Investigate the possibility of using phosphorescence in clinical oncology as an early prognostic test in detecting brain carcinogenesis

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Vinnyk, Igor A. Kryvoruchko, Valeriy V. Boyko, Yulia V. Ivanova, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-2665331/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 27 Apr, 2023 Read the published version in Journal of Fluorescence → Version 1 posted 7 You are reading this latest preprint version Abstract 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. Materials and Methods. 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 o C, serum drops were dried for 20 minutes 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. Results and conclusion. 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. phosphorescence non-invasive glioblastoma testing L-tryptophan cancerous brain tissues. Figures Figure 1 Figure 2 1. Introduction Glioblastoma (GBM) is the most aggressive primary brain tumor, with an incidence of 2–3 cases per 100,000 people [1]. The average survival in Ukraine of about 12-14 months is achieved with intensive multimodal treatment, but the number of such patients is minimal. Despite such intensive treatment, there is no cure, and the recurrence of Glioblastoma is inevitable [2]. Approaches to diagnosing GBM are rapidly evolving and are based on the recently revised WHO (2016) criteria for classifying central nervous system tumors [3]. Currently, histopathological examination of a tissue sample for suspected GBM is the gold standard for diagnosis. Now complemented by molecular diagnostics, the identification of O6-methylguanine-DNA methyltransferase (MGMT) methylation, isocitrate dehydrogenase (IDH) mutation, and 1p19q coding is currently the most valuable in daily clinical practice [4]. Several studies have proven that cellular metabolism is the main regulator of tumor behavior [5]. To maintain a high proliferation rate, cancer cells undergo metabolic reprogramming, and tumors use amino acids to meet energy needs and biosynthesis in cells [6]. Tryptophan plays an essential role in malignant conversion and tumor progression. Trp degradation occurs via the kynurenine (Kyn) pathway, followed by the formation of metabolites [7]. Non-invasive glioblastoma testing (NIGT) combines non-invasive (non-surgical) techniques to represent the tumor and provides information about tumor drivers and the microenvironment, all factors that can/should be included in a treatment regimen. Among the large number of NIGTs using spectroscopic methods available for testing cell populations and the functional state of biological fluids, the phosphorescence method occupies a leading position. Phosphorescence spectroscopy of amino acids such as Tryptophan is a fast, easy-to-use, highly sensitive, and highly selective method that allows you to gain in-depth knowledge of the structural, functional, and metabolic changes in cells, organs, and systems that occur at the molecular level [8, 9, 10]. Phosphorescence is considered one of the NIGT 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 [11, 12–16]. This study aimed to conduct a feasibility study using phosphorescence in clinical oncology as an early prognostic test in detecting Glioblastoma. 2. Materials And Methods 2.1. Patient selection and data collection 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). The respective institutional review boards of each participating institution have approved this study. Demographic and clinical data were collected, including age, gender, concurrent diseases, MRI, white blood cell count, and total neutrophil and lymphocyte count. This retrospective cohort study was handled in accordance with the Declaration of Helsinki. This manuscript adheres to the applicable STROBE guideline. The use of registered data follows the General Data Protection Regulation of the European Union. The patient’s written Informed consent was signed for each bio-object from the residual materials. The study and the use of data were consented to by the Ethics Committee of Kharkiv National Medical University, Ukraine (Protocol No. 6, November 11, 2022). Participants must meet one of the following: diagnosed with a single primary cancer that has not yet been treated. There is no evidence or treatment of any cancer for at least five years before enrollment. Key Exclusion Criteria: a medical condition which, in the investigator's opinion, should preclude enrollment in the study. Known to be pregnant. Any therapy for cancer, including surgery, chemotherapy, immunotherapy, and/or radiation therapy, in the five years, preceding enrollment. Participated in or is currently participating in a clinical research study in which an experimental medication has been administered in the last 30 days. Participated in or is currently participating in another clinical study. For the control cohort: Any previous cancer diagnosis in the five years preceding enrollment, recurrence of the same primary cancer within any timeframe, or concurrent diagnosis of multiple primary cancers within any timeframe. The comparative group consisted of 486 conventionally healthy men and women aged from 25 to 65 years. Serum proteins' phosphorescence state was studied and evaluated in 1039 patients with cancer brains (Glioblastoma), aged from 35 to 68 years. Clinical and histomorphological methods confirmed the diagnosis. Among 1039 patients were 764 (406 men, 358 women) patients with Glioblastoma (GB) and 275 (150 men and 125 women) with Multiforme Glioblastoma (MGB). After detecting a brain tumor on CT or MRI, the neurosurgeon obtained tissue from the tumor for biopsy. We used tumor tissue analysis to assign tumor, severity and histological grade. The first stage (I) of the disease was diagnosed in 81 patients (these were cases of cystic brain disease), the second (II) in 163, the third (III) in 564, and the fourth stage (IV) was detected in 231 patients. 2.2 Method of protein phosphorescence detection included two steps. During the first step, we obtain a blood sample (5.0 ml) from the ulnar vein and prepare serum by centrifugation using routine methods in medical practice. A study of luminol-dependent phosphorescence intensity in serum was carried out after its activation by the light source, according to the spectrofluorimeter method, as follows: 50 mcl of luminol-containing serum (10 mcl 3% luminol solution) was placed on a 5 x 45 mm quartz plate and placed in a thermostat. At a temperature of 30 o C, serum drops were dried for 20 minutes 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. The spectral sensitivity of the FEP was located in the ultraviolet and visible ranges of daylight. Coating technology provides results in less than 0.5% d-spacing variation. In the second stage, we examined the tumor tissue after surgery. Fresh surgical specimens of cancerous brain tissues were acquired under Ukraine Association of Biobank approval from a tissue bank within three h of surgery. The standard tissue specimens were from the tumor's margins and were a mix of malignant and normal tissues. Samples were stored at 4°C and not subjected to additional processing before measuring the spectra. Specimens were cut into pieces ranging from ∼0.5 to ∼1 cm 2 and placed in a 1- x 1-cm quartz cuvette before mounting on the translation stage. Using the spectrograph setup, the spectra were acquired in a grid pattern with a 0.5 - to 1-mm spacing to cover most of the specimen. Each spectrum was integrated for 2s. Intensity ratio maps were generated from the individual spectra. All measurement procedures were automated, and the error was not more than 3% in all cases. 2.3 Statistical analysis The summary statistics for continuous variables were calculated separately for each screening group by the number of subjects with the number of recorded observations (N), mean, standard deviation (SD), minimum (Min), first quartile (Q1), median, third quartile (Q3), and maximum (Max). Categorical data were displayed by the screening group using absolute frequencies and percentages. For continuous variables, a between-group comparison will be performed using the Student's t-test for independent samples or the Wilcoxon test, depending on the results of checking the normal distribution of data in groups using the Shapiro-Wilk test. Group comparisons will be made using Pearson's chi-squared test (χ2) for categorical variables. If, in any of the cells of the contingency table, the expected frequencies were less than 5, Fisher's exact test would be applied for comparison. Statistical processing of the obtained data was carried out using the software package Statistica v. 6.0 (Statsoft Inc.). 3. Results The main difference in the levels of serum phosphorescence between patients and conditionally healthy people was observed at the activation of a 404 nm spectral wave. So, the intensity of serum phosphorescence in cancer patients raised 3.6 and 3.8 times, respectively, in men and women. This wave of activation was appropriate for hemoglobin. It can be indicated the loss of compact structure and functional activity by this protein that may result from tissue hypoxia. Research of serum phosphorescence intensity in patients with brain cancer according to the type of tumor process revealed similar dynamics in all studying groups (Table 1). Table 1 The intensity of luminol-dependent serum phosphorescence in patients with brain cancer and conditionally healthy people depends on tumor type The range of activation (nm) Phosphorescence intensity ( І 0 с), М±m c2/P value Patients with Glioblastoma (n=1039) Conditionally healthy people (n=486) MGB (n=275) MG (n=764) 297 5997,8±364,5* 6731,4±356,7* 1.376/0.503 1.376/0.503 313 437,6±65,7* 483,5±76,8* 12.469/0.014 12.469/0.014 334 807,4±53,6* 796,3±68,4* 1.179/0.555 1.179/0.555 365 3014,5±186,3* 2868,7±154,3* 8.752/0.013 8.752/0.013 404 1894,3±145,8* 1812,4±133,6* 0.369/0.832 0.369/0.832 434 1826,4±103,7* 1938,6±145,2* 0.927/0.629 0.927/0.629 The highest level of phosphorescence was observed at activation with a 297 nm spectral wave. In this case, serum phosphorescence raised at 86.8 % and 109.6% in patients with GB and MGB. The intensity of serum phosphorescence at activation with the spectral line of 404 nm was increased by 3.8 and 3.6 times, or by 271.4% and 258.6%, respectively, in patients with GB and MGB compared to a group of conditionally healthy people. The smaller difference in phosphorescence was observed at activation with spectra of 313 nm, 334 nm, and 365 nm. So, at 313 nm, phosphorescence intensity raised at 29.2 %, and 52.1%, at 334 nm – at 26.7 %, and 21.7 %, at 365 nm – at 67.2 %, and 62.9 % accordingly in patients with GB and MGB. The study of serum phosphorescence in GB patients revealed a high informativeness of indexes at activation with a spectral range of 297 nm, 404 nm, and 434 nm. It revealed a direct correlation between the degree of disease severity and the intensity of phosphorescence in GB patients' serum (Table 2). Table 2 The intensity of luminol-dependent serum phosphorescence in patients depending on the stage of GB and MGB The range of activation (nm) Phosphorescence intensity ( І 0 с) , М±m Stage of GB and MGB, n CHP (n=486) P -value Stage - I n= 81 Stage – II n=163 Stage – III n=564 Stage – IV n=231 297 5706,3±127,5* 6157,8±104,5* 6508,3±118,6* 6837,4±162,7* 3210,7±137,5 <0.001 313 394,8±23,6* 415,6±32,7* 488,9±42,7* 520,6±43,8* 317,8±19,6 <0.001 334 762,5±33,8* 794,3±26,5* 825,6±53,2* 863,7±29,4* 653,9±39,6 <0.001 365 2796,8±75,3* 2837,6±83,4* 2925,8±77,4* 3097,6±105,8* 1760,5±59,2 <0.003 404 1754,6±56,3* 1814,3±76,8* 1897,2±83,6* 1972,4±63,5 505,3±39,4 <0.001 434 1710,3±64,5* 1794,8±73,6* 1923,8±105,4* 1986,7±84,3* 593,6±30,9 <0.002 The correlation coefficients in all cases were between 0.87 (r = 0.87) and more. Levels of serum phosphorescence intensity in patients with GB and MGB in activation with 297 nm increased at 77.7 %, 91.7 %, 102.3 %, and 112.3 %, in 313 nm – at 24.2 %, 30.7 %, 53.8 %, and 63.8 %, in 334 nm – at 16.6 %, 21.4 %, 26.2 %, and 32.0 %, in 365 nm – at 58.8 %, 61.2 %, 66.2 %, and 75.9 %, in 404 nm – at 247.2 %, 259.5 %, 275.0 %, and 290.3 %, in 434 nm – at 188.1%, 102, 3%, 224.1 %, and 234.6 %. In all cases, it was noted that phosphorescence intensity rose by more than 200 percent in activation with spectra of 404 nm and 434 nm. And also, a significant increase in phosphorescence was detected at activation with a 297 nm spectral wave. Inactivation of serum with the lower ultraviolet (365 nm) phosphorescence increased by more than 60% compared to a group of conditionally healthy people. In all groups and at different spectral lines, a correlation between phosphorescence intensity and the stage of the tumor process took place. The phosphorescence spectra of D-L-trp powder excited at 282, 300, 380, and 400 nm and acquired with the CD-Scan cancerous brain tissues are shown in Figure 1. Each spectrum shown in the figure is the sum of seven ranges, obtained with delays of 1 to 7 ms in 1-ms steps. The gate width was 1 ms. Thus, each range shown is the integrated intensity from t = 0.5 to 7.5 ms. Although it is well known that the trp excitation maximum is at 282 nm and that trp does not have significant absorption at wavelengths longer than 300 nm, the phosphorescence intensity with 282 nm excitation was weaker than with 400 nm excitation. The phosphorescence was blue-shifted for the longer wavelength excitations (380 and 400 nm) compared to the shorter wavelength excitations (282 and 300 nm). For 380 nm and 400 nm excitation, trp exhibits two phosphorescence peaks at 480 nm and 525 nm. For excitation at 282 nm and 300 nm, only the phosphorescence peak at 525 nm was observed. The mechanism responsible for the shift in phosphorescence and the reason for the more intense at 400 nm excitation is unknown. It may be due to dimers or trimers present in the trp powder. Phosphorescence from ex vivo human brain tissues . Although the phosphorescence intensity in trp powder was greater for excitation at 400 nm than for excitation at 300 nm, tissues contain several fluorophores (NADH and flavins) whose emission wavelengths can overlap with the trp. Therefore, the brain tissue specimens were excited at 300 nm to reduce the contribution from these fluorophores. Figure 2 shows the phosphorescence from ex vivo human normal and malignant brain tissues acquired with the CD-Scan. The plots shown in the figure are each the integration of seven spectra obtained with gate delays of 1 to 7 ms with 1-ms intervals. The normal tissues exhibited phosphorescence emission from 440 to 500 nm. Fluorescence can also be observed at 350 nm — most likely due to a long “tail” on the lamp emission. The normal tissues showed more excellent fluorescence and phosphorescence than the malignant tissues, which had almost no detectable phosphorescence. 4. Discussion Optical biopsy—using native tissue fluorescence—for detecting cancer has been an area of investigation for over two decades. The bases behind the operation of optical biopsy are that the onset of carcinogenesis results in structural changes (thickening of the mucosa layer, increased vascularity) and molecular changes (increased nucleic acids, alterations in protein structure, increased cell metabolism) that modify the spectroscopic properties of tissue and, thereby, create unique optical signatures that can be used to detect malignant and premalignant tissues. In the UV and blue spectral regions, the primary native tissue fluorophores are Tryptophan (trp), collagen, elastin, reduced nicotinamide adenine dinucleotide (NADH), and flavins [ 17 , 18 ]. Optical biopsy has been demonstrated to be an accurate, real-time tool for distinguishing normal tissues from malignant and premalignant tissues. Prior investigations have demonstrated that several ratio-based algorithms can distinguish malignant tissue from normal tissue with high sensitivity and specificity. The wavelength combinations used in those ratios isolated the contributions from different pairs of tissue fluorophores, one of which was trp. The ratio of 340 nm to 440 nm emission (with 300 nm excitation) is useful for identifying malignant tissues from many different organ sites [ 19 ]. According to the findings, the appearance of many molecules in the triplet state in the long-wavelength spectral region of activation may indicate a dissociation of oxidative phosphorylation. The tissue respiration, inhibition of bioenergy processes, which are accompanied by ineffective use of energy and scattering it in the form of heat, and reduced ATP production are symptoms of mitochondrial pathology [ 21 , 22 – 24 ]. The presence of high energy levels in activated electronic states due to the appearance of unpaired electrons in the active molecule indicates a change in the reaction ability and conformational properties of proteins, nucleic acids, and other biologically active molecules (enzymes, hormones). It is known that ultraviolet spectrum photons are absorbed mainly by aromatic amino acids (tyrosine - at 280 nm, Tryptophan - at 220 nm), proteins (at 280–300 nm), nucleic acids, and nucleotides (at 260 nm) which may be present in the serum. An analysis of the results reveals that profound metabolic, structural, and conformational changes of large polymer molecules and their monomeric components occurred in patients with brain cancer. Increased serum phosphorescence intensity in the long-wave region (404–434 nm) may indicate an increase in non-erythrocytic hemoglobin levels and violations in its compact structure, conformational properties, and hemin content (404 nm) as a result. The development of free radical-membrane pathology in patients with brain cancer underpins the formation of hypochromic anemia in carcinogenesis. The study of serum phosphorescence intensity in patients with MGB and MG detected violations of the structural and conformational properties of biologically important macromolecules (proteins, nucleic acids, hemoglobin, and other glycoproteins). The presence of many electron-activated molecules, which can stimulate free radical processes and disconnect oxidative phosphorylation, causes energy deficiency, tissue hypoxia, and membrane molecular pathology, all of which are pathogenic factors in brain carcinogenesis [ 25 , 26 ]. Levels of luminol-dependent serum phosphorescence intensity at activation with 297, 404, and 434 nm can be used to diagnose disease severity and the state of bioenergy processes and plan surgery and pathogenetic therapy. Such spectral monochromatic activation waves as 297, 404, and 434 nm are the most informative. They indicate a disturbance in serum proteins' compact structure and conformational properties and decreased biological activity. High levels of phosphorescence intensity in the first stage of carcinogenesis show changes in the compact design and biological activity of proteins for quite a long period, which precedes the development of the tumor process and is a prognostic factor in the development of oncopathology and its early diagnostic criteria [ 27 – 29 ]. Prolonged activation of oxidative processes leads to changes in protein conformation and biological activity observed in patients with brain cancer. The appearance in the long-wavelength spectral region of activation (404 and 434 nm) and the increased number of molecules in the triplet state may also indicate disconnection of oxidative phosphorylation and tissue respiration, which is always accompanied by heat energy scattering and mitochondrial pathology development, which are pathogenetic factors in carcinogenesis. Increased serum phosphorescence intensity in the ultraviolet region of 297 nm indicates the presence of high levels of triplet-activated states. It shows the change in proteins and nucleic acids' compact structures. The increase in serum phosphorescence in brain cancer patients in the long-wave spectral region of activation (404 nm) reflects an increase in free hemins (a nonprotein component of hemoglobin), confirming the protein's loss of compact structure and biological activity. Informative monochromatic spectral waves of activation in evaluating serum compact structure and biological activity were 297, 404, and 434 nm. Physiological indexes of luminol-dependent serum phosphorescence intensity at activation with 297 nm range from 3000 to 3500 imp/sec. Increasing luminol-dependent serum phosphorescence intensity at activation with 404 nm from 3500 to 5000 and with 434 nm from 400 to 1700 imp/s indicates a conformational change in the structure and biological activity of serum proteins; this can be a significant prognostic indicator in the early diagnosis of the precancerous metabolic state that leads to carcinogenesis and stage-dependent pathological processes. A further rise in luminol-dependent serum phosphorescence intensity from 5000 to 7000 imp/s at 297 nm and from 1700 to 2000 imp/s at 404 and 434 nm indicates an increase in the severity of the disease. This newly developed optical technique for cancer detection, based on phosphorescence and fluorescence spectroscopy, is fast, minimally invasive, and non-destructive. This technique may apply to in vivo or ex vivo tissue analysis. The 345 ∕ 500 ratio, with excitation at 300 nm, provides an excellent fingerprint for cancer detection in ex vivo tissues. A 345 ∕ 500 ratio of 0.5 to 12 corresponds to normal brain tissue, and a ratio higher than 12 indicates cancerous brain tissue. Using ratios as markers for malignancy enables the comparison of data under different illumination conditions and with different surface structures. Detection of protein aggregation, loss of its compact structure, increasing macromolecule rigidity, and loss of serum biological activity, in combination with the activation of serum phosphorescence may play a decisive role in non-invasive glioblastoma testing. 5. Conclusions 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. Further studies will be required to confirm that the methods used here will provide accurate tissue diagnostic information in vivo. Nomenclature of abbreviations CHP - Conditionally healthy people CT – Computed tomography GB - Glioblastoma IDH – isocitrate dehydrogenase Kyn - kynurenine MGB – Multiforme Glioblastome MGMT - O6-methylguanine-DNA methyltransferase MRI – Magnetic resonance imaging NADH – Nicotinamide adenine dinucleotide NIGT – Non-invasive glioblastoma testing Trp – L-tryptophan Declarations Availability of data and materials The datasets generated and/or analyzed during the current study are available from the corresponding author on reasonable request. Acknowledgements The authors would like to thank the patients and clinical staff who gave up their time to take part in this clinical study. Authors’ contributions IK, SG, YI and VB constructed the study design. IK, VB, KS and YV contributed to data interpretation and manuscript drafting. YI, YV and SG, contributed to the statistical analysis. IK, KS and SG prepared figures. KS, SG, and YI participated in the clinical investigation and contributed to the epidemiological data collection. SG and KS revised the manuscript. All authors read and approved the final manuscript. Consent for publication - Not applicable Funding sources This research was part of the research work of the Kharkiv National Medical University “Improvement and development of methods for diagnosis and surgical treatment of diseases and injuries of the abdominal cavity and chest, vessels of the upper and lower extremities using mini-invasive techniques in patients at high risk of postoperative complications” The number of state registration is 0116u00499. This research received no external funding. Ethical Approval and consent to participate This two-institution retrospective cohort study was handled in accordance with the Declaration of Helsinki. The use of registered data follows the General Data Protection Regulation of the European Union. The study and the use of data were consented to by the Ethics Committee of Kharkiv National Medical University, Ukraine ((Protocol No. 6, November 11, 2022). The number of state registration is 0116u00499. Conflicts of Interest All the authors declare no conflicts of interest. Informed consent process Informed consent was obtained from all participants included in the study. Information about authors: Yuriy O. Vinnyk , Ph.D., Doctor of Medical Sciences, Professor, Professor of the Department of Oncology, Radiation Therapy, Oncosurgery and Palliative Care, Kharkiv National Medical University, Kharkiv, Ukraine. email: [email protected] https://orcid.org/0000-0002-8995-2862 Igor A. Kryvoruchko , Ph.D., Doctor of Medical Sciences, Professor, Head of the Department of Surgery No. 2, Kharkiv National Medical University, Kharkiv, Ukraine. email: [email protected] ORCID: https://orcid.org/0000-0002-5525-701X Valeriy V. Boyko , Ph.D., Doctor of Medical Sciences, Professor, Head of the Department of Surgery No. 1, Kharkiv National Medical University, Kharkiv, Ukraine. email: [email protected] ORCID: http://orcid.org/0000-0002-3455-9705 Yulia V. Ivanova , Ph.D., Doctor of Medical Sciences, Professor, Professor of the Department of Surgery No. 1, Kharkiv National Medical University, Kharkiv, Ukraine. email: [email protected] https://orcid.org/0000-0003-4464-3035 Svetlana Gramatiuk , PhD, President Ukraine Association of Biobank, MSc Biobanking Lecturer, International Biobanking and Education, Medical University of Graz, Austria. email: [email protected] https://orcid.org/0000-0003-4238-7031 Karine Sargsyan, Ph.D ., Doctor of Medical Sciences, Professor, Head of the International Biobanking and Education, Medical University of Graz, Austria. email: [email protected] (KS) https://orcid.org/0000-0001-5853-4994 References Urbanska K., Sokolowska J., Szmidt M., Sysa P. Glioblastoma multiforme - an overview. 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J., Patterson M. S., Giles A., Wilson B. C., “Imaging of photodynamically generated singlet oxygen luminescence in vivo,” Photochem. Photobiol. 81(4), 941–943 (2005). Breitenbach T., Kuimova M. K., Gbur P., Hatz S., Schack N. B., Pedersen B. W., Lambert J. D. C., Poulsen L., Ogilby P. R., “Photosensitized production of singlet oxygen: spatially-resolved optical studies in single cells,” Photochem. Photobiol. Sci. 8(4), 442–452 (2009). Price M., Reiners J. J., Santiago A. M., Kessel D., “Monitoring singlet oxygen and hydroxyl radical formation with fluorescent probes during photodynamic therapy,” Photochem. Photobiol. 85(5), 1177–1181 (2009). Hu B., Zeng N., Liu Z., Ji Y., Xie W., Peng Q., Zhou Y., He Y., Ma H., “Two-dimensional singlet oxygen imaging with its near-infrared luminescence during photosensitization,” J. Biomed. Opt. 16(1), 016003 (2011). S. Lee, D. H. Vu, M. F. Hinds, S. J. Davis, J. A. O'Hara, and B. W. Pogue, “A singlet molecular oxygen imaging sensor for photodynamic therapy,” in Biomedical Optics, OSA Technical Digest (CD) (Optical Society of America, 2008), paper BTuC4. Lee S., Galbally-Kinney K. L., Murphy B. A., Davis S. J., Hasan T., Spring B., Tu Y., Pogue B. W., Isabelle M. E., O’Hara J. A., “In vivo PDT dosimetry: singlet oxygen emission and photosensitizer fluorescence,” Proc. SPIE 7551, 75510F (2010). Schmidt-Erfurth U., Hasan T., “Mechanisms of action of photodynamic therapy with verteporfin for the treatment of age-related macular degeneration,” Surv. Ophthalmol. 45(3), 195–214 (2000). Ayaru L., Wittmann J., Macrobert A. J., Novelli M., Bown S. G., Pereira S. P., “Photodynamic therapy using verteporfin photosensitization in the pancreas and surrounding tissues in the Syrian golden hamster,” Pancreatology 7(1), 20–27 (2007). O’Hara J., Samkoe K. S., Chen A., Hoopes P. J., Rizvi I., Hasan T., Pogue B. W., “Uptake of verteporfin by orthotopic xenograft pancreas models with different levels of aggression,” Proc. SPIE 7380, 73805F–, 73805F-7., 73805F-7 (2009). Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 27 Apr, 2023 Read the published version in Journal of Fluorescence → Version 1 posted Editorial decision: Major revision 18 Mar, 2023 Reviews received at journal 17 Mar, 2023 Reviewers agreed at journal 11 Mar, 2023 Reviewers invited by journal 10 Mar, 2023 Editor assigned by journal 10 Mar, 2023 Submission checks completed at journal 09 Mar, 2023 First submitted to journal 07 Mar, 2023 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-2665331","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":182274790,"identity":"121964af-cba0-429c-b641-61087d15b8d6","order_by":0,"name":"Yuriy O. Vinnyk","email":"","orcid":"","institution":"Kharkiv National Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Yuriy","middleName":"O.","lastName":"Vinnyk","suffix":""},{"id":182274791,"identity":"92019e34-265f-465a-a774-b764ee2d31d5","order_by":1,"name":"Igor A. Kryvoruchko","email":"","orcid":"","institution":"Kharkiv National Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Igor","middleName":"A.","lastName":"Kryvoruchko","suffix":""},{"id":182274792,"identity":"1317a2cd-c77b-4366-8a9e-3057221cf753","order_by":2,"name":"Valeriy V. Boyko","email":"","orcid":"","institution":"Institute General and Emergency Surgery named after V.T. Zaitcev of the National Academy of Medical Sciences of Ukraine","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Valeriy","middleName":"V.","lastName":"Boyko","suffix":""},{"id":182274793,"identity":"1919ca07-fef3-4973-b8ee-437a18ca75e7","order_by":3,"name":"Yulia V. Ivanova","email":"","orcid":"","institution":"Kharkiv National Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Yulia","middleName":"V.","lastName":"Ivanova","suffix":""},{"id":182274794,"identity":"0597db4f-3d10-471a-b158-e0b94a318d65","order_by":4,"name":"Svetlana Gramatiuk","email":"data:image/png;base64,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","orcid":"","institution":"Ukraine Association of Biobank","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Svetlana","middleName":"","lastName":"Gramatiuk","suffix":""},{"id":182274795,"identity":"536edd1f-db53-4a5b-bce8-27f00a3c8b76","order_by":5,"name":"Karine Sargsyan","email":"","orcid":"","institution":"Medical University of Graz","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Karine","middleName":"","lastName":"Sargsyan","suffix":""}],"badges":[],"createdAt":"2023-03-07 12:29:15","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-2665331/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-2665331/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s10895-023-03237-9","type":"published","date":"2023-04-27T20:38:46+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":34237880,"identity":"e4355d2a-819a-4d7b-94fe-f13d4ff67369","added_by":"auto","created_at":"2023-03-14 14:49:29","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":50696,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003ePhosphorescence spectra of D-L tryptophan powder for different excitation wavelengths cancerous brain tissues. Spectra are integrated from 0.5- to 7.5-ms delay concerning the excitation lamp pulse.\u003c/em\u003e\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-2665331/v1/c8ff06448d8f2177f701ee7e.jpg"},{"id":34237879,"identity":"01442134-e19b-41fb-a6ff-dfbff6fee55d","added_by":"auto","created_at":"2023-03-14 14:49:29","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":44830,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003ePhosphorescence from normal and malignant brain tissues. Signals were integrated for detector gate delays of 1 to 7 ms.\u003c/em\u003e\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-2665331/v1/36066d899c089dfe5550f62c.jpg"},{"id":44728817,"identity":"e5e5fd18-deff-4a2f-8ed5-352b8b657776","added_by":"auto","created_at":"2023-10-16 21:08:36","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":458503,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-2665331/v1/d6db2629-18cf-46a1-be16-819d345d5316.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Investigate the possibility of using phosphorescence in clinical oncology as an early prognostic test in detecting brain carcinogenesis","fulltext":[{"header":"1.\tIntroduction","content":"\u003cp\u003eGlioblastoma (GBM) is the most aggressive primary brain tumor, with an incidence of 2\u0026ndash;3 cases per 100,000 people [1]. The average survival in Ukraine of about 12-14 months is achieved with intensive multimodal treatment, but the number of such patients is minimal. Despite such intensive treatment, there is no cure, and the recurrence of Glioblastoma is inevitable [2].\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eApproaches to diagnosing GBM are rapidly evolving and are based on the recently revised WHO (2016) criteria for classifying central nervous system tumors [3]. Currently, histopathological examination of a tissue sample for suspected GBM is the gold standard for diagnosis. Now complemented by molecular diagnostics, the identification of O6-methylguanine-DNA methyltransferase (MGMT) methylation, isocitrate dehydrogenase (IDH) mutation, and 1p19q coding is currently the most valuable in daily clinical practice [4].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eSeveral studies have proven that cellular metabolism is the main regulator of tumor behavior [5]. To maintain a high proliferation rate, cancer cells undergo metabolic reprogramming, and tumors use amino acids to meet energy needs and biosynthesis in cells [6]. Tryptophan plays an essential role in malignant conversion and tumor progression. Trp degradation occurs via the kynurenine (Kyn) pathway, followed by the formation of metabolites [7]. Non-invasive glioblastoma testing (NIGT) combines non-invasive (non-surgical) techniques to represent the tumor and provides information about tumor drivers and the microenvironment, all factors that can/should be included in a treatment regimen. Among the large number of NIGTs using spectroscopic methods available for testing cell populations and the functional state of biological fluids, the phosphorescence method occupies a leading position. Phosphorescence spectroscopy of amino acids such as Tryptophan is a fast, easy-to-use, highly sensitive, and highly selective method that allows you to gain in-depth knowledge of the structural, functional, and metabolic changes in cells, organs, and systems that occur at the molecular level [8, 9, 10].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003ePhosphorescence is considered one of the NIGT 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 [11, 12\u0026ndash;16].\u003c/p\u003e\n\u003cp\u003eThis study aimed to conduct a feasibility study using phosphorescence in clinical oncology as an early prognostic test in detecting Glioblastoma.\u003c/p\u003e"},{"header":"2. Materials And Methods","content":"\u003cdiv id=\"Sec2\" class=\"Section2\"\u003e \u003ch2\u003e2.1. Patient selection and data collection\u003c/h2\u003e \u003cp\u003eThis 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). The respective institutional review boards of each participating institution have approved this study. Demographic and clinical data were collected, including age, gender, concurrent diseases, MRI, white blood cell count, and total neutrophil and lymphocyte count.\u003c/p\u003e \u003cp\u003e This retrospective cohort study was handled in accordance with the Declaration of Helsinki. This manuscript adheres to the applicable STROBE guideline. The use of registered data follows the General Data Protection Regulation of the European Union.\u003c/p\u003e \u003cp\u003e The patient\u0026rsquo;s written Informed consent was signed for each bio-object from the residual materials. The study and the use of data were consented to by the Ethics Committee of Kharkiv National Medical University, Ukraine (Protocol No. 6, November 11, 2022).\u003c/p\u003e \u003cp\u003eParticipants must meet one of the following: diagnosed with a single primary cancer that has not yet been treated. There is no evidence or treatment of any cancer for at least five years before enrollment.\u003c/p\u003e \u003cp\u003eKey Exclusion Criteria: a medical condition which, in the investigator's opinion, should preclude enrollment in the study. Known to be pregnant. Any therapy for cancer, including surgery, chemotherapy, immunotherapy, and/or radiation therapy, in the five years, preceding enrollment. Participated in or is currently participating in a clinical research study in which an experimental medication has been administered in the last 30 days. Participated in or is currently participating in another clinical study.\u003c/p\u003e \u003cp\u003eFor the control cohort: Any previous cancer diagnosis in the five years preceding enrollment, recurrence of the same primary cancer within any timeframe, or concurrent diagnosis of multiple primary cancers within any timeframe. The comparative group consisted of 486 conventionally healthy men and women aged from 25 to 65 years.\u003c/p\u003e \u003cp\u003eSerum proteins' phosphorescence state was studied and evaluated in 1039 patients with cancer brains (Glioblastoma), aged from 35 to 68 years. Clinical and histomorphological methods confirmed the diagnosis.\u003c/p\u003e \u003cp\u003eAmong 1039 patients were 764 (406 men, 358 women) patients with Glioblastoma (GB) and 275 (150 men and 125 women) with Multiforme Glioblastoma (MGB).\u003c/p\u003e \u003cp\u003eAfter detecting a brain tumor on CT or MRI, the neurosurgeon obtained tissue from the tumor for biopsy. We used tumor tissue analysis to assign tumor, severity and histological grade.\u003c/p\u003e \u003cp\u003eThe first stage (I) of the disease was diagnosed in 81 patients (these were cases of cystic brain disease), the second (II) in 163, the third (III) in 564, and the fourth stage (IV) was detected in 231 patients.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.2 Method of protein phosphorescence detection included two steps.\u003c/h2\u003e \u003cp\u003eDuring the first step, we obtain a blood sample (5.0 ml) from the ulnar vein and prepare serum by centrifugation using routine methods in medical practice. A study of luminol-dependent phosphorescence intensity in serum was carried out after its activation by the light source, according to the spectrofluorimeter method, as follows: 50 mcl of luminol-containing serum (10 mcl 3% luminol solution) was placed on a 5 x 45 mm quartz plate and placed in a thermostat. At a temperature of 30\u003csup\u003eo\u003c/sup\u003eC, serum drops were dried for 20 minutes 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. The spectral sensitivity of the FEP was located in the ultraviolet and visible ranges of daylight. Coating technology provides results in less than 0.5% d-spacing variation.\u003c/p\u003e \u003cp\u003eIn the second stage, we examined the tumor tissue after surgery. Fresh surgical specimens of cancerous brain tissues were acquired under Ukraine Association of Biobank approval from a tissue bank within three h of surgery. The standard tissue specimens were from the tumor's margins and were a mix of malignant and normal tissues. Samples were stored at 4\u0026deg;C and not subjected to additional processing before measuring the spectra. Specimens were cut into pieces ranging from \u0026sim;0.5 to \u0026sim;1 cm\u003csup\u003e2\u003c/sup\u003e and placed in a 1- x 1-cm quartz cuvette before mounting on the translation stage. Using the spectrograph setup, the spectra were acquired in a grid pattern with a 0.5 - to 1-mm spacing to cover most of the specimen. Each spectrum was integrated for 2s. Intensity ratio maps were generated from the individual spectra.\u003c/p\u003e \u003cp\u003eAll measurement procedures were automated, and the error was not more than 3% in all cases.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Statistical analysis\u003c/h2\u003e \u003cp\u003eThe summary statistics for continuous variables were calculated separately for each screening group by the number of subjects with the number of recorded observations (N), mean, standard deviation (SD), minimum (Min), first quartile (Q1), median, third quartile (Q3), and maximum (Max). Categorical data were displayed by the screening group using absolute frequencies and percentages.\u003c/p\u003e \u003cp\u003eFor continuous variables, a between-group comparison will be performed using the Student's t-test for independent samples or the Wilcoxon test, depending on the results of checking the normal distribution of data in groups using the Shapiro-Wilk test.\u003c/p\u003e \u003cp\u003eGroup comparisons will be made using Pearson's chi-squared test (χ2) for categorical variables. If, in any of the cells of the contingency table, the expected frequencies were less than 5, Fisher's exact test would be applied for comparison. Statistical processing of the obtained data was carried out using the software package Statistica v. 6.0 (Statsoft Inc.).\u003c/p\u003e \u003c/div\u003e"},{"header":"3. Results","content":"\u003cp\u003eThe main difference in the levels of serum phosphorescence between patients and conditionally healthy people was observed at the activation of a 404 nm spectral wave. So, the intensity of serum phosphorescence in cancer patients raised 3.6 and 3.8 times, respectively, in men and women. This wave of activation was appropriate for hemoglobin. It can be indicated the loss of compact structure and functional activity by this protein that may result from tissue hypoxia. Research of serum phosphorescence intensity in patients with brain cancer according to the type of tumor process revealed similar dynamics in all studying groups (Table 1).\u003c/p\u003e\n\u003cp\u003eTable\u0026nbsp;1\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eThe intensity of luminol-dependent serum phosphorescence in patients with brain cancer and conditionally healthy people depends on tumor type\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"621\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" valign=\"top\" width=\"14.814814814814815%\"\u003e\n \u003cp\u003eThe range of\u0026nbsp;activation (nm)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"65.37842190016103%\"\u003e\n \u003cp\u003ePhosphorescence intensity\u0026nbsp;( І\u003csup\u003e0\u003c/sup\u003eс), М\u0026plusmn;m\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"3\" valign=\"top\" width=\"19.806763285024154%\"\u003e\n \u003cp\u003e\u003cstrong\u003ec2/P value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"62.80788177339902%\"\u003e\n \u003cp\u003ePatients\u0026nbsp;with\u0026nbsp;Glioblastoma\u0026nbsp;(n=1039)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" width=\"37.19211822660098%\"\u003e\n \u003cp\u003eConditionally\u0026nbsp;healthy people\u0026nbsp;(n=486)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"48.23529411764706%\"\u003e\n \u003cp\u003eMGB\u0026nbsp;(n=275)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"51.76470588235294%\"\u003e\n \u003cp\u003eMG\u0026nbsp;(n=764)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"14.814814814814815%\"\u003e\n \u003cp\u003e297\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.806763285024154%\"\u003e\n \u003cp\u003e5997,8\u0026plusmn;364,5*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"21.256038647342994%\"\u003e\n \u003cp\u003e6731,4\u0026plusmn;356,7*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"24.315619967793882%\"\u003e\n \u003cp\u003e1.376/0.503\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.806763285024154%\"\u003e\n \u003cp\u003e1.376/0.503\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"14.814814814814815%\"\u003e\n \u003cp\u003e313\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.806763285024154%\"\u003e\n \u003cp\u003e437,6\u0026plusmn;65,7*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"21.256038647342994%\"\u003e\n \u003cp\u003e483,5\u0026plusmn;76,8*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"24.315619967793882%\"\u003e\n \u003cp\u003e12.469/0.014\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.806763285024154%\"\u003e\n \u003cp\u003e12.469/0.014\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"14.814814814814815%\"\u003e\n \u003cp\u003e334\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.806763285024154%\"\u003e\n \u003cp\u003e807,4\u0026plusmn;53,6*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"21.256038647342994%\"\u003e\n \u003cp\u003e796,3\u0026plusmn;68,4*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"24.315619967793882%\"\u003e\n \u003cp\u003e1.179/0.555\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.806763285024154%\"\u003e\n \u003cp\u003e1.179/0.555\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"14.814814814814815%\"\u003e\n \u003cp\u003e365\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.806763285024154%\"\u003e\n \u003cp\u003e3014,5\u0026plusmn;186,3*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"21.256038647342994%\"\u003e\n \u003cp\u003e2868,7\u0026plusmn;154,3*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"24.315619967793882%\"\u003e\n \u003cp\u003e8.752/0.013\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.806763285024154%\"\u003e\n \u003cp\u003e8.752/0.013\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"14.814814814814815%\"\u003e\n \u003cp\u003e404\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.806763285024154%\"\u003e\n \u003cp\u003e1894,3\u0026plusmn;145,8*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"21.256038647342994%\"\u003e\n \u003cp\u003e1812,4\u0026plusmn;133,6*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"24.315619967793882%\"\u003e\n \u003cp\u003e0.369/0.832\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.806763285024154%\"\u003e\n \u003cp\u003e0.369/0.832\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"14.814814814814815%\"\u003e\n \u003cp\u003e434\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.806763285024154%\"\u003e\n \u003cp\u003e1826,4\u0026plusmn;103,7*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"21.256038647342994%\"\u003e\n \u003cp\u003e1938,6\u0026plusmn;145,2*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"24.315619967793882%\"\u003e\n \u003cp\u003e0.927/0.629\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.806763285024154%\"\u003e\n \u003cp\u003e0.927/0.629\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eThe highest level of phosphorescence was observed at activation with a 297 nm spectral wave. In this case, serum phosphorescence raised at 86.8 % and 109.6% in patients with GB and MGB. The intensity of serum phosphorescence at activation with the spectral line of 404 nm was increased by 3.8 and 3.6 times, or by 271.4% and 258.6%, respectively, in patients with GB and MGB compared to a group of conditionally healthy people. The smaller difference in phosphorescence was observed at activation with spectra of 313 nm, 334 nm, and 365 nm. So, at 313 nm, phosphorescence intensity raised at 29.2 %, and 52.1%, at 334 nm \u0026ndash; at 26.7 %, and 21.7 %, at 365 nm \u0026ndash; at 67.2 %, and 62.9 % accordingly in patients with GB and MGB.\u003c/p\u003e\n\u003cp\u003eThe study of serum phosphorescence in GB patients revealed a high informativeness of indexes at activation with a spectral range of 297 nm, 404 nm, and 434 nm. It revealed a direct correlation between the degree of disease severity and the intensity of phosphorescence in GB patients\u0026apos; serum (Table 2).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eTable\u0026nbsp;2\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eThe intensity of luminol-dependent\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eserum phosphorescence\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003ein\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;patients depending on the stage of\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eGB and MGB\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" valign=\"top\" width=\"11.442006269592477%\"\u003e\n \u003cp\u003eThe range of\u0026nbsp;activation (nm)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"6\" valign=\"top\" width=\"88.55799373040752%\"\u003e\n \u003cp\u003ePhosphorescence intensity\u0026nbsp;( І\u003csup\u003e0\u003c/sup\u003eс) , М\u0026plusmn;m\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"4\" valign=\"top\" width=\"73.40425531914893%\"\u003e\n \u003cp\u003eStage\u0026nbsp;of GB and MGB,\u0026nbsp;n\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" width=\"17.02127659574468%\"\u003e\n \u003cp\u003eCHP\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;(n=486)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" width=\"9.574468085106384%\"\u003e\n \u003cp\u003e\u003cstrong\u003eP\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.93946731234867%\"\u003e\n \u003cp\u003eStage - I\u0026nbsp;\u003c/p\u003e\n \u003cp\u003en=\u0026nbsp;81\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.181598062953995%\"\u003e\n \u003cp\u003eStage \u0026ndash; II\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;n=163\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"24.93946731234867%\"\u003e\n \u003cp\u003eStage \u0026ndash; III\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;n=564\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"24.93946731234867%\"\u003e\n \u003cp\u003eStage \u0026ndash; IV\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;n=231\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"11.477987421383649%\"\u003e\n \u003cp\u003e297\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.19496855345912%\"\u003e\n \u003cp\u003e5706,3\u0026plusmn;127,5*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.352201257861637%\"\u003e\n \u003cp\u003e6157,8\u0026plusmn;104,5*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.19496855345912%\"\u003e\n \u003cp\u003e6508,3\u0026plusmn;118,6*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.19496855345912%\"\u003e\n \u003cp\u003e6837,4\u0026plusmn;162,7*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"15.09433962264151%\"\u003e\n \u003cp\u003e3210,7\u0026plusmn;137,5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.49056603773585%\"\u003e\n \u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"11.477987421383649%\"\u003e\n \u003cp\u003e313\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.19496855345912%\"\u003e\n \u003cp\u003e394,8\u0026plusmn;23,6*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.352201257861637%\"\u003e\n \u003cp\u003e415,6\u0026plusmn;32,7*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.19496855345912%\"\u003e\n \u003cp\u003e488,9\u0026plusmn;42,7*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.19496855345912%\"\u003e\n \u003cp\u003e520,6\u0026plusmn;43,8*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"15.09433962264151%\"\u003e\n \u003cp\u003e317,8\u0026plusmn;19,6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.49056603773585%\"\u003e\n \u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"11.477987421383649%\"\u003e\n \u003cp\u003e334\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.19496855345912%\"\u003e\n \u003cp\u003e762,5\u0026plusmn;33,8*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.352201257861637%\"\u003e\n \u003cp\u003e794,3\u0026plusmn;26,5*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.19496855345912%\"\u003e\n \u003cp\u003e825,6\u0026plusmn;53,2*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.19496855345912%\"\u003e\n \u003cp\u003e863,7\u0026plusmn;29,4*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"15.09433962264151%\"\u003e\n \u003cp\u003e653,9\u0026plusmn;39,6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.49056603773585%\"\u003e\n \u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"11.477987421383649%\"\u003e\n \u003cp\u003e365\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.19496855345912%\"\u003e\n \u003cp\u003e2796,8\u0026plusmn;75,3*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.352201257861637%\"\u003e\n \u003cp\u003e2837,6\u0026plusmn;83,4*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.19496855345912%\"\u003e\n \u003cp\u003e2925,8\u0026plusmn;77,4*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.19496855345912%\"\u003e\n \u003cp\u003e3097,6\u0026plusmn;105,8*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"15.09433962264151%\"\u003e\n \u003cp\u003e1760,5\u0026plusmn;59,2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.49056603773585%\"\u003e\n \u003cp\u003e\u0026lt;0.003\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"11.477987421383649%\"\u003e\n \u003cp\u003e404\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.19496855345912%\"\u003e\n \u003cp\u003e1754,6\u0026plusmn;56,3*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.352201257861637%\"\u003e\n \u003cp\u003e1814,3\u0026plusmn;76,8*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.19496855345912%\"\u003e\n \u003cp\u003e1897,2\u0026plusmn;83,6*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.19496855345912%\"\u003e\n \u003cp\u003e1972,4\u0026plusmn;63,5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"15.09433962264151%\"\u003e\n \u003cp\u003e505,3\u0026plusmn;39,4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.49056603773585%\"\u003e\n \u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"11.477987421383649%\"\u003e\n \u003cp\u003e434\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.19496855345912%\"\u003e\n \u003cp\u003e1710,3\u0026plusmn;64,5*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.352201257861637%\"\u003e\n \u003cp\u003e1794,8\u0026plusmn;73,6*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.19496855345912%\"\u003e\n \u003cp\u003e1923,8\u0026plusmn;105,4*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.19496855345912%\"\u003e\n \u003cp\u003e1986,7\u0026plusmn;84,3*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"15.09433962264151%\"\u003e\n \u003cp\u003e593,6\u0026plusmn;30,9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.49056603773585%\"\u003e\n \u003cp\u003e\u0026lt;0.002\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eThe correlation coefficients in all cases were between 0.87 (r = 0.87) and more. Levels of serum phosphorescence intensity in patients with GB and MGB in activation with 297 nm increased at 77.7 %, 91.7 %, 102.3 %, and 112.3 %, in 313 nm \u0026ndash; at 24.2 %, 30.7 %, 53.8 %, and 63.8 %, in 334 nm \u0026ndash; at 16.6 %, 21.4 %, 26.2 %, and 32.0 %, in 365 nm \u0026ndash; at 58.8 %, 61.2 %, 66.2 %, and 75.9 %, in 404 nm \u0026ndash; at 247.2 %, 259.5 %, 275.0 %, and 290.3 %, in 434 nm \u0026ndash; at 188.1%, 102, 3%, 224.1 %, and 234.6 %. In all cases, it was noted that phosphorescence intensity rose by more than 200 percent in activation with spectra of 404 nm and 434 nm. And also, a significant increase in phosphorescence was detected at activation with a 297 nm spectral wave.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eInactivation of serum with the lower ultraviolet (365 nm) phosphorescence increased by more than 60% compared to a group of conditionally healthy people. In all groups and at different spectral lines, a correlation between phosphorescence intensity and the stage of the tumor process took place.\u003c/p\u003e\n\u003cp\u003eThe phosphorescence spectra of D-L-trp powder excited at 282, 300, 380, and 400 nm and acquired with the CD-Scan cancerous brain tissues are shown in Figure 1. Each spectrum shown in the figure is the sum of seven ranges, obtained with delays of 1 to 7 ms in 1-ms steps.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe gate width was 1 ms. Thus, each range shown is the integrated intensity from t = 0.5 to 7.5 ms. Although it is well known that the trp excitation maximum is at 282 nm and that trp does not have significant absorption at wavelengths longer than 300 nm, the phosphorescence intensity with 282 nm excitation was weaker than with 400 nm excitation. The phosphorescence was blue-shifted for the longer wavelength excitations (380 and 400 nm) compared to the shorter wavelength excitations (282 and 300 nm). For 380 nm and 400 nm excitation, trp exhibits two phosphorescence peaks at 480 nm and 525 nm. For excitation at 282 nm and 300 nm, only the phosphorescence peak at 525 nm was observed. The mechanism responsible for the shift in phosphorescence and the reason for the more intense at 400 nm excitation is unknown. It may be due to dimers or trimers present in the trp powder.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003ePhosphorescence from ex vivo human brain tissues\u003c/em\u003e. Although the phosphorescence intensity in trp powder was greater for excitation at 400 nm than for excitation at 300 nm, tissues contain several fluorophores (NADH and flavins) whose emission wavelengths can overlap with the trp. Therefore, the brain tissue specimens were excited at 300 nm to reduce the contribution from these fluorophores. Figure 2 shows the phosphorescence from ex vivo human normal and malignant brain tissues acquired with the CD-Scan. The plots shown in the figure are each the integration of seven spectra obtained with gate delays of 1 to 7 ms with 1-ms intervals. The normal tissues exhibited phosphorescence emission from 440 to 500 nm. Fluorescence can also be observed at 350 nm \u0026mdash; most likely due to a long \u0026ldquo;tail\u0026rdquo; on the lamp emission. The normal tissues showed more excellent fluorescence and phosphorescence than the malignant tissues, which had almost no detectable phosphorescence.\u003c/p\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eOptical biopsy\u0026mdash;using native tissue fluorescence\u0026mdash;for detecting cancer has been an area of investigation for over two decades. The bases behind the operation of optical biopsy are that the onset of carcinogenesis results in structural changes (thickening of the mucosa layer, increased vascularity) and molecular changes (increased nucleic acids, alterations in protein structure, increased cell metabolism) that modify the spectroscopic properties of tissue and, thereby, create unique optical signatures that can be used to detect malignant and premalignant tissues. In the UV and blue spectral regions, the primary native tissue fluorophores are Tryptophan (trp), collagen, elastin, reduced nicotinamide adenine dinucleotide (NADH), and flavins [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. Optical biopsy has been demonstrated to be an accurate, real-time tool for distinguishing normal tissues from malignant and premalignant tissues. Prior investigations have demonstrated that several ratio-based algorithms can distinguish malignant tissue from normal tissue with high sensitivity and specificity. The wavelength combinations used in those ratios isolated the contributions from different pairs of tissue fluorophores, one of which was trp. The ratio of 340 nm to 440 nm emission (with 300 nm excitation) is useful for identifying malignant tissues from many different organ sites [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAccording to the findings, the appearance of many molecules in the triplet state in the long-wavelength spectral region of activation may indicate a dissociation of oxidative phosphorylation. The tissue respiration, inhibition of bioenergy processes, which are accompanied by ineffective use of energy and scattering it in the form of heat, and reduced ATP production are symptoms of mitochondrial pathology [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e, \u003cspan additionalcitationids=\"CR23\" citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. The presence of high energy levels in activated electronic states due to the appearance of unpaired electrons in the active molecule indicates a change in the reaction ability and conformational properties of proteins, nucleic acids, and other biologically active molecules (enzymes, hormones). It is known that ultraviolet spectrum photons are absorbed mainly by aromatic amino acids (tyrosine - at 280 nm, Tryptophan - at 220 nm), proteins (at 280\u0026ndash;300 nm), nucleic acids, and nucleotides (at 260 nm) which may be present in the serum. An analysis of the results reveals that profound metabolic, structural, and conformational changes of large polymer molecules and their monomeric components occurred in patients with brain cancer. Increased serum phosphorescence intensity in the long-wave region (404\u0026ndash;434 nm) may indicate an increase in non-erythrocytic hemoglobin levels and violations in its compact structure, conformational properties, and hemin content (404 nm) as a result. The development of free radical-membrane pathology in patients with brain cancer underpins the formation of hypochromic anemia in carcinogenesis.\u003c/p\u003e \u003cp\u003eThe study of serum phosphorescence intensity in patients with MGB and MG detected violations of the structural and conformational properties of biologically important macromolecules (proteins, nucleic acids, hemoglobin, and other glycoproteins). The presence of many electron-activated molecules, which can stimulate free radical processes and disconnect oxidative phosphorylation, causes energy deficiency, tissue hypoxia, and membrane molecular pathology, all of which are pathogenic factors in brain carcinogenesis [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eLevels of luminol-dependent serum phosphorescence intensity at activation with 297, 404, and 434 nm can be used to diagnose disease severity and the state of bioenergy processes and plan surgery and pathogenetic therapy. Such spectral monochromatic activation waves as 297, 404, and 434 nm are the most informative. They indicate a disturbance in serum proteins' compact structure and conformational properties and decreased biological activity. High levels of phosphorescence intensity in the first stage of carcinogenesis show changes in the compact design and biological activity of proteins for quite a long period, which precedes the development of the tumor process and is a prognostic factor in the development of oncopathology and its early diagnostic criteria [\u003cspan additionalcitationids=\"CR28\" citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eProlonged activation of oxidative processes leads to changes in protein conformation and biological activity observed in patients with brain cancer. The appearance in the long-wavelength spectral region of activation (404 and 434 nm) and the increased number of molecules in the triplet state may also indicate disconnection of oxidative phosphorylation and tissue respiration, which is always accompanied by heat energy scattering and mitochondrial pathology development, which are pathogenetic factors in carcinogenesis. Increased serum phosphorescence intensity in the ultraviolet region of 297 nm indicates the presence of high levels of triplet-activated states. It shows the change in proteins and nucleic acids' compact structures. The increase in serum phosphorescence in brain cancer patients in the long-wave spectral region of activation (404 nm) reflects an increase in free hemins (a nonprotein component of hemoglobin), confirming the protein's loss of compact structure and biological activity.\u003c/p\u003e \u003cp\u003eInformative monochromatic spectral waves of activation in evaluating serum compact structure and biological activity were 297, 404, and 434 nm. Physiological indexes of luminol-dependent serum phosphorescence intensity at activation with 297 nm range from 3000 to 3500 imp/sec. Increasing luminol-dependent serum phosphorescence intensity at activation with 404 nm from 3500 to 5000 and with 434 nm from 400 to 1700 imp/s indicates a conformational change in the structure and biological activity of serum proteins; this can be a significant prognostic indicator in the early diagnosis of the precancerous metabolic state that leads to carcinogenesis and stage-dependent pathological processes. A further rise in luminol-dependent serum phosphorescence intensity from 5000 to 7000 imp/s at 297 nm and from 1700 to 2000 imp/s at 404 and 434 nm indicates an increase in the severity of the disease.\u003c/p\u003e \u003cp\u003eThis newly developed optical technique for cancer detection, based on phosphorescence and fluorescence spectroscopy, is fast, minimally invasive, and non-destructive. This technique may apply to in vivo or ex vivo tissue analysis. The 345 ∕ 500 ratio, with excitation at 300 nm, provides an excellent fingerprint for cancer detection in ex vivo tissues. A 345 ∕ 500 ratio of 0.5 to 12 corresponds to normal brain tissue, and a ratio higher than 12 indicates cancerous brain tissue. Using ratios as markers for malignancy enables the comparison of data under different illumination conditions and with different surface structures.\u003c/p\u003e \u003cp\u003eDetection of protein aggregation, loss of its compact structure, increasing macromolecule rigidity, and loss of serum biological activity, in combination with the activation of serum phosphorescence may play a decisive role in non-invasive glioblastoma testing.\u003c/p\u003e"},{"header":"5. Conclusions","content":"\u003cp\u003eConsidering 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.\u003c/p\u003e \u003cp\u003eFurther studies will be required to confirm that the methods used here will provide accurate tissue diagnostic information in vivo.\u003c/p\u003e"},{"header":"Nomenclature of abbreviations","content":"\u003cp\u003eCHP - Conditionally healthy people\u003c/p\u003e\n\u003cp\u003eCT \u0026ndash; Computed tomography\u003c/p\u003e\n\u003cp\u003eGB - Glioblastoma\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eIDH \u0026ndash; isocitrate dehydrogenase\u003c/p\u003e\n\u003cp\u003eKyn - kynurenine\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eMGB \u0026ndash; Multiforme Glioblastome\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eMGMT - \u0026nbsp;O6-methylguanine-DNA methyltransferase\u003c/p\u003e\n\u003cp\u003eMRI \u0026ndash; Magnetic resonance imaging\u003c/p\u003e\n\u003cp\u003eNADH \u0026ndash; Nicotinamide adenine dinucleotide\u003c/p\u003e\n\u003cp\u003eNIGT \u0026ndash; Non-invasive glioblastoma testing\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eTrp \u0026ndash; L-tryptophan\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets generated and/or analyzed during the current study are available from the corresponding author on reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors would like to thank the patients and clinical staff who gave up their time to take part in this clinical study. \u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026rsquo; contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIK, SG, YI and VB constructed the study design. IK, VB, KS and YV contributed to data interpretation and manuscript drafting. YI, YV and SG, contributed to the statistical analysis. IK, KS and SG prepared figures.\u003c/p\u003e\n\u003cp\u003eKS, SG, and YI participated in the clinical investigation and contributed to the epidemiological data collection. SG and KS revised the manuscript.\u003c/p\u003e\n\u003cp\u003eAll authors read and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e- Not applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding sources\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research was part of the research work of the Kharkiv National Medical University \u0026ldquo;Improvement and development of methods for diagnosis and surgical treatment of diseases and injuries of the abdominal cavity and chest, vessels of the upper and lower extremities using mini-invasive techniques in patients at high risk of postoperative complications\u0026rdquo; The number of state registration is 0116u00499. This research received no external funding.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical Approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis two-institution retrospective cohort study was handled in accordance with the Declaration of Helsinki. The use of registered data follows the General Data Protection Regulation of the European Union. The study and the use of data were consented to by the Ethics Committee of Kharkiv National Medical University, Ukraine ((Protocol No. 6, November 11, 2022). The number of state registration is 0116u00499.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflicts of Interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll the authors declare no conflicts of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInformed consent process\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eInformed consent was obtained from all participants included in the study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInformation about authors:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eYuriy O. Vinnyk\u003c/em\u003e\u003c/strong\u003e, Ph.D., Doctor of Medical Sciences, Professor, Professor of the Department of Oncology, Radiation Therapy, Oncosurgery and Palliative Care, Kharkiv National Medical University, Kharkiv, Ukraine. \u003c/p\u003e\n\u003cp\u003eemail: [email protected]\u003c/p\u003e\n\u003cp\u003ehttps://orcid.org/0000-0002-8995-2862\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eIgor A. Kryvoruchko\u003c/em\u003e\u003c/strong\u003e, Ph.D., Doctor of Medical Sciences, Professor, Head of the Department of Surgery No. 2, Kharkiv National Medical University, Kharkiv, Ukraine.\u003c/p\u003e\n\u003cp\u003eemail: [email protected]\u003c/p\u003e\n\u003cp\u003eORCID: https://orcid.org/0000-0002-5525-701X\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eValeriy V. Boyko\u003c/em\u003e\u003c/strong\u003e, Ph.D., Doctor of Medical Sciences, Professor, Head of the Department of Surgery No. 1, Kharkiv National Medical University, Kharkiv, Ukraine.\u003c/p\u003e\n\u003cp\u003eemail: [email protected]\u003c/p\u003e\n\u003cp\u003eORCID: http://orcid.org/0000-0002-3455-9705\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eYulia V. Ivanova\u003c/em\u003e\u003c/strong\u003e, Ph.D., Doctor of Medical Sciences, Professor, Professor of the Department of Surgery No. 1, Kharkiv National Medical University, Kharkiv, Ukraine.\u003c/p\u003e\n\u003cp\u003eemail: [email protected] \u003c/p\u003e\n\u003cp\u003ehttps://orcid.org/0000-0003-4464-3035\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eSvetlana Gramatiuk\u003c/em\u003e\u003c/strong\u003e, PhD, President Ukraine Association of Biobank, MSc Biobanking Lecturer, International Biobanking and Education, Medical University of Graz, Austria.\u003c/p\u003e\n\u003cp\u003eemail: [email protected]\u003c/p\u003e\n\u003cp\u003ehttps://orcid.org/0000-0003-4238-7031\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eKarine Sargsyan, Ph.D\u003c/em\u003e\u003c/strong\u003e., Doctor of Medical Sciences, Professor, Head of the International Biobanking and Education, Medical University of Graz, Austria.\u003c/p\u003e\n\u003cp\u003eemail: [email protected] (KS)\u003c/p\u003e\n\u003cp\u003ehttps://orcid.org/0000-0001-5853-4994\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eUrbanska K., Sokolowska J., Szmidt M., Sysa P. Glioblastoma multiforme - an overview. Contemporary oncology. 2014;18(5):307\u0026ndash;312. doi: 10.5114/wo.2014.40559.\u003c/li\u003e\n\u003cli\u003eStupp R., Mason W. P., van den Bent M. J., et al. Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. The New England Journal of Medicine. 2005;352(10):987\u0026ndash;996. doi: 10.1056/NEJMoa043330.\u003c/li\u003e\n\u003cli\u003eLouis D. N., Perry A., Reifenberger G., et al. The 2016 World Health Organization classification of tumors of the central nervous system: a summary. Acta Neuropathologica. 2016;131(6):803\u0026ndash;820. doi: 10.1007/s00401-016-1545-1.\u003c/li\u003e\n\u003cli\u003eHegi M. E., Diserens A. C., Gorlia T., et al. MGMT gene silencing and benefit from temozolomide in Glioblastoma. The New England Journal of Medicine. 2005;352(10):997\u0026ndash;1003. doi: 10.1056/NEJMoa043331.\u003c/li\u003e\n\u003cli\u003ePavlova NN, Thompson CB. The emerging hallmarks of Cancer metabolism. Cell Metab. 2016;23(1):27\u0026ndash;47. https://doi.org/10.1016/j.cmet.2015.12.006.\u003c/li\u003e\n\u003cli\u003eLieu EL, Nguyen T, Rhyne S, et al. Amino acids in cancer. Exp Mol Med. 2020;52(1):15\u0026ndash;30. https://doi.org/10.1038/s12276-020-0375-3.\u003c/li\u003e\n\u003cli\u003eOpitz CA, Somarribas Patterson LF, Mohapatra SR, et al. The therapeutic potential of targeting tryptophan catabolism in cancer. Br J Cancer. 2020;122(1):30\u0026ndash;44. https://doi.org/10.1038/s41416-019-0664-6.\u003c/li\u003e\n\u003cli\u003eKessel D., Luo Y., Mathieu P., Reiners J. J., Jr, \u0026ldquo;Determinants of the apoptotic response to lysosomal photodamage,\u0026rdquo; Photochem. Photobiol. 71(2), 196\u0026ndash;200 (2000). \u003c/li\u003e\n\u003cli\u003eFingar V. H., Wieman T. J., Wiehle S. A., Cerrito P. B., \u0026ldquo;The role of microvascular damage in photodynamic therapy: the effect of treatment on vessel constriction, permeability, and leukocyte adhesion,\u0026rdquo; Cancer Res. 52(18), 4914\u0026ndash;4921 (1992). \u003c/li\u003e\n\u003cli\u003eNiedre M. J., Patterson M. S., Wilson B. C., \u0026ldquo;Direct near-infrared luminescence detection of singlet oxygen generated by photodynamic therapy in cells in vitro and tissues in vivo,\u0026rdquo; Photochem. Photobiol. 75(4), 382\u0026ndash;391 (2002). \u003c/li\u003e\n\u003cli\u003eWei Y. C., Zhou J., Xing D., Chen Q., \u0026ldquo;In vivo monitoring of singlet oxygen using delayed chemiluminescence during photodynamic therapy,\u0026rdquo; J. Biomed. Opt. 12(1), 014002 (2007).\u003c/li\u003e\n\u003cli\u003eLee S., Galbally-Kinney K. L., Murphy B. A., Davis S. J., Hasan T., Spring B., Tu Y., Pogue B. W., Isabelle M. E., O\u0026rsquo;Hara J. A., \u0026ldquo;In vivo PDT dosimetry: singlet oxygen emission and photosensitizer fluorescence,\u0026rdquo; Proc. SPIE 7551, 75510F (2010). \u003c/li\u003e\n\u003cli\u003eSchmidt-Erfurth U., Hasan T., \u0026ldquo;Mechanisms of action of photodynamic therapy with verteporfin for the treatment of age-related macular degeneration,\u0026rdquo; Surv. Ophthalmol. 45(3), 195\u0026ndash;214 (2000). \u003c/li\u003e\n\u003cli\u003eAyaru L., Wittmann J., Macrobert A. J., Novelli M., Bown S. G., Pereira S. P., \u0026ldquo;Photodynamic therapy using verteporfin photosensitization in the pancreas and surrounding tissues in the Syrian golden hamster,\u0026rdquo; Pancreatology 7(1), 20\u0026ndash;27 (2007). \u003c/li\u003e\n\u003cli\u003eO\u0026rsquo;Hara J., Samkoe K. S., Chen A., Hoopes P. J., Rizvi I., Hasan T., Pogue B. W., \u0026ldquo;Uptake of verteporfin by orthotopic xenograft pancreas models with different levels of aggression,\u0026rdquo; Proc. SPIE 7380, 73805F\u0026ndash;, 73805F-7., 73805F-7 (2009).\u003c/li\u003e\n\u003cli\u003eLee S., Zhu L., Minhaj A. M., Hinds M. F., Vu D. H., Rosen D. I., Davis S. J., Hasan T., \u0026ldquo;Pulsed diode laser-based monitor for singlet molecular oxygen,\u0026rdquo; J. Biomed. Opt. 13(3), 034010 (2008). \u003c/li\u003e\n\u003cli\u003eLee S., Vu D. H., Hinds M. F., Davis S. J., Liang A., Hasan T., \u0026ldquo;Pulsed diode laser-based singlet oxygen monitor for photodynamic therapy: in vivo studies of tumor-laden rats,\u0026rdquo; J. Biomed. Opt. 13(6), 064035 (2008). doi: 10.1117/1.3042265. \u003c/li\u003e\n\u003cli\u003eLaubach H. J., Chang S. K., Lee S., Rizvi I., Zurakowski D., Davis S. J., Taylor C. R., Hasan T., \u0026ldquo;In-vivo singlet oxygen dosimetry of clinical 5-aminolevulinic acid photodynamic therapy,\u0026rdquo; J. Biomed. Opt. 13(5), 050504 (2008). \u003c/li\u003e\n\u003cli\u003eAndersen L. K., Gao Z., Ogilby P. R., Poulsen L., Zebger I., \u0026ldquo;A Singlet oxygen image with 2.5 \u0026mu;m resolution,\u0026rdquo; J. Phys. Chem. A 106(37), 8488\u0026ndash;8490 (2002). \u003c/li\u003e\n\u003cli\u003eZebger I., Snyder J. W., Andersen L. K., Poulsen L., Gao Z., Lambert J. D. C., Kristiansen U., Ogilby P. R., \u0026ldquo;Direct optical detection of singlet oxygen from a single cell,\u0026rdquo; Photochem. Photobiol. 79(4), 319\u0026ndash;322 (2004). \u003c/li\u003e\n\u003cli\u003eNiedre M. J., Patterson M. S., Giles A., Wilson B. C., \u0026ldquo;Imaging of photodynamically generated singlet oxygen luminescence in vivo,\u0026rdquo; Photochem. Photobiol. 81(4), 941\u0026ndash;943 (2005). \u003c/li\u003e\n\u003cli\u003eBreitenbach T., Kuimova M. K., Gbur P., Hatz S., Schack N. B., Pedersen B. W., Lambert J. D. C., Poulsen L., Ogilby P. R., \u0026ldquo;Photosensitized production of singlet oxygen: spatially-resolved optical studies in single cells,\u0026rdquo; Photochem. Photobiol. Sci. 8(4), 442\u0026ndash;452 (2009).\u003c/li\u003e\n\u003cli\u003ePrice M., Reiners J. J., Santiago A. M., Kessel D., \u0026ldquo;Monitoring singlet oxygen and hydroxyl radical formation with fluorescent probes during photodynamic therapy,\u0026rdquo; Photochem. Photobiol. 85(5), 1177\u0026ndash;1181 (2009). \u003c/li\u003e\n\u003cli\u003eHu B., Zeng N., Liu Z., Ji Y., Xie W., Peng Q., Zhou Y., He Y., Ma H., \u0026ldquo;Two-dimensional singlet oxygen imaging with its near-infrared luminescence during photosensitization,\u0026rdquo; J. Biomed. Opt. 16(1), 016003 (2011). \u003c/li\u003e\n\u003cli\u003eS. Lee, D. H. Vu, M. F. Hinds, S. J. Davis, J. A. O\u0026apos;Hara, and B. W. Pogue, \u0026ldquo;A singlet molecular oxygen imaging sensor for photodynamic therapy,\u0026rdquo; in Biomedical Optics, OSA Technical Digest (CD) (Optical Society of America, 2008), paper BTuC4.\u003c/li\u003e\n\u003cli\u003eLee S., Galbally-Kinney K. L., Murphy B. A., Davis S. J., Hasan T., Spring B., Tu Y., Pogue B. W., Isabelle M. E., O\u0026rsquo;Hara J. A., \u0026ldquo;In vivo PDT dosimetry: singlet oxygen emission and photosensitizer fluorescence,\u0026rdquo; Proc. SPIE 7551, 75510F (2010). \u003c/li\u003e\n\u003cli\u003eSchmidt-Erfurth U., Hasan T., \u0026ldquo;Mechanisms of action of photodynamic therapy with verteporfin for the treatment of age-related macular degeneration,\u0026rdquo; Surv. Ophthalmol. 45(3), 195\u0026ndash;214 (2000). \u003c/li\u003e\n\u003cli\u003eAyaru L., Wittmann J., Macrobert A. J., Novelli M., Bown S. G., Pereira S. P., \u0026ldquo;Photodynamic therapy using verteporfin photosensitization in the pancreas and surrounding tissues in the Syrian golden hamster,\u0026rdquo; Pancreatology 7(1), 20\u0026ndash;27 (2007). \u003c/li\u003e\n\u003cli\u003eO\u0026rsquo;Hara J., Samkoe K. S., Chen A., Hoopes P. J., Rizvi I., Hasan T., Pogue B. W., \u0026ldquo;Uptake of verteporfin by orthotopic xenograft pancreas models with different levels of aggression,\u0026rdquo; Proc. SPIE 7380, 73805F\u0026ndash;, 73805F-7., 73805F-7 (2009). \u003c/li\u003e\n\u003c/ol\u003e\n"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"journal-of-fluorescence","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"jofl","sideBox":"Learn more about [Journal of Fluorescence](https://www.springer.com/journal/10895)","snPcode":"10895","submissionUrl":"https://submission.nature.com/new-submission/10895/3","title":"Journal of Fluorescence","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"phosphorescence, non-invasive glioblastoma testing, L-tryptophan, cancerous brain tissues.","lastPublishedDoi":"10.21203/rs.3.rs-2665331/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-2665331/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003ePhosphorescence 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.\u003c/p\u003e\n\u003cp\u003eThis study aimed to conduct a feasibility study using phosphorescence in clinical oncology as an early prognostic test in detecting Glioblastoma.\u003c/p\u003e\n\u003cp\u003eMaterials and Methods. 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\u003csup\u003eo\u003c/sup\u003eC, serum drops were dried for 20 minutes 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.\u003c/p\u003e\n\u003cp\u003eResults and conclusion. 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.\u003c/p\u003e","manuscriptTitle":"Investigate the possibility of using phosphorescence in clinical oncology as an early prognostic test in detecting brain carcinogenesis","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2023-03-14 14:49:24","doi":"10.21203/rs.3.rs-2665331/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Major revision","date":"2023-03-18T11:39:31+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2023-03-17T13:34:49+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"0c602d8b-65a3-4524-a0f9-8794b16a5015","date":"2023-03-11T05:26:05+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2023-03-10T21:11:56+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2023-03-10T13:43:18+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2023-03-10T01:29:02+00:00","index":"","fulltext":""},{"type":"submitted","content":"Journal of Fluorescence","date":"2023-03-07T12:15:36+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"journal-of-fluorescence","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"jofl","sideBox":"Learn more about [Journal of Fluorescence](https://www.springer.com/journal/10895)","snPcode":"10895","submissionUrl":"https://submission.nature.com/new-submission/10895/3","title":"Journal of Fluorescence","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"38e6d0ce-350b-4a8c-bb7b-c9bfb63dba8d","owner":[],"postedDate":"March 14th, 2023","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2023-10-16T20:56:18+00:00","versionOfRecord":{"articleIdentity":"rs-2665331","link":"https://doi.org/10.1007/s10895-023-03237-9","journal":{"identity":"journal-of-fluorescence","isVorOnly":false,"title":"Journal of Fluorescence"},"publishedOn":"2023-04-27 20:38:46","publishedOnDateReadable":"April 27th, 2023"},"versionCreatedAt":"2023-03-14 14:49:24","video":"","vorDoi":"10.1007/s10895-023-03237-9","vorDoiUrl":"https://doi.org/10.1007/s10895-023-03237-9","workflowStages":[]},"version":"v1","identity":"rs-2665331","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-2665331","identity":"rs-2665331","version":["v1"]},"buildId":"7rjqhiLT3MXkJMwkYKINL","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}