Hyperfractionation compared to standard fractionation in intensity-modulated radiation therapy for patients with locally advanced recurrent nasopharyngeal carcinoma.

We prospectively studied the efficacy and safety of hyperfractionated intensity-modulated radiation therapy (HF-IMRT) and compared to a historical cohort treated with standard fractionation (SF-IMRT) in patients with locally advanced recurrent (rT3-T4, rN0-N1, M0) nasopharyngeal carcinoma (NPC). Ten patients were treated with induction chemotherapy followed by HF-IMRT (64.8 Gy/54 fr/5.5 weeks) concurrent with weekly cisplatin. They were compared to another ten patients treated with induction chemotherapy followed by SF-IMRT (60 Gy/30 fr/6 weeks) concurrent with weekly cisplatin. After a median follow-up duration of 44.6 months, we demonstrated that the median local failure-free survival (LFFS) showed a trend in favor of HF-IMRT (28.2 vs. 16.6 months, p = 0.164). Overall survival (OS) (34.8 vs. 35.5 months, p = 0.603) was not different between the two groups. Treatment-related hemorrhage was slightly less with HF-IMRT (30.0 vs. 0 %), reaching marginal significance (p = 0.060). Judging from our study results, HF-IMRT offered a marginally better LFFS and an apparently more favorable toxicity profile compared to SF-IMRT in locally advanced recurrent NPC.

Quantifying variability of intrafractional target motion in stereotactic body radiotherapy for lung cancers.

In lung stereotactic body radiotherapy (SBRT), variability of intrafractional target motion can negate the potential benefits of four-dimensional (4D) treatment planning that aims to account for the dosimetric impacts of organ motion. This study used tumor motion data obtained from CyberKnife SBRT treatments to quantify the reproducibility of probability motion function (pmf) of 37 lung tumors. The reproducibility of pmf was analyzed with and without subtracting the intrafractional baseline drift from the original motion data. Statistics of intrafractional tumor motion including baseline drift, target motion amplitude and period, were also calculated. The target motion amplitude significantly correlates with variations (1SD) of motion amplitude and baseline drift. Significant correlation between treatment time and variations (1 SD) of motion amplitude was observed in anterior-posterior (AP) motion, but not in craniocaudal (CC) and left-right (LR) motion. The magnitude of AP and LR baseline drifts significantly depend on the treatment time, while the CC baseline drift does not. The reproducibility of pmf as a function of time can be well described by a two-exponential function with a fast and slow component. The reproducibility of pmf is over 60% for the CC motion and over 50% for the AP and LR motions when baseline variations were subtracted from the original motion data. It decreases to just over 30% for the CC motion and about 20% for the AP and LR motion, otherwise. 4D planning has obvious limitations due to variability of intrafractional target motion. To account for potential risks of overdosing critical organs, it is important to simulate the dosimetric impacts of intra- and interfractional baseline drift using population statistics obtained from SBRT treatments.

Evaluation of dose prediction error and optimization convergence error in four-dimensional inverse planning of robotic stereotactic lung radiotherapy.

Inverse optimization of robotic stereotactic lung radiotherapy is typically performed using relatively simple dose calculation algorithm on a single instance of breathing geometry. Variations of patient geometry and tissue density during respiration could reduce the dose accuracy of these 3D optimized plans. To quantify the potential benefits of direct four-dimensional (4D) optimization in robotic lung radiosurgery, 4D optimizations using 1) ray-tracing algorithm with equivalent path-length heterogeneity correction (4EPL(opt)), and 2) Monte Carlo (MC) algorithm (4MC(opt)), were performed in 25 patients. The 4EPL(opt) plans were recalculated using MC algorithm (4MC(recal)) to quantify the dose prediction errors (DPEs). Optimization convergence errors (OCEs) were evaluated by comparing the 4MC(recal) and 4MC(opt) dose results. The results were analyzed by dose-volume histogram indices for selected organs. Statistical equivalence tests were performed to determine the clinical significance of the DPEs and OCEs, compared with a 3% tolerance. Statistical equivalence tests indicated that the DPE and the OCE are significant predominately in GTV D98%. The DPEs in V20 of lung, and D2% of cord, trachea, and esophagus are within 1.2%, while the OCEs are within 10.4% in lung V20 and within 3.5% in trachea D2%. The marked DPE and OCE suggest that 4D MC optimization is important to improve the dosimetric accuracy in robotic-based stereotactic body radiotherapy, despite the longer computation time.

Patient dosimetry for 90Y selective internal radiation treatment based on 90Y PET imaging.

Until recently, the radiation dose to patients undergoing the 90Y selective internal radiation treatment (SIRT) procedure is determined by applying the partition model to 99mTc MAA pretreatment scan. There can be great uncertainty in radiation dose calculated from this approach and we presented a method to compute the 3D dose distributions resulting from 90Y SIRT based on 90Y positron emission tomography (PET) imaging. Five 90Y SIRT treatments were retrospectively analyzed. After 90Y SIRT, patients had 90Y PET/CT imaging within 6 hours of the procedure. To obtain the 3D dose distribution of the patients, their respective 90Y PET images were convolved with a Monte Carlo generated voxel dose kernel. The sensitivity of the PET/CT scanner for 90Y was determined through phantom studies. The 3D dose distributions were then presented in DICOM RT dose format. By applying the linear quadratic model to the dose data, we derived the biologically effective dose and dose equivalent to 2 Gy/fraction delivery, taking into account the spatial and temporal dose rate variations specific for SIRT. Based on this data, we intend to infer tumor control probability and risk of radiation induced liver injury from SIRT by comparison with established dose limits. For the five cases, the mean dose to target ranged from 51.7 ± 28.6 Gy to 163 ± 53.7 Gy. Due to the inhomogeneous nature of the dose distribution, the GTVs were not covered adequately, leading to very low values of tumor control probability. The mean dose to the normal liver ranged from 21.4 ± 30.7 to 36.7 ± 25.9 Gy. According to QUANTEC recommendation, a patient with primary liver cancer and a patient with metastatic liver cancer has more than 5% risk of radiotherapy-induced liver disease (RILD).

Dosimetric evaluation of four-dimensional dose distributions of CyberKnife and volumetric-modulated arc radiotherapy in stereotactic body lung radiotherapy.

Advanced image-guided stereotatic body lung radiotherapy techniques using volumetric-modulated arc radiotherapy (VMAT) with four-dimensional cone-beam computed tomography (4D CBCT) and CyberKnife with real-time target tracking have been clinically implemented by different authors. However, dosimetric comparisons between these techniques are lacking. In this study, 4D CT scans of 14 patients were used to create VMAT and CyberKnife treatment plans using 4D dose calculations. The GTV and the organs at risk (OARs) were defined on the end-exhale images for CyberKnife planning and were then deformed to the midventilation images (MidV) for VMAT optimization. Direct 4D Monte Carlo dose optimizations were performed for CyberKnife (4D(CK)). Four-dimensional dose calculations were also applied to VMAT plans to generate the 4D dose distributions (4D(VMAT)) on the exhale images for direct comparisons with the 4D(CK) plans. 4D(CK) and 4D(VMAT) showed comparable target conformity (1.31 ± 0.13 vs. 1.39 ± 0.24, p = 0.05). GTV mean doses were significantly higher with 4D(CK). Statistical differences of dose volume metrics were not observed in the majority of OARs studied, except for esophagus, with 4D(VMAT) yielding marginally higher D1% than 4D(CK). The normal tissue volumes receiving 80%, 50%, and 30% of the prescription dose (V80%, V50%, and V30%) were higher with 4D(VMAT), whereas 4D(CK) yielded slightly higher V10% in posterior lesions than 4D(VMAT). VMAT resulted in much less monitor units and therefore greater delivery efficiency than CyberKnife. In general, it was possible to produce dosimetrically acceptable plans with both techniques. The selection of treatment modality should consider the dosimetric results as well as the patient's tolerance of the treatment process specific to the SBRT technique.

Investigation of four-dimensional (4D) Monte Carlo dose calculation in real-time tumor tracking stereotatic body radiotherapy for lung cancers.

PURPOSE: To investigate the dosimetric variations and radiobiological impacts as a consequence of delivering treatment plans of 3D nature in 4D manner based on the 4D Monte Carlo treatment planning framework implemented on Cyberknife. METHODS: Dose distributions were optimized on reference 3D images at end of exhale phase of a 4DCT dataset for 25 lung cancer patients treated with 60 Gy∕3Fx or 48 Gy∕4Fx. Deformable image registrations between individual 3DCT images to the reference 3DCT image in the 4DCT study were performed to interpolate doses calculated on multiple anatomical geometries back on to the reference geometry to compose a 4D dose distribution that included the tracking beam motion and organ deformation. The 3D and 4D dose distributions that were initially calculated with the equivalent path-length (EPL) algorithm (3D(EPL) dose and 4D(EPL) dose) were recalculated with the Monte Carlo algorithm (3D(MC) dose and 4D(MC) dose). Dosimetric variations of V(60Gy∕48Gy) and D(99) of GTV, mean doses to the lung and the heart and maximum dose (D(1)) of the spinal cord as a consequence of tracking beam motion in deforming anatomy, dose calculation algorithm, and both were quantified by the relative change from 4D(MC) to 3D(MC) doses, from 4D(MC) to 4D(EPL) doses, and from 4D(MC) to 3D(EPL) doses, respectively. RESULTS: Comparing 4D(MC) to 3D(EPL) plans, V(60Gy ∕ 48Gy) and D(99) of GTV decreased considerably by 13 ± 22% (mean ± 1SD) and 9.2 ± 5.5 Gy but changes of normal tissue doses were not more than 0.5 Gy on average. The generalized equivalent uniform dose (gEUD) and tumor control probability (TCP) were reduced by 14.3 ± 8.8 Gy and 7.5 ± 5.2%, and normal tissue complication probability (NTCP) for myelopathy and pericarditis were close to zero and NTCP for radiation pneumonitis was reduced by 2.5% ± 4.1%. Comparing 4D(MC) to 4D(EPL) plans found decreased V(60Gy∕48Gy) and D(99) by 12.3% ± 21.6% and 7.3 ± 5.3 Gy, the normal tissues doses by 0.5 Gy on average, gEUD and TCP by 13.0 ± 8.6 Gy and 7.1% ± 5.1%. Comparing 4D(MC) to 3D(MC) doses, V(60Gy∕48Gy) and D(99) of GTV was reduced by 5.2% ± 8.8% and 2.6 ± 3.3 Gy, and normal tissues hardly changed from 4D(MC) to 3D(MC) doses. The corresponding decreases of gEUD and TCP were 2.8 ± 4.0 Gy and 1.6 ± 2.4%. CONCLUSIONS: The large discrepancy between original 3D(EPL) plan and benchmarking 4D(MC) plan is predominately due to dose calculation algorithms as the tracking beam motion and organ deformation hardly influenced doses of normal tissues and moderately decreased V(60Gy∕48Gy) and D(99) of GTV. It is worth to make a thoughtful weight of the benefits of full 4D(MC) dose calculation and consider 3D(MC) dose calculation as a compromise of 4D(MC) dose calculation considering the multifold computation time.

Accuracy and sensitivity of four-dimensional dose calculation to systematic motion variability in stereotatic body radiotherapy (SBRT) for lung cancer.

The dynamic movement of radiation beam in real-time tumor tracking may cause overdosing to critical organs surrounding the target. The primary objective of this study was to verify the accuracy of the 4D planning module incorporated in CyberKnife treatment planning system. The secondary objective was to evaluate the error that may occur in the case of a systematic change of motion pattern. Measurements were made using a rigid thorax phantom. Target motion was simulated with two waveforms (sin and cos4) of different amplitude and frequency. Inversely optimized dose distributions were calculated in the CyberKnife treatment planning system using the 4D Monte Carlo dose calculation algorithm. Each plan was delivered to the phantom assuming (1) reproducible target motion,and (2) systematic change of target motion pattern. The accuracy of 4D dose calculation algorithm was assessed using GAFCHROMIC EBT2 films based on 5%/3 mm γ criteria. Treatment plans were considered acceptable if the percentage of pixels passing the 5%/3 mm γ criteria was greater than 90%. The mean percentages of pixels passing were 95% for the target and 91% for the static off-target structure, respectively, with reproducible target motion. When systematic changes of the motion pattern were introduced during treatment delivery, the mean percentages of pixels passing decreased significantly in the off-target films (48%; p < 0.05), but did not change significantly in the target films (92%; p = 0.324) compared to results of reproducible target motion. These results suggest that the accuracy of 4D dose calculation, particularly in off-target stationary structure, is strongly tied to the reproducibility of target motion and that the solutions of 4D planning do not reflect the clinical nature of nonreproducible target motion generally.

Investigation of four-dimensional (4D) Monte Carlo dose calculation in real-time tumor tracking stereotatic body radiotherapy for lung cancers.

PURPOSE: To investigate the dosimetric variations and radiobiological impacts as a consequence of delivering treatment plans of 3D nature in 4D manner based on the 4D Monte Carlo treatment planning framework implemented on Cyberknife. METHODS AND MATERIALS: Dose distributions were optimized on reference 3D images at end of exhale phase of a 4DCT dataset for twenty-five lung cancer patients treated with 60 Gy / 3Fx or 48 Gy / 4Fx. Deformable image registrations (DIR) between individual 3DCT images to the reference 3DCT image in the 4DCT study were performed to interpolate doses calculated on multiple anatomical geometries back on to the reference geometry to compose a 4D dose distribution that included the tracking beam motion and organ deformation. The 3D and 4D dose distributions that were initially calculated with the equivalent path-length (EPL) algorithm (3DEPL dose and 4DEPL dose) were recalculated with the Monte Carlo algorithm (3DMC dose and 4DMC dose). Dosimetric variations of V60Gy / 48Gy and D99 of GTV, mean doses to the lung and the heart and maximum dose (D1 ) of the spinal cord as a consequence of tracking beam motion in deforming anatomy, dose calculation algorithm, and both were quantified by the relative change from 4DMC to 3DMC doses, from 4DMC to 4DEPL doses, and from 4DMC to 3DEPL doses, respectively. RESULTS: Comparing 4DMC to 3DEPL plans, V60Gy / 48Gy and D99 of GTV decreased considerably by 13 ± 22% (mean ± 1SD) and 9.2 ± 5.5 Gy but changes of normal tissue doses were not more than 0.5 Gy on average. The generalized equivalent uniform dose (gEUD) and tumor control probability (TCP) were reduced by 14.3 ± 8.8 Gy and 7.5 ± 5.2%, and normal tissue complication probability (NTCP) for myelopathy and pericarditis were close to zero and NTCP for radiation pneumonitis was reduced by 2.5 ± 4.1%. Comparing 4DMC to 4DEPL plans found decreased V60Gy / 48Gy and D99 by 12.3 ± 21.6% and 7.3 ± 5.3 Gy, the normal tissues doses by 0.5 Gy on average, gEUD and TCP by 13.0 ± 8.6 Gy and 7.1 ± 5.1%. Comparing 4DMC to 3DMC doses, V60Gy / 48Gy and D99 of GTV was reduced by 5.2 ± 8.8 %and 2.6 ± 3.3 Gy, and normal tissues hardly changed from 4DMC to 3DMC doses. The corresponding decreases of gEUD and TCP were 2.8 ± 4.0 Gy and 1.6 ± 2.4%. CONCLUSION: The large discrepancy between original 3DEPL plan and benchmarking 4DMC plan is predominately due to dose calculation algorithms as the tracking beam motion and organ deformation hardly influenced doses of normal tissues and moderately decreased V60Gy / 48Gy and D99 of GTV. It is worth to make a thoughtful weight of the benefits of full 4D MC dose calculation and consider 3D MC dose calculation as a compromise of 4D MC dose calculation considering the multifold computation time. This article is protected by copyright. All rights reserved.

Validation of a prognostic scoring system for locally recurrent nasopharyngeal carcinoma treated by stereotactic radiosurgery.

BACKGROUND: Selection of patients with local failure of nasopharyngeal carcinoma (NPC) for appropriate type of salvage treatment can be difficult due to the lack of data on comparative efficacy of different salvage treatments. The purpose of the present study was to validate a previously published prognostic scoring system for local failures of NPC treated by radiosurgery based on reported results in the literature. METHODS: A literature search yielded 3 published reports on the use of radiosurgery as salvage treatment of NPC that contained sufficient clinical information for validation of the scoring system. Prognostic scores of 18 patients from these reports were calculated and actuarial survival rates were estimated and compared to the original cohort used to design the prognostic scoring system. The area under the receiver operating characteristic curve was also determined and compared between the current and original patient groups. RESULTS: The calculated prognostic scores ranged from 0.32 to 1.21, with 15 patients assigned to the poor prognostic group and 3 to the intermediate prognostic group. The actuarial 3-year survival rates in the intermediate and poor prognostic groups were 67% and 0%, respectively. These results were comparable to the observed 3-year survival rates of 74% and 23% in the intermediate and poor prognostic group in the original reports. The area under the receiver operating characteristic curve for the current patient group was 0.846 which was similar to 0.841 in the original group. CONCLUSION: The previously published prognostic scoring system demonstrated good prediction of treatment outcome after radiosurgery in a small group of NPC patients with poor prognosis. Prospective study to validate the scoring system is currently being carried out in our institution.

Experimental evaluations of the accuracy of 3D and 4D planning in robotic tracking stereotactic body radiotherapy for lung cancers.

PURPOSE: Due to the complexity of 4D target tracking radiotherapy, the accuracy of this treatment strategy should be experimentally validated against established standard 3D technique. This work compared the accuracy of 3D and 4D dose calculations in respiration tracking stereotactic body radiotherapy (SBRT). METHODS: Using the 4D planning module of the CyberKnife treatment planning system, treatment plans for a moving target and a static off-target cord structure were created on different four-dimensional computed tomography (4D-CT) datasets of a thorax phantom moving in different ranges. The 4D planning system used B-splines deformable image registrations (DIR) to accumulate dose distributions calculated on different breathing geometries, each corresponding to a static 3D-CT image of the 4D-CT dataset, onto a reference image to compose a 4D dose distribution. For each motion, 4D optimization was performed to generate a 4D treatment plan of the moving target. For comparison with standard 3D planning, each 4D plan was copied to the reference end-exhale images and a standard 3D dose calculation was followed. Treatment plans of the off-target structure were first obtained by standard 3D optimization on the end-exhale images. Subsequently, they were applied to recalculate the 4D dose distributions using DIRs. All dose distributions that were initially obtained using the ray-tracing algorithm with equivalent path-length heterogeneity correction (3D EPL and 4D EPL) were recalculated by a Monte Carlo algorithm (3D MC and 4D MC) to further investigate the effects of dose calculation algorithms. The calculated 3D EPL, 3D MC, 4D EPL, and 4D MC dose distributions were compared to measurements by Gafchromic EBT2 films in the axial and coronal planes of the moving target object, and the coronal plane for the static off-target object based on the γ metric at 5%/3mm criteria (γ5%/3mm). Treatment plans were considered acceptable if the percentage of pixels passing γ5%/3mm (Pγ<1) ≥ 90%. RESULTS: The averaged Pγ<1 values of the 3D EPL, 3D MC, 4D EPL, and 4D MC dose calculation methods for the moving target plans are 95%, 95%, 94%, and 95% for reproducible motion, and 95%, 96%, 94%, and 93% for nonreproducible motion during actual treatment delivery. The overall measured target dose distributions are in better agreement with the 3DMC dose distributions than the 4DMC dose distributions. Conversely, measured dose distributions agree much better with the 4D EPL/MC than the 3D EPL/MC dose distributions in the static off-target structure, resulting in higher Pγ<1 values with 4D EPL/MC (91%) vs 3D EPL (24%) and 3D MC (25%). Systematic changes of target motion reduced the averaged Pγ<1 to 47% and 53% for 4D EPL and 4D MC dose calculations, and 22% for 3D EPL/MC dose calculations in the off-target films. CONCLUSIONS: In robotic tracking SBRT, 4D treatment planning was found to yield better prediction of the dose distributions in the off-target structure, but not necessarily in the moving target, compared to standard 3D treatment planning, for reproducible and nonreproducible target motion. It is important to ensure on a patient-by-patient basis that the cumulative uncertainty associated with the 4D-CT artifacts, deformable image registration, and motion variability is significantly smaller than the cumulative uncertainty occurred in standard 3D planning in order to make 4D planning a justified option.

Can intensity-modulated radiotherapy preserve oral health-related quality of life of nasopharyngeal carcinoma patients?

PURPOSE: To investigate the changes in salivary function and oral health-related quality of life for patients with nasopharyngeal carcinoma treated by intensity-modulated radiotherapy (IMRT). METHODS AND MATERIALS: A total of 57 patients with early-stage nasopharyngeal carcinoma received IMRT. The parotid and whole saliva flow was measured, and the Medical Outcomes Study 36-item short form, European Organization for Research and Treatment of Cancer Quality of Life questionnaire-C30, European Organization for Research and Treatment of Cancer Quality of Life questionnaire 35-item head-and-neck module, and Oral Health Impact Profile questionnaires were completed at baseline and 2, 6, 12, 18, and 24 months after IMRT. RESULTS: Parotid saliva flow recovered fully after 1 year and maintained. Whole saliva flow recovered partially to 40% of baseline. A general trend of deterioration in most quality of life scales was observed after IMRT, followed by gradual recovery. Persistent oral-related symptoms were found 2 years after treatment. CONCLUSION: IMRT for early-stage nasopharyngeal carcinoma could only partially preserve the whole salivary function and oral health-related quality of life.

Dosimetric predictors of radiation-induced acute nausea and vomiting in IMRT for nasopharyngeal cancer.

PURPOSE: We wanted to investigate dosimetric parameters that would predict radiation-induced acute nausea and vomiting in intensity-modulated radiation therapy (IMRT) for undifferentiated carcinoma of the nasopharynx (NPC). METHODS AND MATERIALS: Forty-nine consecutive patients with newly diagnosed NPC were treated with IMRT alone in this prospective study. Patients receiving any form of chemotherapy were excluded. The dorsal vagal complex (DVC) as well as the left and right vestibules (VB-L and VB-R, respectively) were contoured on planning computed tomography images. A structure combining both the VB-L and the VB-R, named VB-T, was also generated. All structures were labeled organs at risk (OAR). A 3-mm three-dimensional margin was added to these structures and labeled DVC+3 mm, VB-L+3 mm, VB-R+3 mm, and VB-T+3 mm to account for physiological body motion and setup error. No weightings were given to these structures during optimization in treatment planning. Dosimetric parameters were recorded from dose-volume histograms. Statistical analysis of parameters' association with nausea and vomiting was performed using univariate and multivariate logistic regression. RESULTS: Six patients (12.2%) reported Grade 1 nausea, and 8 patients (16.3%) reported Grade 2 nausea. Also, 4 patients (8.2%) complained of Grade 1 vomiting, and 4 patients (8.2%) experienced Grade 2 vomiting. No patients developed protracted nausea and vomiting after completion of IMRT. For radiation-induced acute nausea, V40 (percentage volume receiving at least 40Gy) to the VB-T and V40>=80% to the VB-T were predictors, using univariate analysis. On multivariate analysis, V40>=80% to the VB-T was the only predictor. There were no predictors of radiation-induced acute vomiting, as the number of events was too small for analysis. CONCLUSIONS: This is the first study demonstrating that a V40 to the VB-T is predictive of radiation-induced acute nausea. The vestibules should be labeled as sensitive OARs, and weightings should be considered for dose sparing during optimization in the treatment planning of IMRT.

Comparative analysis of dosimetric parameters of three different radiation techniques for patients with Graves' ophthalmopathy treated with retro-orbital irradiation.

BACKGROUND: We would like to investigate the if IMRT produced better target coverage and dose sparing to adjacent normal structures as compared with 3-dimensional conformal radiotherapy (3DCRT) and lateral opposing fields (LOF) for patients with Graves' ophthalmopathy treated with retro-orbital irradiation. METHODS: Ten consecutive patients diagnosed with Graves' ophthalmopathy were prospectively recruited into this study. An individual IMRT, 3DCRT and LOF plan was created for each patient. Conformity index (CI), homogeneity index (HI) and other dosimetric parameters of the targets and organs-at-risk (OAR) generated by IMRT were compared with the other two techniques. RESULTS: Mann-Whitney U test demonstrated that CI generated by IMRT was superior to that produced by 3DCRT and LOF (p=0.005 for both respectively). Similarly HI with IMRT was proven better than 3DCRT (p=0.007) and LOF (p=0.005). IMRT gave rise to better dose sparing to some OARs including globes, lenses and optic nerves as compared with 3DCRT but not with LOF. CONCLUSIONS: IMRT, as compared with 3DCRT and LOF, was found to have a better target coverage, conformity and homogeneity and dose sparing to some surrounding structures, despite a slight increase but clinically negligible dose to other structures. Dosimetrically it might be a preferred treatment technique and a longer follow up is warranted to establish its role in routine clinical use.