Determination of equivalent breast phantoms for different age groups of Taiwanese women: an experimental approach.

PURPOSE: Polymethylmethacrylate (PMMA) slab is one of the mostly used phantoms for studying breast dosimetry in mammography. The purpose of this study was to evaluate the equivalence between exposure factors acquired from PMMA slabs and patient cases of different age groups of Taiwanese women in mammography. METHODS: This study included 3910 craniocaudal screen/film mammograms on Taiwanese women acquired on one mammographic unit. The tube loading, compressed breast thickness (CBT), compression force, tube voltage, and target/filter combination for each mammogram were collected for all patients. The glandularity and the equivalent thickness of PMMA were determined for each breast using the exposure factors of the breast in combination with experimental measurements from breast-tissue-equivalent attenuation slabs. Equivalent thicknesses of PMMA to the breasts of Taiwanese women were then estimated. RESULTS: The average +/- standard deviation CBT and breast glandularity in this study were 4.2 +/- 1.0 cm and 54% +/- 23%, respectively. The average equivalent PMMA thickness was 4.0 +/- 0.7 cm. PMMA slabs producing equivalent exposure factors as in the breasts of Taiwanese women were determined for the age groups 30-49 yr and 50-69 yr. For the 4-cm PMMA slab, the CBT and glandularity values of the equivalent breast were 4.1 cm and 65%, respectively, for the age group 30-49 yr and 4.4 cm and 44%, respectively, for the age group 50-69 yr. CONCLUSIONS: The average thickness of PMMA slabs producing the same exposure factors as observed in a large group of Taiwanese women is less than that reported for American women. The results from this study can provide useful information for determining a suitable thickness of PMMA for mammographic dose survey in Taiwan. The equivalence of PMMA slabs and the breasts of Taiwanese women is provided to allow average glandular dose assessment in clinical practice.

Development of an adjustable model breast for mammographic dosimetry assessment in Taiwanese women.

OBJECTIVE: The currently used model breast for mammographic dosimetry assessment lacks the flexibility to change dimensions. The aim of this study was to develop an adjustable model breast for mammographic dosimetry assessment of Taiwanese women. MATERIALS AND METHODS: A retrospective review of 4226 craniocaudal (CC) views was conducted. The geometry of the model breast was defined as a semielliptical cylinder. Breast parameters, including compressed breast thickness, chest wall-to-nipple distance, compressed breast width, and percentage glandular content, were measured and analyzed. To validate the adjustable model breast, 44 mammograms were obtained. The expected values from the adjustable model breast were compared with the measured values. RESULTS: The average values of compressed breast thickness, chest wall-to-nipple distance, compressed breast width, and percentage glandular content of the women studied were 4.1 cm, 6.9 cm, 16.9 cm, and 54%, respectively. Variations of chest wall-to-nipple distance, compressed breast width, and percentage glandular content can be expressed as functions of compressed breast thickness, and the adjustable model breast developed was based on compressed breast thickness. The average area of the CC view obtained is a factor of 0.81 lower than that defined by the American College of Radiology protocol. For validation, the difference in average values between the expected and measured did not exceed 0.5 cm in breast dimensions and 6% in percentage glandular content. CONCLUSION: Compressed breast thickness is useful for quantifying dimensions and percentage glandular content of a model breast. The adjustable model breast developed in this study can offer greater flexibility in the determination of breast dimensions for mammographic dosimetry assessment of Taiwanese women.

Biodistribution, toxicity and radiation dosimetry studies of the serotonin transporter radioligand 4-[18F]-ADAM in rats and monkeys.

PURPOSE: 4-[(18)F]-ADAM is a potent serotonin transport imaging agent. We studied its toxicity in rats and radiation dosimetry in monkeys before human studies are undertaken. METHODS: Single and multiple-dosage toxicity studies were conducted in Sprague-Dawley rats. Male and female rats were injected intravenously with 4-F-ADAM as a single dose of 1,023.7 microg/kg (1,000 times the human dose) or as five consecutive daily doses of 102.37 microg/kg (100 times the human dose). PET/CT scans were performed in seven Formosa Rock monkeys (four males and three females) using a Siemens Biograph scanner. After injection of 4-[(18)F]-ADAM (182+/-8 MBq), a low dose CT scan and a series of eight whole-body PET scans were performed. Whole-body images were acquired in 3-D mode. Time-activity data of source organs were used to calculate the residence times and estimate the absorbed radiation dose using OLINDA/EXM software. RESULTS: In the rats neither the single dose nor the five daily doses of 4-F-ADAM produced overt adverse effects clinically. In the monkeys the radiation doses received by most organs ranged between 7.1 and 35.7 microGy/MBq, and the urinary bladder was considered to be the critical organ. The effective doses extrapolated to male and female adult humans were 17.4 and 21.8 microSv/MBq, respectively. CONCLUSION: Toxicity studies in Sprague-Dawley rats and radiation dosimetry studies in Formosa Rock monkeys suggested that 4-[(18)F]-ADAM is safe for use in human PET imaging studies.

Prognostic factors in women with early stage small cell carcinoma of the uterine cervix.

Small cell carcinoma of the uterine cervix (SCCUC) is an uncommon, aggressive disease accounting for less than 5% of all cervical cancers. Due to its rarity, definitive treatment strategies have not been developed. Our aim was to analyze the clinical factors, treatment modalities, sites of relapse, and overall survival of women with early stage SCCUC and thus determine prognostic factors. The clinical records of 18 women diagnosed with stage IB1 to IIA SCCUC were reviewed, and patient characteristics and treatment modalities were analyzed to determine the prognostic factors for disease-free survival (DFS) and overall survival (OS). DFS and OS were 39% and 44% at 2 years. Lymph node metastasis was a significant prognostic factor of DFS. International Federation of Gynecology and Obstetrics (FIGO) stage and lymph node metastasis were significant prognostic factors of OS as determined by multivariate analysis (p < 0.05). Radical hysterectomy followed by adjuvant chemotherapy resulted in higher 2-year survival rates compared to radical hysterectomy followed by adjuvant radiotherapy (62.5% vs. 16.7%); however, the difference was not statistically significant due to the small sample size. FIGO stage and lymph node metastasis are significant indicators of OS in patients with early stage SCCUC. Further larger scale analysis is warranted to determine whether adjuvant chemotherapy may facilitate a better prognosis than adjuvant radiotherapy.

Effect of cigarette smoking on bone mineral density in healthy Taiwanese middle-aged men.

The effect of cigarette smoking in relation to bone mineral density (BMD) remains inconclusive, especially in middle-aged men. This cross-sectional study was conducted to examine the effect of smoking on BMD in 837 healthy Taiwanese males (532 never-smokers, 258 current smokers, 47 former smokers; aged 46-64 yr), recruited at their routine health examination. Subjects with suspected conditions affecting bone metabolism or receiving any medications affecting bone metabolism were excluded. BMD of the lumbar spine (LSBMD) and femoral neck (FNBMD) was measured with dual-energy X-ray absorptiometry. After adjustment for confounding variables (age, weight, physical activity, alcohol consumption, and caffeine intake), we found that the mean value of LSBMD was significantly (2.9%) lower in current smoker compared with never-smokers (p=0.024), but no significant difference was observed in FNBMD. No statistically significant association was observed between former smokers and never-smokers in any of the BMD sites, indicating that quitting smoking did have a positive effect on bone density. Compared with never-smokers, current heavy smokers who consumed at least 20 cigarettes/d (n=94) had 3.8% lower LSBMD (p=0.04), but no significant difference was observed in FNBMD. In the correlation analysis, the duration of smoking was negatively associated with LSBMD (r=-0.166, p=0.004), but no association was shown in FNBMD. Our results suggested that both smoking status and duration of smoking were deleterious factors on the bone density of the lumbar spine, and the effect was cumulative with duration and quantity.

Micro-column solid phase extraction to determine uranium and thorium in environmental samples.

Extraction chromatographic separation techniques based on U/TEVA and TEVA resins were utilized to separate uranium and thorium isotopes in complex matrices from environmental samples. This approach has the advantages of ease of quantitative analysis, small sample size, an absence of mixed waste solvents, complete separation of U/Th isotopes, acceptable chemical yields and good energy resolution in the alpha spectrum. The procedure for analyzing alpha-emitting isotopes of uranium and thorium in geothermal water from Peito, Taiwan, is illustrated in detail. It involves sample pre-concentration, filtration and separation by highly selective extraction chromatographic resins, followed by electroplating and alpha-spectroscopy. The analytical results show a chemical recovery exceeding 55% for U and 65% for Th, respectively, under optimized conditions. The efficient and cost-effective use of recyclable columns makes the analytical methods simple, accurate, rapid, reliable and robust.

On-line enhancement and separation of nanoparticles using capillary electrophoresis.

We describe a rapid, simple, and highly efficient capillary electrophoresis (CE)-based method for the analysis of nanoparticles (NPs). In this study, we used the reversed electrode polarity stacking mode (REPSM) of CE to assess the feasibility of enhancing the detection of Au NPs and Au/Ag NPs, optimizing parameters such as the length of time for which the REPSM was applied, the concentrations of the buffer and the sodium dodecylsulfate (SDS) surfactant, and the pH. Under the optimized on-line enhancement conditions [buffer: SDS (40 mM) and 3-cyclohexylamino-1-propanesulfonic acid (CAPS; 10 mM) at pH 10.0; applied voltage: 20 kV; REPSM applied for 24s], the detection limits of the Au NPs and Au/Ag NPs increased by ca. 30- and 140-fold, respectively. In addition, when the NPs were subjected to on-line enhancement and separation by CE using diode array detection (DAD), this approach allowed chemical characterization of the NP species. Our results suggest that such CE analyses will be useful for accelerating the rates of fabrication and characterization of future nanomaterials.

Analytical separation of Au/Ag core/shell nanoparticles by capillary electrophoresis.

This paper demonstrates that capillary electrophoresis (CE) can be employed for characterizing the sizes of a series of Au/Ag core/shell nanoparticles (NPs). We effected the CE separation of Au/Ag core/shell NPs using a mixed buffer of sodium dodecyl sulphate (SDS) (40 mM) and 3-(cyclohexylamino)propanesulfonic acid (10 mM) at pH 9.7 and an applied voltage of 20 kV. A linear relationship (R(2)>0.99) existed between the electrophoretic mobilities and the sizes of the Au/Ag core/shell NPs within the diameter range from 25 to 90 nm; the relative standard deviations of these electrophoretic mobilities were <0.9%. From the good correlation between the results obtained by CE and those provided by scanning electron microscopy, we confirmed that this CE method is a valid one for characterizing the sizes of Au/Ag core/shell NP samples. In addition, when the Au/Ag core/shell NPs were separated through CE and detected using an on-line photodiode array detector, this approach allowed the chemical characterization of the NP species. This CE approach should allow the rapid and cost-effective characterization of a number of future nanomaterials.

Studying the size/shape separation and optical properties of silver nanoparticles by capillary electrophoresis.

This paper describes the feasibility of employing capillary electrophoresis (CE) to separate silver particles in nanometer regimes. We have found that the addition of an anionic surfactant, sodium dodecyl sulphate (SDS), to the running electrolyte prevents coalescence of the silver particles during the process, which improves the separation performance; the concentration of SDS required for optimal silver nanoparticle separation is ca. 20 mM. By monitoring the electropherograms using a diode-array detection (DAD) system, we have also investigated the separation of suspended silver nanorods with respect to their shapes. Our results demonstrate that the combination of CE and DAD is a powerful one for the separation and characterization of various silver nanoparticles.

A positron emission tomography/computed tomography (PET/CT) acquisition protocol for CT radiation dose optimization.

BACKGROUND: In current combined positron emission tomography/computed tomography (PET/CT) systems, high-quality CT images not only increase diagnostic value by providing anatomic delineation of hyper- and hypometabolic tissues, but also shorten the acquisition time for attenuation correction compared with standard PET imaging. However, this technique potentially introduces more radiation burden to patients as a result of the higher radiation exposure from CT. METHODS: In this study, the radiation doses delivered from typical germanium-based and CT-based transmission scans were measured and compared using an anthropomorphic Rando Alderson phantom with insertions of thermoluminescent dosimeters. Image geometric distortion and quantified uptake values in PET images with different manipulating CT acquisition protocols for attenuation correction were also evaluated. RESULTS: It was found that radiation doses during germanium-based transmission scans were almost negligible, while doses from CT-based transmission scans were significantly higher. Using a lower radiation dose, the CT acquisition protocol did not significantly affect attenuation correction and anatomic delineation in PET. CONCLUSIONS: This study revealed the relation between image information and dose. The current PET/CT imaging acquisition protocol was improved by decreasing the radiation risks without sacrificing the diagnostic values.

Temporal resolving power of perfusion- and BOLD-based event-related functional MRI.

Functional magnetic resonance imaging (fMRI) based on both perfusion and blood oxygenation level-dependent (BOLD) contrasts has been widely applied in spatiotemporal mapping of the human brain function. Temporal resolving power of fMRI is limited by the smoothed hemodynamic response function dispersed from the neuronal activity. In this study, temporal modulation transfer functions were utilized to quantify the resolving powers of perfusion and BOLD fMR signals in time domain. The impulse response function was determined using brief visual stimulations and event-related image acquisition schemes. An important feature of arterial spin labeling techniques is that quantitative perfusion and BOLD signals could be simultaneously acquired. This simultaneous BOLD response may arise from signals that are more proximal to capillary beds, and its temporal resolution may be different from that of the typical BOLD response. Therefore, we assessed and compared the temporal resolving capabilities of perfusion, simultaneous BOLD, and the typical BOLD response obtained from the gradient echo EPI pulse sequence. Full-width-at-half-maximums of perfusion and simultaneous BOLD measurements were significantly smaller than that of BOLD ones (4.3+/- 0.6 s vs 5.5 +/- 0.9 s, p<0.02 and 4.7 +/- 1.3 s vs 5.5 +/- 0.9 s, p<0.01, respectively). The corresponding temporal resolving powers of perfusion and simultaneous BOLD signals were statistically better than that of BOLD signals (0.23 +/- 0.03 Hz vs 0.17 +/- 0.02 Hz, p<0.01 and 0.21 +/- 0.04 Hz vs 0.17 +/- 0.02 Hz, p<0.01, respectively). Our results showed that the typical BOLD response was significantly smoothed from the perfusion response, thus resulting in a degraded temporal resolving power. However, results from the simultaneous BOLD and perfusion measurements were not significantly different. Biophysical implications of the experimental outcomes were further investigated using a computer simulation based on the Balloon model. By fitting the measured data into the model, an apparently longer transit time was obtained for the typical BOLD signal (1.7 s), comparing to that for the simultaneous BOLD one (1.2 s). Therefore, the simultaneous BOLD signal was regarded as less susceptible to the variations from local draining veins. Combining the simulation result with the significantly discrepant resolving powers between the two BOLD signals, we speculated that the blurred effects from large vessels played a predominant role that further reduced the temporal resolution of the BOLD-based fMRI from the perfusion response.

Correlation between temporal response of fMRI and fast reaction time in a language task.

Correlation between behavioral parameters and fMRI responses can provide an advanced understanding of the neuronal processes. A lexical decision task was employed to examine the correlation between the reaction time (RT) and the temporal parameters in event-related BOLD responses. Word frequency was manipulated in the experiment. RTs for high-frequency, low-frequency and pseudowords were measured during fMRI (417 +/- 9 ms, 631 +/- 22 ms and 658 +/- 15 ms, respectively). For high-frequency words, RTs were significantly shorter than that for low-frequency and pseudowords (p < 0.0005). In the left inferior frontal region, the FWHM of the fMRI responses was significantly correlated with RT (p < 0.001), which may correspond to areas with sustained activation during the whole processing.

Study of induced thermoluminescence in CVD diamond film by low-energy X-rays.

For diamond film the one-hit model that is used to interpret low-energy X-ray thermoluminescence (TL) will require some modifications. After the films were irradiated with a superficial X-ray machine with different peak voltages, a two-compartment model with three parameters, the target size, the microscopic saturation factor and the high-LET saturation factor, was used to more precisely describe the TL response to X-ray with energies down to 10 kV. The microdosimetric distribution was calculated using single-event Monte Carlo code developed by authors together with EEDL cross-section data library. Some mechanistic insight into the physical aspect of radiation interaction with solid detectors can be obtained. The sensitive size in diamond was found to be about 15 nm. The saturation of one group of sublevels combined with the activation of another group of sublevels caused the relative efficiency to have a local minimum near 20 keV. The relative efficiency becomes higher below 10 keV, which is similar to the increasing relative biological effectiveness when the linear energy transfer passing through a biological system increases. The similarity made this material to be a molecular-scale dosimeter in the future.

Accuracy of the convolution/superposition dose calculation algorithm at the condition of electron disequilibrium.

Using Monte Carlo simulation and the convolution/superposition algorithm, this work examines percent depth dose curves of the central axis in an acrylic phantom (20 x 20 x 20 cm(3)) with variously sized air cavities (20 x 20 x 1.0, 20 x 20 x 2.0, 20 x 20 x 3.0, 20 x 20 x 4.0 and 20 x 20 x 4.95 cm(3) for study of longitudinal electron disequilibrium (ED) and 3.6 x3.6 x 4.95, 4.5 x 4.5 x 4.95, 5.4 x 5.4 x 4.95 and 20 x 20 x 4.95 cm(3) for study of lateral ED). Radiochromic film samples are also measured to verify the Monte Carlo results. The Monte Carlo simulation is performed using OMEGA/BEAM and DOSXYZ codes, and the convolution/superposition calculation relies on an ADAC commercial treatment planning system. Underestimating the dose kernel expansion leads to overestimating the dose of what was found in the air cavity of ED using the convolution/superposition algorithm. Consequently, the dose in the rebuild-up region is influenced. The influenced region is on the acrylic phantom surface to a depth of about 0.5 cm. The density scaling method of the convolution/superposition algorithm, applied to heterogeneous media, should be enhanced to account for the over-expansion of the dose kernel in the cavity of ED.

Indoor light on thermoluminescence of CVD diamond film used as a high-energy photon dosimeter.

The effect of light on polycrystalline diamond film that was produced by chemical vapor deposition and is used as a thermoluminescent dosimeter should be considered, although some researchers have indicated that such an effect was theoretically unlikely to happen. A 15 min exposure to a normal desk light bulb induces significant thermoluminescence (TL) comparable to a 0.5 Gy exposure to high-energy photons. This light-induced TL will be saturated within 2 h. The saturated TL intensity depends on the frequency of the light and the blue light dominates. The TL peak area at a temperature of 605 K is insensitive to light but is sensitive to high-energy photons. Another peak at about 410 K is caused by light only, because the TL from the ionization radiation at the same location is bleached. The effect of light could be easily distinguished by a numerical or an experimental method. Lamps with a green lampshade or pure red lights are suggested for use as indoor light sources. To reduce the effect of light, pre-heating treatment before readout is also suggested.

Surface dose with grids in electron beam radiation therapy.

This investigation attempts to solve the problem of the lack of skin-sparing effect in electron radiation therapy and to increase the tolerance of skin to radiation using the grid technique. Electron grid therapy involves the mounting of a Cerrobend grid in the electron cone. Film dosimetry was employed to measure the relative surface dose and the percentage depth dose profile of electron grid portals. Various grid hole diameters (d = 0.45, 1.0, 1.5 cm) and grid hole spacings (s = 0.4, 0.2 cm) were considered for electron beams from 6 to 14 MeV. Experimental results indicate that the electron grid technique can reduce the relative surface dose in electron radiation therapy. Degradations of the relative surface dose depend on the percentage of open area in the grid portal. A proper grid design allows the surface dose to be reduced and the range of nonhomogeneous doses to be limited to a depth at which the target volume can receive a homogeneous dose. The grid technique can lower the surface dose in electron radiation therapy.

Monte Carlo simulation of surface percent depth dose.

This work studied the surface percent depth dose of 6 and 15 MV X-rays, 10 x 10 cm2 and 20 x 20 cm2 fields by Monte Carlo simulation. The OMEGA/BEAM code, an EGS4 user code developed by the NRCC, was used. The linac, Siemens PRIMUS, was accurately modeled according to the ion chamber and CEA film measurement, and the phase space data generated from this linac were collected to simulate dose distribution in water. The water phantom had radius 30 cm and thickness 10 cm. The percent depth doses at zero depth, PDDsurface, for 6 MV X-rays were 13.85 +/- 0.11% and 23.21 +/- 0.20% for the 10 x 10 cm2 and 20 x 20 cm2 fields, respectively. For 15 MV X-rays, PDDsurface values were 8.83 +/- 0.07% and 18.60 +/- 0.12% for the 10 x 10 cm2 and 20 x 20 cm2 fields, respectively.

The effect of a metal hip prosthesis on the radiation dose in therapeutic photon beam irradiations.

Prostate and cervical cancer patients are often treated with external X-ray beams of bi-lateral incidence. Such treatment may incur some dose effect that cannot be predicted precisely in commercial treatment planning systems (TPS) for patients having undergone total hip replacement. This study performs a Monte Carlo (MC) simulation and an analytical calculation (convolution superposition algorithm which is implemented in ADAC TPS) of a 6 MV, 5 x 5 cm2 X-ray beam incident into water with the existence of hip prosthesis, e.g. Ti6A14V and CoCrMo alloy. The results indicate that ADAC TPS cannot precisely account for the scatter and backscatter radiation that a metal hip prosthesis causes. For percent depth dose (PDD) curves, the maximum underdosage of ADAC TPS up to 5mm above the interface between dense material and water is 5%, 20% and 27% for PDD(Bone), PDD(Ti) and PDD(Co), respectively. The dose re-buildup, which occurs behind the hip region, becomes more and more obvious for denser medium existed in water. Increasing inhomogeneity also enhances the underdosage of ADAC for greater depth (> 10cm), as the figures of nearly 2% in PDD(Bone), PDD(Ti) and 4-5% in PDD(Co) reveal. Overestimating the attenuated power of high-density non-water material in ADAC TPS causes this underdosage. For dose profiles, no significant differences were found in Profile(Bone) at any depth. Profile(Ti) reveals that MC slightly exceeds ADAC at off-axis position 1.0-2.0 cm. Profile(Co) reveals this more obviously. This finding means that scatter radiation from these denser materials is significant and cannot be predicted precisely in ADAC.

Quantitative ultrasound of the calcaneus in hemodialysis patients.

The aim of this study was to investigate the bone status of hemodialysis patients and identify factors that have influence on bone quality. Four hundred eighty-nine subjects (213 males and 276 females) on maintenance hemodialysis and 696 healthy subjects (309 men, 387 women) were enrolled in this study. Speed of sound (SOS), broadband ultrasound attenuation (BUA) and quantitative ultrasound index (QUI) were assessed by quantitative ultrasound (QUS) at the right calcaneus in both groups. Serum levels of intact parathyroid (iPTH), total alkaline phosphatase (ALP), calcium and phosphate were measured to determine their influence on bone status in hemodialysis patients. All QUS parameters were significantly lower in hemodialysis patients than in controls (p < 0.0001). Stepwise multiple linear regression analysis in male patients indicated that age, weight, calcium-phosphate product and ALP were significant predictors of QUS parameters (adjusted R(2) = 0.15 in SOS; adjusted R(2) = 0.17 in BUA and QUI). In female patients, same findings including number of parity were observed in SOS only (adjusted R(2) = 0.25 in SOS). In postmenopausal patients, the duration of menopause was significant negatively correlated with all QUS parameters (p < 0.01). In conclusion, patients on maintenance hemodialysis had additional risk of bone loss. Advanced age, low body weight, high calcium-phosphate product and high ALP level were important risk factors for deterioration of bone quality.

An improved synthesis of 4-[18F]-ADAM, a potent serotonin transporter imaging agent.

An improved synthesis of N,N-dimethyl-2-(2-amino-4-[(18)F]fluorophenylthio)benzylamine (4-[(18)F]-ADAM, 2) as a potent serotonin transporter (SERT) imaging agent is described. Molecular orbital (MO) calculation predicts that N,N-dimethyl-2-(2-nitro-4-trimethylammoniumtrifluoromethanesulfonylphenylthio)benzamide (8) is probably a better precursor than N,N-dimethyl-2-(2,4-dinitrophenylthio)benzylamine (1) for preparing 2. Radioligand 2 was synthesized by the reaction of either precursor 1 or precursor 8 with K[(18)F]/K(2.2.2) at 120 degrees C followed by reduction with BH(3) at 80 degrees C. The radiochemical yield (EOB) of 2 synthesized from precursor 1 and 8 was 5.7+/-2.4% (n=6) and 14.8+/-4.0% (n=5), respectively, in a synthesis time of 120 min from EOB. The specific activity of 2 was 3 Ci/micromol or 111 GBq/micromol (EOB). Thus, this new synthetic method has significantly improved the radiochemical yield of 4-[(18)F]-ADAM and makes this radioligand more accessible to PET Centers without a cyclotron.