• PACIFIC: Overall survival with durvalumab versus placebo after chemoradiotherapy in stage III NSCLC

      Raben, D; Faivre-Finn, Corinne; Spigel, D; Daniel, D; Villegas, A; Vincente, D; Hui, R; Carpeno, J; Murakami, S; Paz-Ares, L; Ozguroglu, M; Kurata, T; Chiappori, A; Lee, K; de Wit, M; Poole, L; Wadsworth, C; Dennis, P; Antonia, S; University of Colorado Cancer Center, Aurora, CO (2018)
    • Transitioning adolescent and young adult cancer care research out of its adolescence.

      Stark, D; Fern, LA; Gibson, F; Hawkins, M; Hough, R; McCabe, Martin; Taylor, R; Department of Cancer Medicine, Leeds, UK (2018)
    • Evaluation of bone health assessment in patients receiving glucocorticoids for acute lymphoblastic leukaemia.

      Lewis, Alexander; Khan, Sahrish; Tomlins, Jo; Higham, Claire E; Department of Endocrinology, Christie Hospital, Manchester (2018)
    • Robust radiotherapy planning.

      Unkelbach, J; Alber, M; Bangert, M; Bokrantz, R; Chan, T; Deasy, J; Fredriksson, A; Gorissen, B; van Herk, Marcel; Liu, W; Mahmoudzadeh, H; Nohadani, O; Siebers, J; Witte, M; Xu, H; Department of Radiation Oncology, University Hospital Zurich, Zurich (2018)
      Motion and uncertainty in radiotherapy is traditionally handled via margins. The clinical target volume (CTV) is expanded to a larger planning target volume (PTV), which is irradiated to the prescribed dose. However, the PTV concept has several limitations, especially in proton therapy. Therefore, robust and probabilistic optimization methods have been developed that directly incorporate motion and uncertainty into treatment plan optimization for intensity modulated radiotherapy (IMRT) and intensity modulated proton therapy (IMPT). Thereby, the explicit definition of a PTV becomes obsolete and treatment plan optimization is directly based on the CTV. Initial work focused on random and systematic setup errors in IMRT. Later, inter-fraction prostate motion and intra-fraction lung motion became a research focus. Over the past ten years, IMPT has emerged as a new application for robust planning methods. In proton therapy, range or setup errors may lead to dose degradation and misalignment of dose contributions from different beams - a problem that cannot generally be addressed by margins. Therefore, IMPT has led to the first implementations of robust planning methods in commercial planning systems, making these methods available for clinical use. This paper first summarizes the limitations of the PTV concept. Subsequently, robust optimization methods are introduced and their applications in IMRT and IMPT planning are reviewed.
    • Urgent need for consensus: international survey of clinical practice exploring use of platinum-etoposide chemotherapy for advanced extra-pulmonary high grade neuroendocrine carcinoma (EP-G3-NEC).

      Lamarca, Angela; Frizziero, Melissa; Barriuso, Jorge; McNamara, Mairéad G; Hubner, Richard A; Valle, Juan W; Department of Medical Oncology, The Christie NHS Foundation Trust, Manchester, M20 4BX, UK (2018)
      BACKGROUND: Platinum-etoposide (PE) chemotherapy (CH) is a globally established combination for extra-pulmonary high grade neuroendocrine carcinoma (EP-G3-NEC); the optimal schedule has not been established. METHODS: An international survey was designed, and completed by clinicians with an expertise in the field to assess consistency in clinical practice. RESULTS: Seventy-five replies were received (June-Nov'17). A minority of physicians (13; 17.6%) did not take Ki-67 or morphology (9; 12.0%) into consideration for selection of CH. Most (72; 96.0%) selected PE-CH as first-line treatment for EP-G3-NEC. CH schedules varied: cisplatin-based (37/71; 52.1%), carboplatin-based (34/71; 47.9%); intravenous etoposide (64/71; 90.1%), oral etoposide (7/71; 9.9%). Choice of second-line CH depended on time to progression on PE-based first-line: if?>?6 months, re-challenge with PE was the preferred choice (34; 45.9%); if?<?6 months, alternative combinations such as fluoropyrimidine/irinotecan (21; 29.2%) or temozolomide/capecitabine (22; 30.6%) were used. CONCLUSION: Significant variation in PE regimen employed exists. Standardising clinical practice would facilitate clinical trial development.
    • A real-world study of cardiac events in > 3700 patients with HER2-positive early breast cancer treated with trastuzumab: final analysis of the OHERA study.

      Lidbrink, E; Chmielowska, E; Otremba, B; Bouhlel, A; Lauer, S; Liste, H; Nuesch, E; Shing, M; Misra, Vivek; Karolinska Institute, Karolinska University Hospital, Stockholm, Sweden (2018)
      PURPOSE: Cardiac dysfunction risk associated with intravenous trastuzumab (H IV) treatment may differ in real-world practice versus randomized trials. We investigated cardiac events in patients with HER2-positive early breast cancer (EBC) treated with H IV as adjuvant therapy in routine practice. METHODS: The observational study of cardiac events in patients with HER2-positive EBC treated with Herceptin (OHERA; NCT01152606) enrolled patients with stage I-IIIb disease eligible for H IV in the adjuvant setting per the European Summary of Product Characteristics (SmPC). Primary outcomes were symptomatic congestive heart failure incidence (CHF; New York Heart Association class II-IV) and cardiac death. Patient visits/assessments were per local practice. RESULTS: 3733 Patients received???1 H IV dose per local practice; 88.9% received H IV for >?300 days (median follow-up: ~ 5 years). Prior to disease recurrence (if any), symptomatic CHF occurred in 106 patients (2.8%); 6 (0.2%) cardiac deaths occurred (5 in patients with cardiac disease history). Median time to symptomatic CHF onset was 5.7 months (95% CI 5.3-6.5); 77/106 (72.6%) patients with symptomatic CHF achieved resolution. CHF incidence was higher in patients???65 years, and those with pre-existing cardiac conditions, hypertension, or left ventricular ejection fraction???55% at baseline. CONCLUSIONS: OHERA is the largest prospective observational study to investigate the cardiac safety of H IV as adjuvant EBC therapy in a real-world setting. Symptomatic CHF and cardiac event incidences were consistent with randomized trials in this setting and baseline risk factors identified in the H IV European SmPC.
    • Radiogenomic predictors of adverse effects following charged particle therapy.

      Morton, L; Ricks-Santi, L; West, Catharine ML; Rosenstein, B; Radiation Epidemiology Branch, Division of Cancer Epidemiology and Genetics, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA (2018)
      Radiogenomics is the study of genomic factors that are associated with response to radiation therapy. In recent years, progress has been made toward identifying genetic risk factors linked with late radiation-induced adverse effects. These advances have been underpinned by the establishment of an international Radiogenomics Consortium with collaborative studies that expand cohort sizes to increase statistical power and efforts to improve methodologic approaches for radiogenomic research. Published studies have predominantly reported the results of research involving patients treated with photons using external beam radiation therapy. These studies demonstrate our ability to pool international cohorts to identify common single nucleotide polymorphisms associated with risk for developing normal tissue toxicities. Progress has also been achieved toward the discovery of genetic variants associated with radiation therapy-related subsequent malignancies. With the increasing use of charged particle therapy (CPT), there is a need to establish cohorts for patients treated with these advanced technology forms of radiation therapy and to create biorepositories with linked clinical data. While some genetic variants are likely to impact toxicity and second malignancy risks for both photons and charged particles, it is plausible that others may be specific to the radiation modality due to differences in their biological effects, including the complexity of DNA damage produced. In recognition that the formation of patient cohorts treated with CPT for radiogenomic studies is a high priority, efforts are underway to establish collaborations involving institutions treating cancer patients with protons and/or carbon ions as well as consortia, including the Proton Collaborative Group, the Particle Therapy Cooperative Group, and the Pediatric Proton Consortium Registry. These important radiogenomic CPT initiatives need to be expanded internationally to build on experience gained from the Radiogenomics Consortium and epidemiologists investigating normal tissue toxicities and second cancer risk.
    • Clinical development of novel drug-radiotherapy combinations.

      Ahmad, S; Crittenden, M; Tran, P; Kluetz, P; Blumenthal, G; Bulbeck, H; Baird, R; Williams, K; Illidge, Timothy M; Hahn, S; Lawrence, T; Spears, P; Walker, A; Sharma, R; Department of Oncology, University of Cambridge (2018)
      Radiotherapy is a fundamental component of treatment for the majority of patients with cancer. In recent decades, technological advances have enabled patients to receive more targeted doses of radiation to the tumour, with sparing of adjacent normal tissues. There had been hope that the era of precision medicine would enhance the combination of radiotherapy with targeted anticancer drugs, however this ambition remains to be realised. In view of this lack of progress, the FDA-AACR-ASTRO Clinical Development of Drug-Radiotherapy Combinations Workshop was held in February 2018 to bring together stakeholders and opinion leaders from academia, clinical radiation oncology, industry, patient advocacy groups and the FDA in order to discuss challenges to introducing new drug-radiotherapy combinations to the clinic. This "Perspectives in Regulatory Science and Policy" article summarises the themes and action points that were discussed. Intelligent trial design is required to increase the number of studies which efficiently meet their primary outcomes; endpoints to be considered include local control, organ preservation and patient-reported outcomes. Novel approaches including immune-oncology or DNA repair inhibitor agents combined with radiotherapy should be prioritised. In this article, we focus on how the regulatory challenges associated with defining a new drug-radiotherapy combination can be overcome in order to improve clinical outcomes for patients with cancer.
    • Gene panel testing for breast cancer should not be used to confirm syndromic gene associations.

      Evans, D Gareth R; Howell, Sacha J; Frayling, IM; Peltonen, J; Manchester Centre for Genomic Medicine, Central Manchester University Hospitals NHS Foundation Trust, Manchester, M13 9WL (2018)
    • A rare presentation of hidradenocarcinoma within the penis.

      Cleaveland, Paul; Srivastava, P; Oliveira, Pedro; Parnham, Arie S; Elliott, Tony; Sangar, Vijay K; Christie NHS Foundation Trust, Wilmslow Road, Manchester, M20 4BX, (2018)
    • Consensus on molecular imaging and theranostics in prostate cancer.

      Fanti, S; Minozzi, S; Antoch, G; Banks, I; Briganti, A; Carrio, I; Chiti, A; Clarke, Noel W; Eiber, M; De, B; Fizazi, K; Gillessen, Silke; Gledhill, S; Haberkorn, U; Herrmann, K; Hicks, R; Lecouvet, F; Montironi, R; Ost, P; O'Sullivan, J; Padhani, A; Schalken, J; Scher, H; Tombal, B; van, Moorselaar, R; Van, P; Vargas, H; Walz, J; Weber, W; Wester, H; Oyen, W; Nuclear Medicine Division, Policlinico S Orsola, University of Bologna, Bologna, Italy (2018)
      Rapid developments in imaging and treatment with radiopharmaceuticals targeting prostate cancer pose issues for the development of guidelines for their appropriate use. To tackle this problem, international experts representing medical oncologists, urologists, radiation oncologists, radiologists, and nuclear medicine specialists convened at the European Association of Nuclear Medicine Focus 1 meeting to deliver a balanced perspective on available data and clinical experience of imaging in prostate cancer, which had been supported by a systematic review of the literature and a modified Delphi process. Relevant conclusions included the following: diphosphonate bone scanning and contrast-enhanced CT are mentioned but rarely recommended for most patients in clinical guidelines; MRI (whole-body or multiparametric) and prostate cancer-targeted PET are frequently suggested, but the specific contexts in which these methods affect practice are not established; sodium fluoride-18 for PET-CT bone scanning is not widely advocated, whereas gallium-68 or fluorine-18 prostate-specific membrane antigen gain acceptance; and, palliative treatment with bone targeting radiopharmaceuticals (rhenium-186, samarium-153, or strontium-89) have largely been replaced by radium-223 on the basis of the survival benefit that was reported in prospective trials, and by other systemic therapies with proven survival benefits. Although the advances in MRI and PET-CT have improved the accuracy of imaging, the effects of these new methods on clinical outcomes remains to be established. Improved communication between imagers and clinicians and more multidisciplinary input in clinical trial design are essential to encourage imaging insights into clinical decision making.
    • Electrochemotherapy - emerging applications technical advances, new indications, combined approaches, and multi-institutional collaboration.

      Campana, L; Edhemovic, I; Soden, D; Perrone, A; Scarpa, M; Campanacci, L; Cemazar, M; Valpione, Sara; Miklavcic, D; Mocellin, S; Sieni, E; Sersa, G; Department of Surgery Oncology and Gastroenterology (DISCOG), University of Padua, Italy (2018)
      The treatment of tumors with electrochemotherapy (ECT) has surged over the past decade. Thanks to the transient cell membrane permeabilization induced by the short electric pulses used by ECT, cancer cells are exposed to otherwise poorly permeant chemotherapy agents, with consequent increased cytotoxicity. The codification of the procedure in 2006 led to a broad diffusion of the procedure, mainly in Europe, and since then, the progressive clinical experience, together with the emerging technologies, have extended the range of its application. Herein, we review the key advances in the ECT field since the European Standard Operating Procedures on ECT (ESOPE) 2006 guidelines and discuss the emerging clinical data on the new ECT indications. First, technical developments have improved ECT equipment, with custom electrode probes and dedicated tools supporting individual treatment planning in anatomically challenging tumors. Second, the feasibility and short-term efficacy of ECT has been established in deep-seated tumors, including bone metastases, liver malignancies, and pancreatic and prostate cancers (long-needle variable electrode geometry ECT), and gastrointestinal tumors (endoscopic ECT). Moreover, pioneering studies indicate lung and brain tumors as suitable future targets. A further advance relates to new combination strategies with immunotherapy, gene electro transfer (GET), calcium EP, and radiotherapy. Finally and fourth, cross-institutional collaborative groups have been established to refine procedural guidelines, promote clinical research, and explore new indications.
    • A core outcome set for clinical trials of chemoradiotherapy interventions for anal cancer (CORMAC): a patient and health-care professional consensus.

      Fish, Rebecca; Sanders, C; Adams, R; Brewer, Julie; Brookes, S; DeNardo, Jill; Kochhar, Rohit; Saunders, Mark P; Sebag-Montefiore, D; Williamson, P; Renehan, Andrew G; Division of Cancer Sciences, School of Medical Sciences, Faculty of Biology, Medicine and Health, University of Manchester, Manchester, UK (2018)
      Chemoradiotherapy is the primary treatment for patients with squamous cell carcinoma of the anus, but variations in the reported outcomes have restricted between-study comparisons. Treatment-related morbidity is considerable; however, no trial has comprehensively quantified long-term side-effects or quality of life. Therefore, we established the first international health-care professional and patient consensus to develop a core outcome set, using the Core Outcome Measures in Effectiveness Trials method. We used the results from our previous systematic review and combined them in this Review with patient interviews to derive a comprehensive list of outcomes, followed by a two-round Delphi survey completed by 149 participants (55 patients and 94 health-care professionals) from 11 countries. The Delphi results were discussed at a consensus meeting of health-care professionals and patients. Agreement was reached on 19 outcomes across four domains: disease activity, survival, toxicity, and life impact. Implementation of the Core Outcome Research Measures in Anal Cancer (CORMAC) set in future trials will serve as a framework to achieve standardisation, facilitate selection of health-area-specific evaluation tools, reduce redundancy of outcome lists, allow between-study comparisons, and ultimately enhance the relevance of trial findings to health-care professionals, trialists, and patients.
    • Advanced dose calculation algorithms in lung cancer radiotherapy: implications for SBRT and locally advanced disease in deep inspiration breath hold.

      Josipovic, M; Persson, G; Rydhog, J; Smulders, B; Thomsen, J; Aznar, Marianne, Camille; Department of Oncology, Section of Radiotherapy, Rigshospitalet, Blegdamsvej 9, 2100 Copenhagen, Denmark; (2018)
      PURPOSE: Evaluating performance of modern dose calculation algorithms in SBRT and locally advanced lung cancer radiotherapy in free breathing (FB) and deep inspiration breath hold (DIBH). METHODS: For 17 patients with early stage and 17 with locally advanced lung cancer, a plan in FB and in DIBH were generated with Anisotropic Analytical Algorithm (AAA). Plans for early stage were 3D-conformal SBRT, 45?Gy in 3 fractions, prescribed to 95% isodose covering 95% of PTV and aiming for 140% dose centrally in the tumour. Locally advanced plans were volumetric modulated arc therapy, 66?Gy in 33 fractions, prescribed to mean PTV dose. Calculation grid size was 1?mm for SBRT and 2.5?mm for locally advanced plans. All plans were recalculated with AcurosXB with same MU as in AAA, for comparison on target coverage and dose to risk organs. RESULTS: Lung volume increased in DIBH, resulting in decreased lung density (6% for early and 13% for locally-advanced group). In SBRT, AAA overestimated mean and near-minimum PTV dose (p-values?<?0.01) compared to AcurosXB, with largest impact in DIBH (differences of up to 11?Gy). These clinically relevant differences may be a combination of small targets and large dose gradients within the PTV. In locally advanced group, AAA overestimated mean GTV, CTV and PTV doses by median less than 0.8?Gy and near-minimum doses by median 0.4-2.7?Gy. No clinically meaningful difference was observed for lung and heart dose metrics between the algorithms, for both FB and DIBH. CONCLUSIONS: AAA overestimated target coverage compared to AcurosXB, especially in DIBH for SBRT.
    • Predictive biomarkers of response for immune checkpoint inhibitors in non-small-cell lung cancer.

      Prelaj, Arsela; Tay, Rebecca; Ferrara, R; Chaput, N; Besse, B; Califano, Raffaele; Department of Medical Oncology, The Christie NHS Foundation Trust, Manchester, UK; (2018)
      Immune checkpoint blockade has been a pivotal development in the management of advanced non-small-cell lung cancer (NSCLC). Although durable antitumour activity and improved survival have been observed in a subset of patients, there is a need for additional predictive biomarkers to improve patient selection and avoid toxicity in potential non-responders. This review will address the use and limitations of tumour programmed death-ligand 1 expression as a predictive biomarker and review emerging biomarker strategies specifically related to NSCLC including genetic alterations (tumour mutation burden, loss and gain activated mutations), tumour-related factors (tumour microenvironment) and factors related to the host immune system. Novel approaches in biomarker detection such as peripheral blood monitoring will also be reviewed.
    • The human in vivo biomolecule corona onto PEGylated Liposomes: a proof-of-concept clinical study.

      Hadjidemetriou, M; McAdam, Sarah; Garner, Grace; Thackeray, Chelsey; Knight, D; Smith, Duncan L; Al-Ahmady, Z; Mazza, M; Rogan, Jane; Clamp, Andrew R; Kostarelos, K; Nanomedicine Lab, Faculty of Biology, Medicine and Health, The University of Manchester, AV Hill Building, Manchester, M13 9PT, UK (2018)
      The self-assembled layered adsorption of proteins onto nanoparticle (NP) surfaces, once in contact with biological fluids, is termed the "protein corona" and it is gradually seen as a determinant factor for the overall biological behavior of NPs. Here, the previously unreported in vivo protein corona formed in human systemic circulation is described. The human-derived protein corona formed onto PEGylated doxorubicin-encapsulated liposomes (Caelyx) is thoroughly characterized following the recovery of liposomes from the blood circulation of ovarian carcinoma patients. In agreement with previous investigations in mice, the in vivo corona is found to be molecularly richer in comparison to its counterpart ex vivo corona. The intravenously infused liposomes are able to scavenge the blood pool and surface-capture low-molecular-weight, low-abundance plasma proteins that cannot be detected by conventional plasma proteomic analysis. This study describes the previously elusive or postulated formation of protein corona around nanoparticles in vivo in humans and illustrates that it can potentially be used as a novel tool to analyze the blood circulation proteome.
    • Improving CEA-targeted CAR-T cell therapy for solid tumours.

      Zheng, W; Kueberuwa, Gray; Cheadle, Eleanor J; Armstrong, Anne C; Hawkins, Robert E; University of Manchester, Department Cancer Sciences, Manchester (2018)
    • Impact of intensified chemotherapy in metastatic pancreatic ductal adenocarcinoma (PDAC) in clinical routine in Europe.

      Javed, M; Beyer, G; Le, N; Vinci, A; Wong, H; Palmer, D; Morgan, Robert David; Lamarca, Angela; Hubner, Richard A; Valle, Juan W; Alam, S; Chowdhury, S; Ma, Y; Archibugi, L; Capurso, G; Maisonneuve, P; Neesse, A; Sund, M; Schober, M; Krug, S; NIHR Liverpool Pancreas Biomedical Research Unit, Institute of Translational Medicine, Royal Liverpool University Hospital, United Kingdom (2018)
      BACKGROUND: Pancreatic ductal adenocarcinoma (PDAC) is associated with poor prognosis. Gemcitabine is the standard chemotherapy for patients with metastatic pancreatic adenocarcinoma (MPA). Randomized clinical trials evaluating intensified chemotherapies including FOLFIRINOX and nab-paclitaxel plus gemcitabine (NAB+GEM) have shown improvement in survival. Here, we have evaluated the efficacy of intensified chemotherapy versus gemcitabine monotherapy in real-life settings across Europe. METHODS: A retrospective multi-center study including 1056 MPA patients, between 2012 and 2015, from nine centers in UK, Germany, Italy, Hungary and the Swedish registry was performed. Follow-up was at least 12 months. Cox proportional Harzards regression was used for uni- and multivariable evaluation of prognostic factors. RESULTS: Of 1056 MPA patients, 1030 (98.7%) were assessable for survival analysis. Gemcitabine monotherapy was the most commonly used regimen (41.3%), compared to FOLFIRINOX (n = 204, 19.3%), NAB+GEM (n = 81, 7.7%) and other gemcitabine- or 5-FU-based regimens (n = 335, 31.7%). The median overall survival (OS) was: FOLFIRINOX 9.9 months (95%CI 8.4-12.6), NAB+GEM 7.9 months (95%CI 6.2-10.0), other combinations 8.5 months (95%CI 7.7-9.3) and gemcitabine monotherapy 4.9 months (95%CI 4.4-5.6). Compared to gemcitabine monotherapy, any combination of chemotherapeutics improved the survival with no significant difference between the intensified regimens. Multivariable analysis showed an association between treatment center, male gender, inoperability at diagnosis and performance status (ECOG 1-3) with poor prognosis. CONCLUSION: Gemcitabine monotherapy was predominantly used in 2012-2015. Intensified chemotherapy improved OS in comparison to gemcitabine monotherapy. In real-life settings, the OS rates of different treatment approaches are lower than shown in randomized phase III trials.
    • Physiological changes after colorectal surgery suggest anastomotic leakage is an early event: a retrospective cohort study.

      Stearns, A; Liccardo, F; Tan, K; Sivrikoz, E; Aziz, Omer; Jenkins, J; Kennedy, R; St Mark's Hospital, Watford Road, Harrow, Middlesex, HA1 3UJSt Mark's Hospital, Watford Road, Harrow, Middlesex (2018)
      INTRODUCTION: Anastomotic leakage (AL) is often identified 7-10 days after colorectal surgery. However, in retrospect, patients may have abnormalities evident much earlier. This study aims to identify the clinical time-point when AL occurs. METHODS: This is a retrospective case-matched cohort-comparison study, assessing patients undergoing left-sided colorectal resection between 2006 and 2015 at a specialist colorectal unit. Patients who developed anastomotic leakage (LEAK) were case-matched to two CONTROL patients by procedure, sex, laparoscopic-modality and diverting stoma. Case-note review allowed collection of basic observation data and blood tests (leukocyte count, C-reactive protein, bilirubin, alanine transaminase, creatinine) up to post-operative day (POD)4. The cohorts were compared, with the main outcome measure being changes in basic observation data. RESULTS: Of 554 patients, 49 patients developed AL. These were matched to 98 CONTROL patients. Notes were available for 105 patients (32 LEAK/ 73 CONTROL). Groups were similar in demographics, tumour or nodal status, pre-operative radiotherapy, intra-operative air-leak integrity, and drain usage. AL was detected clinically at a median of 7.5-days post-operatively. There was a significantly increased heart rate by the evening on POD1 in LEAK patients (82.8�14.2 /min-1 versus 75.1�12.7 /min-1 , p=0.0081) which persisted for the rest of the study. By POD3, there was a significant increase in respiratory rate (18.0�4.2 /min-1 versus 16.5�1.3 /min-1 , p=0.0069) and temperature (37.0�0.4C vs 36.7�0.3C, p=0.0006) in LEAK patients. C-reactive protein was significantly higher in LEAK patients from POD2 (165�95mg.L-1 versus 121�75mg/L-1 , p=0.023). CONCLUSIONS: Physiological and biochemical changes associated with AL occur very early post-operatively, suggesting AL may occur within 36 hours after surgery, despite much later clinical detection.