Q-band electron paramagnetic resonance dosimetry in tooth enamel: biopsy procedure and determination of dose detection limit

High-frequency Q-band (37 GHz) electron paramagnetic resonance (EPR) dosimetry allows to perform fast (i.e., measurement time <15 min) dose measurements using samples obtained from tooth enamel mini-biopsy procedures. We developed and tested a new procedure for taking tooth enamel biopsy for such dose measurements. Recent experience with EPR dose measurements in Q-band using mini-probes of tooth enamel has demonstrated that a small amount of tooth enamel (2–10 mg) can be quickly obtained from victims of a radiation accident. Accurate dose assessments can further be carried out in a very short time to provide important information for medical treatment. Here, the Q-band EPR dose detection limit for 5 and 10 mg samples is estimated to be 367 and 248 mGy, respectively. These values are comparable to the critical parameters determined for conventional X-band EPR in tooth enamel.     

Quantifying the impact of µCT‐scanning of human fossil teeth on ESR age results

Fossil human teeth are nowadays systematically CT‐scanned by palaeoanthropologists prior to any further analysis. It has been recently demonstrated that this noninvasive technique has, in most cases, virtually no influence on ancient DNA preservation. However, it may have nevertheless an impact on other techniques, like Electron Spin Resonance (ESR) dating, by artificially ageing the apparent age of the sample. To evaluate this impact, we µCT‐scanned several modern enamel fragments following the standard analytical procedures employed by the Dental Anthropology Group at CENIEH, Spain, and then performed ESR dose reconstruction for each of them. The results of our experiment demonstrate that the systematic high‐resolution µCT‐scanning of fossil hominin remains introduces a nonnegligible X‐ray dose into the tooth enamel, equivalent to 15–30 Gy depending on the parameters used. This dose may be multiplied by a factor of ～8 if no metallic filter is used. However, this dose estimate cannot be universally extrapolated to any µCT‐scan experiment but has instead to be specifically assessed for each device and set of parameters employed. The impact on the ESR age results is directly dependent on the magnitude of the geological dose measured in fossil enamel but could potentially lead to an age overestimation up to 40% in case of Late Pleistocene samples, if not taken into consideration.     

Nitrogenase reduction of carbon disulfide: freeze-quench EPR and ENDOR evidence for three sequential intermediates with cluster-bound carbon moieties

Freeze-quenching of nitrogenase during reduction of carbon disulfide (CS(2)) was previously shown to result in the appearance of a novel EPR signal (g = 2.21, 1.99, and 1.97) not previously associated with any of the oxidation states of the nitrogenase metal clusters. In the present work, freeze-quench X- and Q-band EPR and Q-band (13)C electron nuclear double resonance (ENDOR) spectroscopic studies of nitrogenase during CS(2) reduction disclose the sequential formation of three distinct intermediates with a carbon-containing fragment of CS(2) bound to a metal cluster inferred to be the molybdenum-iron cofactor. Modeling of the Q-band (35 GHz) EPR spectrum of freeze-trapped samples of nitrogenase during turnover with CS(2) allowed assignment of three signals designated "a" (g = 2.035, 1.982, 1.973), "b" (g = 2.111, 2.002, and 1.956), and "c" (g = 2.211, 1. 996, and 1.978). Freezing samples at varying times after initiation of the reaction reveals that signals "a", "b", and "c" appear and disappear in sequential order. Signal "a" reaches a maximal intensity at 25 s; signal "b" achieves maximal intensity at 60 s; and signal "c" shows maximal intensity at 100 s. To characterize the intermediates, (13)CS(2) was used as a substrate, and freeze-trapped turnover samples were examined by Q-band (13)C ENDOR spectroscopy. Each EPR signal ("a", "b", and "c") gave rise to a distinct (13)C signal, with hyperfine coupling constants of 4.9 MHz for (13)C(a), 1. 8 MHz for (13)C(b), and 2.7 MHz for (13)C(c). Models for the sequential formation of intermediates during nitrogenase reduction of CS(2) are discussed.     

A method to differentiate between the levels of ESR signals induced by sunlight and by ionizing radiation in teeth from atomic bomb survivors

Electron spin resonance (ESR, or electron paramagnetic resonance, EPR) analysis of tooth enamel is an effective method for the retrospective estimation of individual radiation doses. One problem with this technique is that the observed ESR signal may include a contribution from ultraviolet (UV) light exposure from sunlight, especially in front teeth. Thus there has been a need to find ways to estimate the UV-light effect in the total signal so that the net ESR dose from ionizing radiation can be determined. To examine this issue, we measured 96 teeth of various types, but with buccal and lingual parts measured separately, from a control group of atomic bomb survivors (estimated dose <5 mGy). We found that, except for molars, the mean ESR-estimated dose for the buccal halves was, on average, nearly twice that from the lingual side, which indicates that the UV-light-induced lingual dose equals the difference between the two halves. Using these corrections for UV-light exposure to front teeth that had been exposed to both ionizing radiation and UV light, it was found that the estimated radiation doses closely approximated the previously estimated ESR dose to molars from the same donors or the estimated dose arrived at with cytogenetic methods. We concluded that, when using ESR to estimate radiation dose, measuring molars is the first choice, but if only front teeth are available, separate measurements to the buccal and lingual parts can provide an estimation of the mean UV-light contribution to the ESR-determined dose.     

Estimation of background radiation doses for the Peninsular Malaysia’s population by ESR dosimetry of tooth enamel

Background radiation dose is used in dosimetry for estimating occupational doses of radiation workers or determining radiation dose of an individual following accidental exposure. In the present study, the absorbed dose and the background radiation level are determined using the electron spin resonance (ESR) method on tooth samples. The effect of using different tooth surfaces and teeth exposed with single medical X-rays on the absorbed dose are also evaluated. A total of 48 molars of position 6–8 were collected from 13 district hospitals in Peninsular Malaysia. Thirty-six teeth had not been exposed to any excessive radiation, and 12 teeth had been directly exposed to a single X-ray dose during medical treatment prior to extraction. There was no significant effect of tooth surfaces and exposure with single X-rays on the measured absorbed dose of an individual. The mean measured absorbed dose of the population is 34 ± 6.2 mGy, with an average tooth enamel age of 39 years. From the slope of a regression line, the estimated annual background dose for Peninsular Malaysia is 0.6 ± 0.3 mGy y⁻¹. This value is slightly lower than the yearly background dose for Malaysia, and the radiation background dose is established by ESR tooth measurements on samples from India and Russia.     

ESR characteristics of one-electron reduced thymine in monomer, oligomer, and polymer derivatives

ESR spectra have been recorded of one-electron reduced thymine (T.-) trapped in a variety of thymine derivatives and matrices. Most of the data were taken at Q-band microwave frequencies on 12 M LiCl glasses containing oligo(T) varying from 2 to 18 nucleotides in length. A nonlinear least-squares simulation program was used to simulate the ESR spectra, resulting in a parameter set that can be used to generate T.- powder spectra at X- and Q-band frequencies. Although it is not possible to identify a unique set of g and hyperfine tensors, it is possible to establish boundaries for these interactions and propose a parameter set that is likely and reasonable. As the trapping molecule is varied from TdR to oligo(T) to poly(T), there are small changes in the interaction parameters. The changes are not large enough to account for the differences between hfc values determined from DNA fiber data versus hyperfine coupling values measured from monomer systems. The analysis also indicates that, in these systems, T.- does not protonate between 4 and 77 K at neutral pH.     

Molecular motion and order in oriented lipid multibilayer membranes evaluated by simulations of spin label ESR spectra. Effects of temperature, cholesterol and magnetic field.

A simulation method to interpret electron spin resonance (ESR) of spin labelled amphiphilic molecules in oriented phosphatidylcholine multibilayers in terms of a restricted motional model is presented. Order and motion of the cholestane spin label (3-spiro-doxyl-5alpha-cholestane) incorporated into egg yolk phosphatidylcholine, dipalmitoylphosphatidylcholine and dimyristoylphosphatidylcholine, pure and in mixture with cholesterol, were studied at various temperatures. With egg yolk phosphatidylcholine identical sets of motional parameters were obtained from simulations of ESR spectra obtained at three microwave frequencies (X-, K- and Q-band). With dipalmitoylphosphatidylcholine and dimyristoylphosphatidylcholine analyses of the spectra show that phase transitions occur in samples containing up to 30 mol % cholesterol. The activation energy for the motion of the spin label is about three times larger above than below the phase transition, indicating a more collective motion in the lipid crystalline state than in the gel state. In the liquid crystalline state the activation energy is larger in the pure phosphatidylcholines than with cholesterol added. Additions of cholesterol to egg phosphatidylcholine induces a higher molecular order but does not appreciably affect correlation times. This is in contrast to dipalmitoylphosphatidylcholine where both order and correlation times are affected by the presence of cholesterol. The activation energies follow the same order as the transition temperatures: dipalmitoylphosphatidylcholine greater than dimyristoylphosphatidylcholine greater than egg yokd phosphatidylcholine, suggesting a similar order of the cooperativity of the motion of the lipid molecules. Magnetic field-induced effects on egg phosphatidylcholine multibilayers were found at Q-band measurements above 40 degrees C. The cholestane spin label mimics order and motion of cholesterol molecule incorporated into the lipid bilayers. This reflects order and motion of the portions of the lipid molecules on the same depth of the bilayer as the rigid steroid portions of the intercalated molecules.     

Comparative X- and Q-band EPR study of radiation-induced radicals in tooth enamel

Human tooth enamel blocks and powders that were either unheated or heated prior to X irradiation at room temperature were investigated by means of Q-band electron paramagnetic resonance (EPR). It was found that the EPR spectra of unheated human tooth enamel consist mainly of two different anisotropic signals, as was suggested previously from an X-band study of analogous samples. In the present study, the two radical contributions could be differentiated convincingly by comparing the anisotropic Q-band spectra of heated and unheated enamel blocks. One type of is probably located in the bulk of the apatitic microcrystallites that constitute the enamel, and it appears in both heated and unheated samples. The other type is presumably located in an intercrystallite position and appears mainly in the unheated samples. Clear differences between g values in the Q-band spectra of heated and unheated enamel suggest that the radicals in the bulk exhibit larger g anisotropy than those in intercrystallite positions. Isotropic signals and contributions that may be from and radicals have also been detected. However, the present work focuses mainly on the signals and discusses potential and/or real difficulties that may be encountered in applications of EPR dosimetry using calcified tissues.     

Measurement of absorbed doses from X-ray baggage examinations to tooth enamel by means of ESR and glass dosimetry

The contribution of radiation from X-ray baggage scans at airports on dose formation in tooth samples was investigated by electron spin resonance (ESR) dosimetry and by glass dosimetry. This was considered important, because tooth samples from population around the Semipalatinsk Nuclear Test Site (SNTS), Kazakhstan, had been transported in the past to Hiroshima University for retrospective dose assessment of these residents. Enamel samples and glass dosimeters were therefore examined at check-in time at Kansai airport (Osaka, Japan), Dubai airport (Dubai, United Arab Emirates) and Domodedovo airport (Moscow, Russia). These airports are on the route from Kazakhstan to Japan. Three different potential locations of the samples were investigated: in pocket (without X-ray scans), in a small bag (with four X-ray scans) and in large luggage (with two X-ray scans). The doses obtained by glass and ESR dosimetry methods were cross-compared. As expected, doses from X-ray examinations measured by glass dosimetry were in the μGy range, well below the ESR detection limit and also below the doses measured in enamel samples from residents of the SNTS.     

Camel molar tooth enamel response to gamma rays using EPR spectroscopy

Tooth enamel samples from molar teeth of camel were prepared using a combined procedure of mechanical and chemical tooth treatment. Based on electron paramagnetic resonance (EPR) spectroscopy, the dose response of tooth enamel samples was examined and compared to that of human enamel. The EPR dose response of the tooth enamel samples was obtained through irradiation to gamma doses from 1 Gy up to 100 kGy. It was found that the radiation-induced EPR signal increased linearly with gamma dose for all studied tooth enamel samples, up to about 15 kGy. At higher doses, the dose response curve leveled off. The results revealed that the location of the native signal of camel tooth enamel was similar to that of enamel from human molars at 2.00644, but different from that of enamel from cows and goats. In addition, the peak-to-peak width (ΔH _pp) for human and camel molar teeth was similar. It was also found that the response of camel enamel to gamma radiation was 36% lower than that of human enamel. In conclusion, the results indicate the suitability of camel teeth for retrospective gamma dosimetry.     

北京猿人第一个头盖骨及其遗址堆积层年代的电子自旋共振测年研究

裴文中教授在60年前发现的北京猿人第一个头盖骨埋藏的确切年代至今没有获得。本文通过对北京猿人共生的古脊椎动物牙化石进行电子自旋共振(ESR)测年,分别获得了第11层北京猿人第一个头盖骨埋藏的年代为578千年,第8—9层和第3—4层北京猿人埋藏的年代分别为418千年和282千年。依据ESR、U系,TL,FT和古地磁等测年结果,本文推荐了北京猿人洞的洞穴堆积层(周口店组Q_2)年代表供读者参考。     

Characterization of phenolic pellets for ESR dosimetry in photon beam radiotherapy

This work deals with the dosimetric features of a particular phenolic compound (IRGANOX 1076^®) for dosimetry of clinical photon beams by using electron spin resonance (ESR) spectroscopy. After the optimization of the ESR readout parameters (namely modulation amplitude and microwave power) to maximise the signal without excessive spectrum distortions, basic dosimetric properties of laboratory-made phenolic dosimeters in pellet form, such as reproducibility, dose–response, sensitivity, linearity and dose rate dependence were investigated. The dosimeters were tested by measuring the depth dose profile of a 6 MV photon beam. A satisfactory intra-batch reproducibility of the ESR signal of the manufactured dosimeters was obtained. The ESR signal proved to increase linearly with increasing dose in the investigated dose range 1–13 Gy. The presence of an intrinsic background signal limits the minimum detectable dose to a value of approximately 0.6 Gy. Reliable and accurate assessment of the dose was achieved, independently of the dose rate. Such characteristics, together with the fact that IRGANOX 1076^® is almost tissue-equivalent, and the stability of the ESR signal, make these dosimeters promising materials for ESR dosimetric applications in radiotherapy.     

Electron paramagnetic resonance radiation dose assessment in fingernails of the victim exposed to high dose as result of an accident

In this paper, we report results of radiation dose measurements in fingernails of a worker who sustained a radiation injury to his right thumb while using 130 kVp X-ray for nondestructive testing. Clinically estimated absorbed dose was about 20–25 Gy. Electron paramagnetic resonance (EPR) dose assessment was independently carried out by two laboratories, the Naval Dosimetry Center (NDC) and French Institut de Radioprotection et de Sûreté Nucléaire (IRSN). The laboratories used different equipments and protocols to estimate doses in the same fingernail samples. NDC used an X-band transportable EPR spectrometer, e-scan produced by Bruker BioSpin, and a universal dose calibration curve. In contrast, IRSN used a more sensitive Q-band stationary spectrometer (EMXplus) with a new approach for the dose assessment (dose saturation method), derived by additional dose irradiation to known doses. The protocol used by NDC is significantly faster than that used by IRSN, nondestructive, and could be done in field conditions, but it is probably less accurate and requires more sample for the measurements. The IRSN protocol, on the other hand, potentially is more accurate and requires very small amount of sample but requires more time and labor. In both EPR laboratories, the intense radiation-induced signal was measured in the accidentally irradiated fingernails and the resulting dose assessments were different. The dose on the fingernails from the right thumb was estimated as 14 ± 3 Gy at NDC and as 19 ± 6 Gy at IRSN. Both EPR dose assessments are given in terms of tissue kerma. This paper discusses the experience gained by using EPR for dose assessment in fingernails with a stationary spectrometer versus a portable one, the reasons for the observed discrepancies in dose, and potential advantages and disadvantages of each approach for EPR measurements in fingernails.     

Long-term ozone exposure of potato: free radical content and leaf injury analysed by Q-band ESR spectroscopy and image analysis

This paper presents Q-band electron spin resonance (ESR) studies on free radicals (FR) generated in potato leaves exposed to different O₃ levels in open-top chambers (OTC), together with a quantitative study of the relationship between FR signal intensity and area of potato leaf damage. The advantages of Q-band when compared to X-band ESR spectroscopy are analysed, the main advantage being an absence of overlapping between Mn(II) and FR signals, allowing a quantitative study of FR signal intensity. This study also reports on a graphical method developed to quantitatively measure the damaged area on leaves caused by ozone exposure. Results indicate a direct relationship between FR signal intensity (measured as area under the signal) and percentage of O₃ damage and clearly demonstrate a close relationship between visible ozone-induced symptoms and permanent FR concentration in potato leaves.     

Direct ESR dating of a Pliocene hominin from Swartkrans

Two fragments of a hominin tooth (Australopithecus robustus) and two bovid teeth from the Hanging Remnant of the Swartkrans Formation were analysed with ESR. Research was complicated by the fact that the samples came from a curated collection and their precise provenance is unknown. The environmental dose rate was reconstructed by a series of in situ gamma spectrometric measurements and elemental analyses of a range of sediment samples. U-series isotopic analyses indicated that each of the teeth had a significantly different uranium uptake history, rendering the assumptive early U-uptake and linear U-uptake models ineffective. ESR and U-series data were combined to calculate open system ages, resulting in a best estimate of 1630+/-160 ka for the Hanging Remnant. An open-system model which provides the maximum age for given U-series and ESR measurements yielded an estimate of about 2100 ka.Two bovid teeth from Member 2, previously estimated to be between 1.0 and 2.0 Ma, yielded age estimates of between about 100 and 200 ka. No known geochemical processes are likely to explain this severe age underestimation. We conclude that these samples are of Middle to Upper Pleistocene age and their presence in Member 2 was either due to reworking or inadequate stratigraphical discrimination of these deposits.     

Studies on human uterine cervix and rat uterus using S-, X- and Q-band electron-spin-resonance spectroscopy.

In previous studies we have reported on the detection of a strong e.s.r. signal in samples of normal human cervix; the signal is much reduced or absent in samples of invasive cancer of the cervix. In order to identify the species responsible for the strong signal, we have used X-, S- and Q-band e.s.r. spectroscopy. The major signal that is detectable in ground-up samples of cervix preserved at -196 degrees C has features consistent with the presence of a peroxy free radical. Good agreement with the experimental findings was obtained by computer simulation, using values for the g-tensor of gx = 2.002, gy = 2.005 and gz = 2.036. The peroxy radical is produced on grinding the normal cervix samples to a powder under liquid N2, and appears to be formed by modification of a pre-existing oxygen-containing complex. Control experiments eliminated the possibility that the strong signals seen in frozen powders prepared from normal cervix were artefacts only of the grinding procedure. Experiments with rats in vivo and with cervix samples in vitro are consistent with the conclusion that the peroxy radical is formed by disturbing the cyclo-oxygenase system that is involved in prostaglandin synthesis.     

The Pech-de-l'Azé I Neandertal child: ESR, uranium-series, and AMS 14C dating of its MTA type B context

The Pech-de-l'Azé I skull and mandible are included in the juvenile Neandertal remains from Europe. However, some preserved features in the cranial skeleton seem to distinguish the specimen from other Neandertal children. Unfortunately, the stratigraphic position and dating of this child has never been clear. Our recent work on unpublished archives show that the Pech-de-l'Azé I Neandertal child was discovered at the bottom of layer 6, attributed to the Mousterian of Acheulean tradition type B. These skull and mandible are the first diagnostic human remains (aside from an isolated tooth) attributed to the Mousterian of Acheulian tradition (MTA) type B. Consequently, we confirm that Neandertals were the makers of this Mousterian industry, which is characterized by unusual high frequencies of Upper Paleolithic type tools, elongated blanks and blades. We were able to date the context of the hominid remains by dating layer 6 and the layers above and beneath it using ESR, coupled ESR/(230)Th/(234)U (coupled ESR/U-series), and AMS (14)C. Coupled ESR/U-series results on 16 mammalian teeth constrain the age of the uppermost layer 7 to 41-58ka, and layer 6 to 37-51ka. The wide spread in each age estimate results mainly from uncertainties in the gamma-dose rate. These ages are concordant with AMS (14)C ages of two bones coming from the top of layer 6, which provide dates of about 41.7-43.6ka cal BP. A combination of stratigraphic arguments and dating results for layers 6 and 7 show that the Neandertal child cannot be older than 51ka or younger than 41ka. The lowermost layer 4 is shown to be older than 43ka by the principle of superposition and ESR dating in the immediately overlying layer 5. This study shows that the MTA type B had been manufactured by Neandertals before the arrival of anatomically modern humans in the local region. Additionally, by providing a firm chronological framework for the specific morphometric the features of Pech-de-l'Azé I Neandertal child, this study is a new step toward the understanding of temporal and spatial changes in the ontogenesis of Neandertals in south-western Europe during oxygen isotope stages 5-3.     

Study of dental prostheses influence in radiation therapy

Dental prostheses made of high density material contribute to modify dose distribution in head and neck cancer treatment. Our objective is to quantify dose perturbation due to high density inhomogeneity with experimental measurements and Monte Carlo simulations.Firstly, measurements were carried in a phantom representing a human jaw with thermoluminescent detectors (GR200A) and EBT2 Gafchromic films in the vicinity of three samples: a healthy tooth, a tooth with amalgam and a Ni–Cr crown, irradiated in clinical configuration. Secondly, Monte Carlo simulations (BEAMnrc code) were assessed in an identical configuration.Experimental measurements and simulation results confirm the two well-known phenomena: firstly the passage from a low density medium to a high density medium induces backscattered electrons causing a dose increase at the interface, and secondly, the passage from a high density medium to a low density medium creates a dose decrease near the interface. So, the results show a 1.4% and 23.8% backscatter dose rise and attenuation after sample of 26.7% and 10.9% respectively for tooth with amalgam and crown compared to the healthy tooth.Although a tooth with amalgam has a density of about 12–13, the changes generated are not significant. However, the results for crown (density of 8) are very significant and the discordance observed may be due to calculation point size difference 0.8 mm and 0.25 mm respectively for TLD and Monte Carlo. The use of Monte Carlo simulations and experimental measurements provides objective evidence to evaluate treatment planning system results with metal dental prostheses.     

Molecular motion and order in oriented lipid multibilayer membranes evaluated by simulations of spin label ESR spectra. Effects of temperature,cholesterol and magnetic field

A simulation method to interpret electron spin resonance (ESR) of spin labelled amphiphilic molecules in oriented phosphatidylcholine multibilayers in terms of a restricted motional model is presented. Order and motion of the cholestane spin label (3-spiro-doxyl-5α-cholestane) incorporated into egg yolk phosphatidylcholine, dipalmitoylphosphatidylcholine and dimyristoylphosphatidylcholine, pure and in mixture with cholesterol, were studied at various termperatures. With egg yolk phosphatidylcholine identical sets of motional parameters were obtained from simulations of ESR spectra obtained at three microwave frequencies (X-, K- and Q-band). With dipalmitoylphosphatidylcholine and dimyristoylphosphatidylcholine analyses of the spectra show that phase transitions occur in samples containing up to 30 mol % cholesterol. The activation energy for the motion of the spin label is about three times larger above than below the phase transition, indicating a more collective motion in the liquid crystalline state than in the gel state. In the liquid crystalline state the activation energy is larger in the pure phosphatidylcholines than with cholesterol added. Additions of cholesterol to egg phosphatidylcholine induces a higher molecular order but does not appreciably affect correlation times. This is in contrast to dipalmitoylphosphatidylcholine where both order and correlation times are affected by the presence of cholesterol. The activation energies follow the same order as the transition temperatures: dipalmitoylphosphatidylcholine > dimyristoylphosphatidylcholine > egg yolk phosphatidylcholine, suggesting a similar order of the cooperativity of the motion of the lipid molecules. Magnetic field-induced effects on egg phosphatidylcholine multibilayers.