Molecular insights into the effect of ozone on human hemoglobin in autohemotherapy: Highlighting the importance of the presence of blood antioxidants during ozonation

Mehraban, Fouad; Seyedarabi, Arefeh; Seraj, Zahra; Ahmadian, Shahin; Poursasan, Najmeh; Rayati, Saeed; Moosavi-Movahedi, Ali Akbar

doi:10.1016/j.ijbiomac.2018.08.028

Cited by 13 publications

(40 citation statements)

References 36 publications

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“…Ozone was generated by passing pure oxygen gas using an electrical corona arc discharge and by a generator made by Gardina Corporation (Iran). The flow and concentration of ozone gas was controlled by photometry (Mehraban et al, 2018). The use of filtered air due to the presence of 78% nitrogen results in the formation of nitrogen oxides which is not desirable in ozone therapy.…”

Section: Ozone Generationmentioning

confidence: 99%

“…Blood was first taken from a healthy person and then Hb purified from it similar to our previous study (Mehraban et al, 2018). In summary, after centrifugation and removal of plasma, the red blood cells were washed with saline and lysed with cold water, followed by addition of ammonium sulfate to precipitate and separate the redundant protein before Hb was dialyzed in phosphate buffer.…”

Section: Collection and Preparation Of Hbmentioning

confidence: 99%

“…The fluorescence spectra were measured, as in our previous study (Mehraban et al, 2018), using excitation at a wavelength of 280 nm and emission in the range of 300 to 400 nm. Protein concentration was determined from standard absorbance measurements using a UV spectrophotometer at 280 nm.…”

Section: Fluorescence Spectroscopymentioning

confidence: 99%

“…Ozone dissolves rapidly in the aquatic environment of the blood and gives its energy to the most hematic portion of the blood, hemoglobin (Hb) (Bocci et al, 2011b). Recently, the effects of ozone on Hb have been extensively studied (Karnaukhova et al, 2014;Mehraban et al, 2018Mehraban et al, , 2019.…”

Section: Introductionmentioning

confidence: 99%

“…Structural changes in protein in the presence of multiple compounds can be explained by their respective interactions (Kamshad et al, 2019;Mokaberi et al, 2019Mokaberi et al, , 2020. In Hb, structural changes have been reported upon ozonation in different environments and under different conditions using multiple spectroscopic techniques (Mehraban et al, 2018(Mehraban et al, , 2019.…”

Section: Introductionmentioning

confidence: 99%

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Highlighting the Importance of Water Alkalinity Using Phosphate Buffer Diluted With Deionized, Double Distilled and Tap Water, in Lowering Oxidation Effects on Human Hemoglobin Ozonated at High Ozone Concentrations in vitro

Mehraban

Rayati

Mirzaaghaei³

et al. 2020

Front. Mol. Biosci.

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In autohemotherapy, it is important to find a way to lower the effects of oxidation, especially at high concentrations of ozone. One of the parameters, other than ozone concentration, which can have a significant effect on the stability and rate of decomposition of ozone at high concentrations, is the presence of ions in water. A number of spectroscopic techniques including intrinsic fluorescence, circular dichroism and UV-VIS were used as well as SDS-PAGE, Native-PAGE dynamic light scattering and water ion analysis, in order to investigate the effects of two relatively high concentrations of ozone on purified human hemoglobin (Hb) in phosphate buffer and diluted versions with deionized, double distilled and tap water in vitro. Purified human Hb and not whole blood human Hb was used in this study, since the addition of water to the whole blood would have caused the RBCs to lyse, affecting the purification of Hb for further analysis. Therefore, using purified Hb, it was possible to investigate the effects of dilution of 50 mM phosphate buffer to 10 mM phosphate buffer with different water types including non-ion containing deionized and double distilled water as well as ion-containing tap water, when ozonated at 55 and 80 µg/ml ozone. The fundamental changes in the secondary and tertiary structures of Hb were seen to be related to non-ozonated Hb samples diluted with deionized and double distilled waters, respectively. Generally, Hb oligomerization was more likely to occur at the higher concentration of ozone (80 µg/ml) and in samples where phosphate buffer was diluted with non-ion containing deionized and double distilled waters and not the ion-containing tap water. This could be explained by the presence of water alkalinity or bicarbonate ions in tap water, which can scavenge free radicals and reduce Hb oxidation/oligomerization. Therefore, it was concluded that

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Section: Ozone Generationmentioning

confidence: 99%

Section: Collection and Preparation Of Hbmentioning

confidence: 99%

Section: Fluorescence Spectroscopymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Highlighting the Importance of Water Alkalinity Using Phosphate Buffer Diluted With Deionized, Double Distilled and Tap Water, in Lowering Oxidation Effects on Human Hemoglobin Ozonated at High Ozone Concentrations in vitro

Mehraban

Rayati

Mirzaaghaei³

et al. 2020

Front. Mol. Biosci.

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The molecular effects of ozone on human hemoglobin oligomerisation pre- and post-COVID-19 infection accompanied by favoured antioxidant roles of cinnamaldehyde and phenyl ethyl alcohol

Mahlooji,

Naderi Beni,

Mehraban

et al. 2025

Journal of Molecular Structure

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A Novel Molecular Approach for Enhancing the Safety of Ozone in Autohemotherapy and Insights into Heme Pocket Autoxidation of Hemoglobin

Naderi Beni,

Hassani-Nejad Pirkouhi,

Mehraban

et al. 2023

ACS Omega

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Major ozone autohemotherapy (MAH) is a popular clinical practice for treating a variety of pathological conditions due to the mild and controlled oxidative stress produced by the reaction of ozone gas with other biological components. Previous studies have shown that blood ozonation leads to structural changes in hemoglobin (Hb); therefore, in the present study, the molecular effects of ozonation on Hb of a healthy individual were assessed by ozonating whole blood samples with single doses of ozone at 40, 60, and 80 μg/mL or double doses of ozone at 20 + 20, 30 + 30, and 40 + 40 μg/mL ozone to investigate whether ozonating once versus twice (but with the same final ozone concentration) would have varying effects on Hb. Additionally, our study aimed to verify whether using a very high ozone concentration (80 + 80 μg/mL), despite mixing it with blood in two steps, would result in Hb autoxidation. The pH, oxygen partial pressure, and saturation percentage of the whole blood samples were measured through a venous blood gas test, and the purified Hb samples were analyzed using several techniques including intrinsic fluorescence, circular dichroism and UV–vis absorption spectroscopies, SDS-polyacrylamide gel electrophoresis, dynamic light scattering, and a zeta potential analyzer. Structural and sequence analyses were also used to study the Hb heme pocket autoxidation sites and the residues involved. The results showed that the oligomerization and instability of Hb can be reduced if the ozone concentration to be used in MAH is divided into two doses. Indeed, our study demonstrated that two-step ozonation with 20, 30, and 40 μg/mL of ozone instead of single-dose ozonation with 40, 60, and 80 μg/mL of ozone reduced the potential adverse effects of ozone on Hb including protein instability and oligomerization. Moreover, it was found that for certain residues, their orientation or displacement leads to the entry of excess water molecules into the heme moiety, which can contribute to Hb autoxidation. Additionally, the autoxidation rate was found to be higher in alpha globins compared to beta globins.

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Molecular insights into the effect of ozone on human hemoglobin in autohemotherapy: Highlighting the importance of the presence of blood antioxidants during ozonation

Cited by 13 publications

References 36 publications

Highlighting the Importance of Water Alkalinity Using Phosphate Buffer Diluted With Deionized, Double Distilled and Tap Water, in Lowering Oxidation Effects on Human Hemoglobin Ozonated at High Ozone Concentrations in vitro

Highlighting the Importance of Water Alkalinity Using Phosphate Buffer Diluted With Deionized, Double Distilled and Tap Water, in Lowering Oxidation Effects on Human Hemoglobin Ozonated at High Ozone Concentrations in vitro

The molecular effects of ozone on human hemoglobin oligomerisation pre- and post-COVID-19 infection accompanied by favoured antioxidant roles of cinnamaldehyde and phenyl ethyl alcohol

A Novel Molecular Approach for Enhancing the Safety of Ozone in Autohemotherapy and Insights into Heme Pocket Autoxidation of Hemoglobin

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