The experimental study of the pathogenic role of NADPH-oxidase in the thyroid gland tissues

The objective of the study was to compare the NADPH/ H2O2 system and the expression of enzymes in the culture of thyroid gland cells, depending on the sex. Material and methods. The thyroid gland was collected from adult female donors (n=20) and men (n=15). The cells were washed twice with HBSS and centrifuged at 1000 × g for 5 minutes, and then cultured to 106 cells in 0.5 ml culture medium per well in 24-well plates (Thermo Fisher Scientific, USA). H2O2 levels in the cells were determined quantitatively using a hydrogen peroxide sensor Premo™ Orp1-roGFP (Life Technologies, USA) according to manufacturer’s instructions. Statistical analysis of the results was carried out using Statistica 6.1 (StatSoft, Inc., USA). Results. We found that the expression of DUOX1 and NOX4 in the thyroid gland was significantly higher in women than in men (1.5 times, p<0.05). Expression of DUOX2 in female cells was 1.5 times higher than in men, but statistically significant (p=0.06). We have found that the thyroid gland of women produces a significantly higher H2O2 content than the male gland cells (p<0.05). The expression of DUOX1 and NOX4 is significantly higher in cells derived from the female thyroid gland than in cells derived from males. H2O2 levels were also higher in cells derived from female thyroid gland than in male cells. Conclusion. As a result of our experiment, the expression of DUOX1 and NOX4 is significantly higher in cells derived from the female thyroid gland than in cells derived from males. We have shown during the experiment that NOX4 is regulated both at the transcription level and at the level after transcription, and the level of sustained-release NOX4 mRNA does not accurately reflect the content and function of the NOX4 protein. Chronic effects on the female thyroid gland at higher concentrations of H2O2 can potentially cause many adverse effects

Investigate the Possibility of Using Phosphorescence in Clinical Oncology as an Early Prognostic Test in Detecting Brain Carcinogenesis.

Phosphorescence is considered one of the non-invasive glioblastoma testing methods based on studying molecular energy and the metabolism of L-tryptophan (Trp) through KP, which provides essential information on regulating immunity and neuronal function. This study aimed to conduct a feasibility study using phosphorescence in clinical oncology as an early prognostic test in detecting Glioblastoma. This study was conducted on 1039 patients who were operated on with follow-up between January 1, 2014, and December 1, 2022, and retrospectively evaluated in participating institutions in Ukraine (the Department of Oncology, Radiation Therapy, Oncosurgery, and Palliative Care at the Kharkiv National Medical University). Method of protein phosphorescence detection included two steps. During the first step, of luminol-dependent phosphorescence intensity in serum was carried out after its activation by the light source, according to the spectrofluorimeter method, as follows. At a temperature of 30 °C, serum drops were dried for 20 min to form a solid film. After that, we put the quartz plate with dried serum in a phosphoroscope of luminescent complex and measured the intensity. With the help of Max-Flux Diffraction Optic Parallel Beam Graded Multilayer Monochromator (Rigaku Americas Corporation) following spectral lines as 297, 313, 334, 365, 404, and 434 nm were distinguished and absorbed by serum film in the form of light quantum. The monochromator exit split width was 0.5 mm. Considering the limitations of each of the non-invasive tools currently available, phosphorescence-based diagnostic methods are ideally integrated into the NIGT platform: a non-invasive approach for visualizing a tumor and its main tumor characteristics in the spatial and temporal order. Because trp is present in virtually every cell in the body, these fluorescent and phosphorescent fingerprints can be used to detect cancer in many different organs. Using phosphorescence, it is possible to create predictive models for GBM in both primary and secondary diagnostics. This will assist clinicians in selecting the appropriate treatment option, monitoring treatment, and adapting to the era of patient-centered precision medicine

Transcriptional Dynamics of DNA Damage Responsive Genes in Circulating Leukocytes during Radiotherapy

External beam radiation therapy leads to cellular activation of the DNA damage response (DDR). DNA double-strand breaks (DSBs) activate the ATM/CHEK2/p53 pathway, inducing the transcription of stress genes. The dynamic nature of this transcriptional response has not been directly observed in vivo in humans. In this study we monitored the messenger RNA transcript abundances of nine DNA damage-responsive genes (CDKN1A, GADD45, CCNG1, FDXR, DDB2, MDM2, PHPT1, SESN1, and PUMA), eight of them regulated by p53 in circulating blood leukocytes at different time points (2, 6&ndash;8, 16&ndash;18, and 24 h) in cancer patients (lung, neck, brain, and pelvis) undergoing radiotherapy. We discovered that, although the calculated mean physical dose to the blood was very low (0.038&ndash;0.169 Gy), an upregulation of Ferredoxin reductase (FDXR) gene transcription was detectable 2 h after exposure and was dose dependent from the lowest irradiated percentage of the body (3.5% whole brain) to the highest, (up to 19.4%, pelvic zone) reaching a peak at 6&ndash;8 h. The radiation response of the other genes was not strong enough after such low doses to provide meaningful information. Following multiple fractions, the expression level increased further and was still significantly up-regulated by the end of the treatment. Moreover, we compared FDXR transcriptional responses to ionizing radiation (IR) in vivo with healthy donors&rsquo; blood cells exposed ex vivo and found a good correlation in the kinetics of expression from the 8-hours time-point onward, suggesting that a molecular transcriptional regulation mechanism yet to be identified is involved. To conclude, we provided the first in vivo human report of IR-induced gene transcription temporal response of a panel of p53-dependant genes. FDXR was demonstrated to be the most responsive gene, able to reliably inform on the low doses following partial body irradiation of the patients, and providing an expression pattern corresponding to the % of body exposed. An extended study would provide individual biological dosimetry information and may reveal inter-individual variability to predict radiotherapy-associated adverse health outcomes