Distinct Lipid Phenotype of Cancer-Associated Fibroblasts (CAFs) Isolated From Overweight/Obese Endometrial Cancer Patients as Assessed Using Raman Spectroscopy.

Obesity is strongly linked with increased risk and poorer prognosis of endometrial cancer (EC). Cancer-associated fibroblasts (CAFs) are activated fibroblasts that form a large component of the tumor microenvironment and undergo metabolic reprogramming to provide critical metabolites for tumor growth. However, it is still unknown how obesity, characterized by a surplus of free fatty acids drives the modifications of CAFs lipid metabolism which may provide the mechanistic link between obesity and EC progression. The present study aims to evaluate the utility of Raman spectroscopy, an emerging nondestructive analytical tool to detect signature changes in lipid metabolites of CAFs from EC patients with varying body mass index. We established primary cultures of fibroblasts from human EC tissues, and CAFs of overweight/obese and nonobese women using antibody-conjugated magnetic beads isolation. These homogeneous fibroblast cultures expressed fibroblast markers, including α-smooth muscle actin and vimentin. Analysis was made in the Raman spectra region best associated with cancer progression biochemical changes in lipids (600-1800 cm-1 and 2800-3200 cm-1). Direct band analysis and ratiometric analysis were conducted to extract information from the Raman spectrum. Present results demonstrated minor shifts in the CH2 symmetric stretch of lipids at 2879 cm-1 and CH3 asymmetric stretching from protein at 2932 cm-1 in the overweight/obese CAFS compared to nonobese CAFs, indicating increased lipid content and a higher degree of lipid saturation. Principal component analysis showed that CAFs from overweight/obese and nonobese EC patients can be clearly distinguished indicating the capability of Raman spectroscopy to detect changes in biochemical components. Our results suggest Raman spectroscopy supported by chemometric analysis is a reliable technique for characterizing metabolic changes in clinical samples, providing an insight into obesity-driven alteration in CAFs, a critical stromal component during EC tumorigenesis.

Thermoluminescent characterization and defect studies of graphite-rich media under high dose neutron exposure.

Thermoluminescence (TL) materials have a broad variety of uses in various fields, such as clinical research, individual dosimetry, and environmental dosimetry, amongst others. However, the use of individual neutron dosimetry has been developing more aggressively lately. In this regard, present study establishes a relationship between the neutron dosage and the optical property changes of graphite-rich materials caused by high doses of neutron radiation. This has been done with the intention of developing a novel, graphite-based radiation dosimeter. Herein, the TL yield of commercially graphite-rich materials (i.e. graphite sheet, 2B and HB grade pencils) irradiated by neutron radiation with doses ranging from 250 Gy to 1500 Gy has been investigated. The samples were bombarded with thermal neutrons as well as a negligible amount of gamma rays, from the nuclear reactor TRIGA-II installed at the Bangladesh Atomic Energy Commission. The shape of the glow curves was observed to be independent of the given dosage, with the predominant TL dosimetric peak maintained within the region of 163 °C-168 °C for each sample. By studying the glow curves of the irradiated samples, some of the most well theoretical models and techniques were used to compute the kinetic parameters such as the order of kinetics (b), activation energy (E) or trap depth, frequency factor (s) or escape probability, and trap lifetime (τ). All of the samples were found to have a good linear response over the whole dosage range, with 2B grade of polymer pencil lead graphite (PPLGs) demonstrating a higher level of sensitivity than both HB grade and graphite sheet (GS) samples. Additionally, the level of sensitivity shown by each of them is highest at the lowest dosage that was given, and it decreases as the dose increases. Importantly, the phenomenon of dose-dependent structural modifications and internal annealing of defects has been observed by assessing the area of deconvoluted micro-Raman spectra of graphite-rich materials in high-frequency areas. This trend is consistent with the cyclical pattern reported in the intensity ratio of defect and graphite modes in previously investigated carbon-rich media. Such recurrent occurrences suggest the idea of employing Raman microspectroscopy as a radiation damage study tool for carbonaceous materials. The excellent responses of the key TL properties of the 2B grade pencil demonstrate its usefulness as a passive radiation dosimeter. As a consequence, the findings suggest that graphite-rich materials have the potential to be useful as a low-cost passive radiation dosimeter, with applications in radiotherapy and manufacturing.

Dosimetric characteristics of Gd-doped silica glass subjected to neutron and gamma irradiations.

The dosimetric characteristics of newly developed gadolinium (Gd) glass dosimeter produced via sol-gel method are reported. Irradiation were made using a 750 kW neutron flux thermal power and 1.25 MeV 60Co gamma rays with entrance doses from 2 to 10 Gy. Investigation has been done on various Gd dopant concentrations, ranging from 1 to 10 mol%. The Gd-doped silica glass have been characterised for thermoluminescence (TL) dose response, sensitivity, linearity index, glow curve and kinetic parameter analysis. For particular dopant concentration obtained in 6 mol% Gd, the least squares fit shows the change in TL yield, correlation coefficient (r2) of better than 0.980 (at 95% confidence level), with neutron and gamma exposure to be 8 and 4 times greater than that of 1 mol% Gd, respectively. Broad peaks in the absence of any sharp peak observed in the glow curve confirms the amorphous nature of the prepared glass. A glow curve of Gd-doped SiO2 sample is observed with a single prominent peak (Tm) within 200-250 °C (peak shifting appears with respect to the increment of dopant concentration) and 350 °C (for all respective Gd dopants) for neutron and gamma irradiations, respectively. Deconvolution shows the glow curves of the Gd-doped SiO2 glass to be formed of seven and five overlapping peaks, with figures of merit below 2% (FOM) of between 1.38-1.79 and 1.30-1.97 for the particular neutron and gamma irradiations, respectively. Through use of Glowfit deconvolution software, the key trapping parameters of activation energy, E and frequency factor, s-1 were calculated for the Gd-doped SiO2 glass. The mechanism of TL yield with the gradual increase in Gd concentrations and doses is explained upon the incorporation of Gd and radiation damage that change the structure of the electron traps in the glass matrix. These early results indicate that selectively screened Gd-SiO2 glass can be developed into a promising TL system towards dosimetric applications.

Low-cost commercial graphite-rich pencils subjected to electron irradiation for passive radiation dosimetry.

Various thicknesses of 2B grade polymer pencil lead graphite (PPLG) were used in the present study, which focussed on the alteration in crystalline lattice and the structural defect caused by the electron irradiation dosage ranging from 0.5 to 20 Gy delivered by an Elekta HD Linac. The fundamental trap parameters i.e. kinetics order (b), activation energy (E), and frequency factor (s) of the PPLG samples have been estimated using the initial rise and peak shape approaches by fitting the thermoluminescence (TL) glow peaks of the PPLG samples exposed to 20 Gy. The lifetime of the TL glow peak is also presented, which provides information on the stability of the TL signal at maximum temperatures. Raman, Photoluminescence (PL), and X-ray diffraction (XRD) spectra are being used to observe the structural changes that have occurred as a result of the radiation doses. These spectroscopies offer an understanding of the physical parameters that are related to the defects and taking part in the luminescence process. When all of the data are taken into account, it is anticipated that 0.3 mm PPLG is an effective material for dosimetry. The results of these lines of research are intended to educate the innovation of versatile graphite radiation dosimeters as a low-cost efficient system for radiation detection. The studied PPLG offers tissue equivalence as well as high spatial resolution, both are desirable criteria for a material to be used in the monitoring of ionising radiation or a variety of medical applications.

Studies of defect states and kinetic parameters of car windscreen for thermoluminescence retrospective dosimetry.

In case of any natural disasters or technical failures of nuclear facilities, the surrounding media including human beings may receive unexpected radiation exposures. In such a situation, there is no viable way to know how much radiation dose is received by human beings. Realizing that motorized vehicles are parked at fixed but increasing distances within the nuclear installation and industrial environment, this study investigates the kinetic parameters of readily available car windscreens which form the basis to be employed in post-accident dose reconstruction or for retrospective dosimetry. To understand the luminescence features of this crystalline media, a convenient thermoluminescence (TL) technique has been employed. Several well-defined theoretical models and methods were employed to calculate the kinetic parameters including the order of kinetics (b), activation energy (E) or trap depth, frequency factor (s) or escape probability and trap lifetime (τ), by analyzing the glow curves of the irradiated samples. The analysed trapping parameters indicate that the Toyota (E = 0.75-1.31 eV, s = 3.0E+6 - 3.7E+9 (s-1), τ = 6.9E+5 - 1.3E+14 s) and Honda (E = 0.95-1.68 eV, s = 2.1E+10 - 4.1E+13 (s-1), τ = 2.2E+9 - 3.1E+20 s) windscreen offer promising features for conventional TL dosimetry applications, while the obtained longer lifetime (τ = 6.8E+10 - 8.6E+29 s) or higher activation energy (E = 1.23-2.15 eV) for Proton brand windscreen indicates better stability or slow fading of the material, thus suitable for retrospective TL dosimetry. In addition, by assessing the area of deconvoluted micro-Raman spectra of windshield glasses in high-frequency regions, it has been observed the phenomenon of dose-dependent structural alterations and internal annealing of defects. This pattern is also consistent with those cyclical pattern observed in the intensity ratio of defect and graphite modes in the studies of carbon-rich media. Such common phenomena indicate the possibility of using the Raman microspectroscopy as a probe of radiation damage in silica-based media.

The potential of decorative building materials (marble) for retrospective thermoluminescence dosimetry.

Among the various types of decorative materials used in Bangladeshi dwellings, the marble/marble stone is one of the most common ones that used largely for enhancing the beauty and/or aristocracy of the dwelling environment. In this study, the most commonly used, six types of marble stones, have been analyzed for retrospective accident dosimetry. With the interest of characterizing several key thermoluminescence properties to examine their potentiality for dosimetry, annealing - irradiation - readout steps have been done chronologically which comprises the analysis of glow curves, relative sensitivity, dose dependence, repeatability and fading. Considering the various TL parameters, marble 'Carrara' imported from Italy present relatively better capability for reconstruction of radiation dose in the dose range of 10-50 Gy. From fading result, it is clear that for reconstruction of absorbed dose up to four weeks of post exposure, the marble 'Carrara' is found to be the most reliable media among the studied marble types. The Zeff values for the various marble samples are found to be in the range of 13.65-19.12, comparing favorably in replace of TLD-200 (Zeff = 16.3) which can be used for low-level environmental radiation dosimetry. Present work constitutes the first study to investigate the potentials of marble stone for reconstruction of absorbed dose in the range of 10-50 Gy dose.

Borosilicate glass 60Co high dose rate brachytherapy thermoluminescence dosimetry.

Brachytherapy is commonly used in treatment of cervical, prostate, breast and skin cancers, also for oral cancers, typically via the application of sealed radioactive sources that are inserted within or alongside the area to be treated. A particular aim of the various brachytherapy techniques is to accurately transfer to the targeted tumour the largest possible dose, at the same time minimizing dose to the surrounding normal tissue, including organs at risk. The dose fall-off with distance from the sources is steep, the dose gradient representing a prime factor in determining the dose distribution, also representing a challenge to the conduct of measurements around sources. Amorphous borosilicate glass (B2O3) in the form of microscope cover slips is recognized to offer a practicable system for such thermoluminescence dosimetry (TLD), providing for high-spatial resolution (down to < 1 mm), wide dynamic dose range, good reproducibility and reusability, minimal fading, resistance to water and low cost. Herein, investigation is made of the proposed dosimeter using a 1.25 MeV High Dose Rate (HDR) 60Co brachytherapy source, characterizing dose response, sensitivity, linearity index and fading. Analysis of the TL glow curves were obtained using the Tmax-Tstop method and first-order kinetics using GlowFit software, detailing the frequency factors and activation energy.

Indication of high lipid content in epithelial-mesenchymal transitions of breast tissues.

The epithelial-mesenchymal transition (EMT) is a crucial process in cancer progression and metastasis. Study of metabolic changes during the EMT process is important in seeking to understand the biochemical changes associated with cancer progression, not least in scoping for therapeutic strategies aimed at targeting EMT. Due to the potential for high sensitivity and specificity, Raman spectroscopy was used here to study the metabolic changes associated with EMT in human breast cancer tissue. For Raman spectroscopy measurements, tissue from 23 patients were collected, comprising non-lesional, EMT and non-EMT formalin-fixed and paraffin embedded breast cancer samples. Analysis was made in the fingerprint Raman spectra region (600-1800 cm-1) best associated with cancer progression biochemical changes in lipid, protein and nucleic acids. The ANOVA test followed by the Tukey's multiple comparisons test were conducted to see if there existed differences between non-lesional, EMT and non-EMT breast tissue for Raman spectroscopy measurements. Results revealed that significant differences were evident in terms of intensity between the non-lesional and EMT samples, as well as the EMT and non-EMT samples. Multivariate analysis involving independent component analysis, Principal component analysis and non-negative least square were used to analyse the Raman spectra data. The results show significant differences between EMT and non-EMT cancers in lipid, protein, and nucleic acids. This study demonstrated the capability of Raman spectroscopy supported by multivariate analysis in analysing metabolic changes in EMT breast cancer tissue.

Microscope cover-slip glass for TLD applications.

As a result of the various evolving needs, thermoluminescence dosimetry is constantly under development, with applications intended in environmental and personal radiation monitoring through to the sensing of radiotherapy and radiation processing doses. In radiotherapy dosimetry challenges include small-field profile evaluation, encompassing the fine beams of radiosurgery, evaluations confronting the steep dose gradients of electronic brachytherapy and the high dose rates of FLASH radiotherapy. Current work concerns the thermoluminescent dosimetric properties of commercial low-cost borosilicate glass in the form of thin (sub-mm to a few mm) plates, use being made of microscope cover-slips irradiated using clinical external-beam radiotherapy facilities as well as through use of 60Co gamma irradiators. In using megavoltage photons and MeV electrons, characterization of the dosimetric response has been made for cover-slips of thicknesses up to 4 mm. Reproducibility to within +/5% has been obtained. In particular, for doses up to 10 Gy, the borosilicate cover-slips have been demonstrated to have considerable potential for use in high spatial resolution radiotherapy dosimetry, down to 0.13 mm in present work, with a coefficient of determination in respect of linearity of >0.99 for the thinner cover-slips. Results are also presented for 0.13- and 1.00-mm thick cover slips irradiated to 60Co gamma-ray doses, initially in the range 5- to 25 Gy, subsequently extended to 5 kGy-25 kGy, again providing linear response.

Sub kGy photon irradiation alterations in graphite.

Present work concerns polymer pencil-lead graphite (PPLG) and the potential use of these in elucidating irradiation-driven structural alterations. The study provides detailed analysis of radiation-induced structural interaction changes and the associated luminescence that originates from the energy absorption. Thermally stimulated emission from the different occupied defect energy levels reflects the received radiation dose, different for the different diameter PPLGs. The PPLG samples have been exposed to photon irradiation, specifically x-ray doses ranging from 1 to 10 Gy, extended to 30-200 Gy through use of a60Co gamma-ray source. Trapping parameters such as order of kinetics, activation energy and frequency factor are estimated using Chen's peak-shape method for a fixed-dose of 30 Gy. X-ray diffractometry was used to characterize the crystal structure of the PPLG, the aim being to identify the degree of structural order, atomic spacing and lattice constants of the various irradiated PPLG samples. The mean atomic spacing and degree of structural order for the different diameter PPLG are found to be 0.3332 nm and 26.6° respectively. Photoluminescence spectra from PPLG arising from diode laser excitation at 532 nm consist of two adjacent peaks, 602 nm (absorption) and 1074 nm (emission), with mean energy band gap values within the range 1.113-1.133 eV.

Thermoluminescence response of X-ray irradiated commercial chalk.

Present research concerns the TL signal stored in chalk of the variety commercially available for writing on blackboards. Samples of this have been subjected to x-ray irradiation, the key dosimetric parameters investigated including dose and energy response, sensitivity, fading and glow curve analysis. Three types of chalk have been investigated, each in five different colours. The samples were annealed at 323 K prior to irradiation. For all three chalk types and all five colours, the dose response has been found linear over the investigated dose range, 0-9 Gy. Regardless of type or colour, photoelectric energy dependency is apparent at the low energy end down to the lowest investigated accelerating potential of 30 kV. Crayola (Yellow) has shown the greatest TL sensitivity, thus selection has been made to limit further analysis to this medium alone, specifically in respect of glow curve and fading study. In addition, elemental compositional and structural change characterizations were made for the same medium, utilizing Energy Dispersive X-Ray (EDX) and Raman spectroscopy, respectively.

Commercial kitchenware glass as a potential thermoluminescent media for retrospective dosimetry.

Study has been made of the thermoluminescence (TL) yield of various glass-based commercial kitchenware (Reko-China, Skoja-France, Godis-China, Glass Tum-Malaysia, Lodrat-France). Interest focuses on their potential for retrospective dosimetry. Use was made of a60Co gamma-ray irradiator, delivering doses in the range 2-10 Gy. Results for the various media show all the glassware brands to yield linearity of response against dose, with a lower limit of detection of ∼0.06 and ∼0.08 Gy for loose and compact powdered samples. Among all of the brands under study, the Lodrat glassware provides the greatest sensitivity, at 6.0 E+02 nC g-1 Gy-1 and 1.5E+03 nC g-1 Gy-1 for compact- and loose-powdered forms respectively. This is sufficiently sensitive to allow its use as a TL material for accident dosimetry (2 Gy being the threshold dose for the onset of a number of deterministic biological effects, including skin erythema and sterility). Energy Dispersive X-ray (EDX) analyses have been conducted, showing the presence of a number of impurities (including C, O, Na, Mg, Al, Si, Ca and Br). Fading of the irradiated glasses show the amount of better than 3% and 5% of the stored energy for both loose and compact powdered samples within 9 days post irradiation. As such, commercial kitchenware glass has the potential to act as relatively good TL material for gamma radiation dosimetry at accident levels. This is the first endeavour reporting the TL properties of low cost commercial kitchenware glasses for gamma-ray doses in the few Gy range, literature existing for doses from 8 Gy to 200 Gy.

Doped silica fibre thermoluminescence measurements of radiation dose in the use of 223Ra.

Using tailor-made sub-mm dimension doped-silica fibres, thermoluminescent dosimetric studies have been performed for α-emitting sources of 223RaCl2 (the basis of the Bayer Healthcare product Xofigo®). The use of 223RaCl2 in the palliative treatment of bone metastases resulting from late-stage castration-resistant prostate cancer focuses on its favourable uptake in metabolically active bone metastases. Such treatment benefits from the high linear energy transfer (LET) and associated short path length (<100µm) of the α-particles emitted by 223Ra and its decay progeny. In seeking to provide for in vitro dosimetry of the α-particles originating from the 223Ra decay series, investigation has been made of the TL yield of various forms of Ge-doped SiO2 fibres, including photonic crystal fibre (PCF) collapsed, PCF uncollapsed, flat and single-mode fibres. Irradiations of the fibres were performed at the UK National Physical Laboratory (NPL). Notable features are the considerable sensitivity of the dosimeters and an effective atomic number Zeff approaching that of bone, the glass fibres offering the added advantage of being able to be placed directly into liquid. The outcome of present research is expected to inform development of doped fibre dosimeters of versatile utility, including for applications as detailed herein.