancer and breast cancer [54,55]. ...
... Previous reports showed that Cp is not only synthesized in the liver, but also produced by cancer cells [54]. Tumor cells can capture non-Cp copper from plasma to form pathological angiogenesis, which need a relatively large amount of copper; therefore, the expression level of Cp might represent a supportive biomarker for bile duct cancer and breast cancer [54,55]. In our study, we evaluated the diagnostic value of PHA-E-positive Cp for PC by drawing ROC curves. ...
Pancreatic cancer (PC) remains one of the top 10 causes of cancer-related death in recent years. Approximately 80% of PC patients are diagnosed at the middle or advanced stage and miss the opportunity for surgery. The demand for early diagnostic methods and reliable biomarkers is increasing, although a number of tumor markers such as CA19-9 and CEA have already been utilized in clinics. In this study, we analyzed the alteration of N-glycan of serum glycoproteins by mass spectrometry and lectin blotting. The results showed that bisecting GlcNAc structures of glycoproteins are significantly increased in PC patients’ sera. With Phaseolus vulgaris Erythroagglutinin (PHA-E) lectin that specifically recognizes bisecting GlcNAc N-glycans, the serum glycoproteins bearing bisecting GlcNAc in PC patients’ sera were pulled down and identified by nano-LC-MS/MS. Among them, ceruloplasmin (Cp) was screened out with a satisfied sensitivity and specificity in identifying PC from acute pancreatitis patients (AUC: 0.757) and normal healthy persons (AUC: 0.972), suggesting a close association between Cp and PC development and diagnosis. To prove that, the Cp expression in tumor tissues of PC patients was examined. The results showed that Cp was significantly upregulated in PC tissues compared to that in adjacent normal tissues. All these results suggested that PHA-E-positive Cp could be a potential PC-specific glycoprotein marker to distinguish PC patients from acute pancreatitis patients and normal persons.
... Based on our findings, we hypothesize that CP downregulation after irradiation either i) reflects cellular stress or, alternatively, ii) enables GB cells to better resist radiation. Interestingly, a recent study showed reduced CP in whole blood of patients with breast cancer after delivery of radiation [46]. Since radiotherapy aims to eradicate tumor by inducing cancer cell death, its potential role could be assigned to different functions such as: i) prevention of damage by deregulating the cycle of iron and therefore its cellular availability and by extension its contribution in the Fenton reaction and ROS production, or ii) a direct link with DNA damage or increase the capacity of repair machinery. ...
Background
Glioblastoma (GB) is the most common and most aggressive malignant brain tumor. In understanding its resistance to conventional treatments, iron metabolism and related pathways may represent a novel avenue. As for many cancer cells, GB cell growth is dependent on iron, which is tightly involved in red-ox reactions related to radiotherapy effectiveness. From new observations indicating an impact of RX radiations on the expression of ceruloplasmin (CP), an important regulator of iron metabolism, the aim of the present work was to study the functional effects of constitutive expression of CP within GB lines in response to beam radiation depending on the oxygen status (21% O2 versus 3% O2).
Methods and results
After analysis of radiation responses (Hoechst staining, LDH release, Caspase 3 activation) in U251-MG and U87-MG human GB cell lines, described as radiosensitive and radioresistant respectively, the expression of 9 iron partners (TFR1, DMT1, FTH1, FTL, MFRN1, MFRN2, FXN, FPN1, CP) were tested by RTqPCR and western blots at 3 and 8 days following 4 Gy irradiation. Among those, only CP was significantly downregulated, both at transcript and protein levels in the two lines, with however, a weaker effect in the U87-MG, observable at 3% O2. To investigate specific role of CP in GB radioresistance, U251-MG and U87-MG cells were modified genetically to obtain CP depleted and overexpressing cells, respectively. Manipulation of CP expression in GB lines demonstrated impact both on cell survival and on activation of DNA repair/damage machinery (γH2AX); specifically high levels of CP led to increased production of reactive oxygen species, as shown by elevated levels of superoxide anion, SOD1 synthesis and cellular Fe2 + .
Conclusions
Taken together, these in vitro results indicate for the first time that CP plays a positive role in the efficiency of radiotherapy on GB cells.
... However, access to high-frequency EPR facilities is only rarely available and this is an experimentally challenging methodology. Being so, and despite its limitations, by allowing a more routine use X-band EPR has proven a valuable tool in iron biology studies [118][119][120][121]. ...
Transferrin (Tf) is an essential protein, probably ubiquitous to all metazoans. The main function of this family of proteins is to bind and transport ferric ions, increasing Fe(III) solubility under physiological conditions and preventing its deleterious pro-oxidant role in aerobic environments. In humans, Tf is responsible for the safe transport of this essential micronutrient through circulation and its delivery to requiring cells. Cellular uptake occurs through endocytosis mediated by the transferrin receptor (TfR). Although there is a good understanding of the general molecular mechanisms governing Tf iron binding and cellular iron delivery, several aspects of Tf biochemistry remain unclear.
In this review, we provide an overview of the inorganic biochemistry of human Tf (hTf), exploring the available structural information on hTf and the hTf/TfR complex to discuss physiologically relevant aspects, such as iron binding site distribution, mechanism of iron loading and TfR priming of iron release in vivo. In addition, we consider the role of hTf microheterogeneity on its function. Post-translational modifications such as phosphorylation, glycation and oxidation may occur at relevant hTf sites, which will compromise iron binding, inter lobe cooperativity or interaction with the TfR. Furthermore, these modifications may contribute to the deregulation of systemic iron transport and distribution on a disease specific manner. Finally, a brief review of the role of hTf as one of the main blood serum ligands for toxic and therapeutic metal ions is presented.
hTf is a long known and studied protein, but it is essential to understand the factors governing its expression and catabolism and how specific modifications modulate its function, in order to fully comprehend its role in human pathology or to explore its potential as a therapeutic agent.
... For more information about the Conference please visit the websites: http://ihicps.com/ C h e m i s t r y | 69 cancer has two main types: non-invasive breast cancer and invasive breast cancer [5] .Ceruloplasmin (Cp) is an enzyme that may play a significant role in the progression of breast cancer [6]. The Cp is an acute-phase protein that is typically produced by hepatic cells [7] and is primarily secreted in the plasma [8], whereas Cp mRNA was found in cell lines of breast cancer patients [9]. ...
Ceruloplasmin is considered the main copper transport protein which is proposed to have a role in cancer. Ceruloplasmin is an acute phase reactant and antioxidant enzyme, has been found to be increased in sera of patients with several types of cancers including breast cancer.
The aim of present study was to determine of ceruloplasmin oxidase activity, specific activity, iron concentration in sera of patients with breast cancer and comparing with healthy group, and the ability of using enzyme as a tumor marker for breast cancer.
This study was performed from November 2018 to January 2019, blood samples were collected from breast cancer patients in Nanakeli Hospital in Erbil city. Study was included (65) female patients with breast cancer and (20) healthy donors as control group. Ceruloplasmin activity, specific activity, serum total protein concentration, serum total iron concentration levels were estimated for all samples by colorimetric methods.
The results shown that there was a significant increase in ceruloplasmin activity level for patients with breast cancer ( 227.8 U/L) compared with control group (101.1 U/L) while total protein level for breast cancer patients (6.592 g/dl) showed no significant change compared to control group(7.127 g/dL) likewise the result shown that there was a significant increase in specific activity of ceruloplasmin and total iron concentration level for patients with breast cancer (34.74 U/mg) (334.2 mg/dl) compared to control group (14.14 U/mg) (128.6 mg/dl) The conclusion of this study was the concentration of ceruloplasmin activity, serum total iron concentration and specific activity of ceruloplasmin significantly increase in breast cancer patients while total protein level revealed no significant change.
Serum ceruloplasmin oxidase activity and specific activity were highly significant elevated in patient group when compared with control group. Also the total iron concentration results showed significant increasing (P˂0.0001) between of patient with breast cancer and control group.
Key words: Ceruloplasmin Oxidase Activity, Breast Tumor, Iron.
... Intracellular iron export is mediated by ferroportin (FPN) [65]. FPN exports Fe 2+ to the extracellular space where it is oxidized by ceruloplasmin (CP) [66], hephaestin (HEPH) [67], and zyklopen (HEPHL1) [66]. FPN is expressed in numerous cell types, in particular, in those involved in the regulation of plasma iron levels, such as enterocytes, macrophages, and hepatocytes [68]. ...
... Intracellular iron export is mediated by ferroportin (FPN) [65]. FPN exports Fe 2+ to the extracellular space where it is oxidized by ceruloplasmin (CP) [66], hephaestin (HEPH) [67], and zyklopen (HEPHL1) [66]. FPN is expressed in numerous cell types, in particular, in those involved in the regulation of plasma iron levels, such as enterocytes, macrophages, and hepatocytes [68]. ...
New insights into the field of iron metabolism within the tumor microenvironment have been uncovered in recent years. Iron promotes the production of reactive oxygen species, which may either trigger ferroptosis cell death or contribute to malignant transformation. Once transformed, cancer cells divert tumor-infiltrating immune cells to satisfy their iron demand, thus affecting the tumor immunosurveillance. In this review, we highlight how the bioavailability of this metal shapes complex metabolic pathways within the tumor microenvironment and how this affects both tumor-associated macrophages and tumor-infiltrating lymphocytes functions. Furthermore, we discuss the potentials as well as the current clinical controversies surrounding the use of iron metabolism as a target for new anticancer treatments in two opposed conditions: i) the "hot" tumors, which are usually enriched in immune cells infiltration and are extremely rich in iron availability within the microenvironment, and ii) the "cold" tumors, which are often very poor in immune cells, mainly due to immune exclusion.
... Ceruloplasmin loss-of-function is involved in neurodegenerative syndrome, whereas high ceruloplasmin level is associated with copper toxicity, Alzheimer's disease and cardiovascular diseases 43,44 . As ceruloplasmin levels were significantly high in cancer, ceruloplasmin has been suggested to be potential biomarkers in numerous solid tumors [45][46][47][48][49][50] . The relationship between ceruloplasmin level and bladder cancer has not been documented previously. ...
The identification of clinically-relevant early diagnostic and prognostic protein biomarkers is essential to maximize therapeutic efficacy and prevent cancer progression. The aim of the current study is to determine whether aberrant plasma protein profile can be applied as a surrogate tool for early diagnosis of bladder carcinoma. Plasma samples from patients with low grade non-muscle invasive bladder cancer and healthy controls were analyzed using combined 2D-DIGE and mass-spectrometry to identify differentially expressed proteins. Validation was performed using western blotting analysis in an independent cohort of cancer patients and controls. Fifteen differentially-expressed proteins were identified of which 12 were significantly up-regulated and three were significantly down-regulated in tumors compared to controls. The Ingenuity Pathways Analysis revealed functional connection between the differentially-expressed proteins and immunological disease, inflammatory disease and cancer mediated through chemokine and cytokine signaling pathway and NF-kB transcription factor. Among the three validated proteins, haptoglobin was able to distinguish between patients with low grade bladder cancer and the controls with high sensitivity and specificity (AUC > 0.87). In conclusion, several biomarker proteins were identified in bladder cancer. Haptoglobin is a potential candidate that merit further investigation to validate its usefulness and functional significance as potential biomarkers for early detection of bladder cancer.
... Electron spin resonance (ESR) is commonly used for characterization of physical properties of various nanomaterials [22][23][24][25][26][27][28] , including functionalized magnetic nanoparticles 29,30 . The magnetic nanoparticles with attached spin label (free radical) are characterized by two areas that can be described by ESR measurements -magnetic core and free radical. ...
Potential application of magnetic nanoparticles as drug carriers in medical treatment requires prior determination of their effects on cells. In this work different spin labels and magnetic nanoparticles functionalized with spin labels as well as their interaction with yeast cells were investigated using electron spin resonance (ESR) method. ESR was demonstrated to be a suitable method for monitoring of magnetic core and attached spin labels. Particular emphasis was placed on characterization of endocytosis and redox processes running inside the cell, resulting in recombination of spin labels. Such data could only be obtained at reduced temperature of ESR measurements.
... Thus, it has been shown that an increase in Cu/Zn ratio in TT and serum of patients with hormone-dependent tumors evidenced on activation of matrix metalloproteinases that are involved in the processes of invasion and metastasis [18,21]. Along with this, it was found that the change in Cu/Zn ratio may be due to the disruption of the activity of some enzymes of the AOS [21,37,38]. It is also noteworthy that the formation of an invasive phenotype of tumor cells in breast cancer is accompanied by an increase of Cu/Mg ratio in the serum of these patients. ...
Aim:
To investigate the content of essential elements (EE): copper, zinc, magnesium, iron and calcium and the evaluation of the activity of metal-containing enzymes - ceruloplasmin (CP), myeloperoxidase (MPO) and the content of transferrin (TF) in blood plasma (BP) and tumor tissue (TT) of animals with Walker-256 carcinosarcoma treated with lactoferrin (LF).
Materials and methods:
The study of the EE content and the activity of the abovementioned enzymes was carried out on rats with Walker-256 carcinosarcoma treated with LF at the doses of 1 and 10 mg/kg of body weight. The quantitative content of EE in BP and TT of animals was determined using the inductively coupled plasma atomic emission spectroscopy (ICP-AES). Determination of CP activity, content of TF and hemochromes was performed using the method of electron paramagnetic resonance (EPR), and MPO - by unified biochemical method.
Results:
The introduction of LF at the doses of 1 and 10 mg/kg resulted in a decrease in the ratio of Cu/Zn in BP and even more expressed decrease of Ca/Mg ratio in TT. Administration of LF, especially at a dose of 10 mg/kg, affected the increase in CP and MPO activity in BP. It has been shown that administration of LF at a dose of 10 mg/kg led to an increase in oxidative products of destruction of the hemoglobin-hemochrom system in the TT, against the background of lowering the TF content.
Conclusions:
The administration of LF, especially at a dose of 10 mg/kg, led to metabolic alterations associated with inhibition of the tumor process. The detected modulating effect of LF on the content of the EE and the activity of the CP and MPO may be a basis for correction of the elemental balance in carcinogenesis.
