Search

Background: In recent years, there has been an increase in breast surgery for nonpalpable lesions. Traditionally, wires have been used to guide the resection of nonpalpable breast tumors. Recently, several devices have been commercialized as alternatives to classical wires, such as magnetic seeds (Magseed®). The aim of this study is to describe our initial experience using magnetic seeds (Magseed®) to guide breast-conserving surgery in nonpalpable breast lesions and compare the use of magnetic seed with wires to guide breast-conserving surgery, in terms of clinical and pathological characteristics. Methods: We performed a retrospective study including all breast-conserving surgeries for nonpalpable breast lesions Results: Data from 225 cases was collected, 149 magnetic seeds and 76 wires. The breast lesion was localized in every case. Both cohorts were similar regarding clinical and pathological characteristics. We found significant statistical differences (p Conclusion: In our experience, the use of magnetic seed (Magseed®) is a feasible option to guide breast-conserving surgery of non-palpable lesions and enable us to resect less breast tissue.

PurposeThe processes and mechanisms of action which explain how behavioural interventions for chemotherapy-induced peripheral neuropathy (CIPN) work or not are unclear. We describe a co-design process for developing an evidence and theory-based behavioural intervention to reduce the impact of CIPN symptoms on patients’ quality of life.Methods Guided by the Medical Research Council Framework for developing and evaluating complex interventions, our process of intervention development was informed by: (a) findings of systematic reviews, (b) inductive analysis of thirty-nine hours of observational fieldwork, twelve patient and eleven clinician interviews, (c) deductive analysis using the Common-Sense Self-Regulation Model and (d) fifteen patients and twenty clinicians being actively involved as co-designers. ResultsCIPN perception and coping behaviours (including, for example, self-monitoring of symptoms and engaging in self-management and safety strategies to reduce impact of CIPN symptoms) were highlighted as processes to target when co-designing an intervention. To address these behaviours, a psychoeducational approach was deemed suitable. Four behavioural change techniques informed the co-design of the intervention: (a) information about health consequences, (b) salience of consequences, (c) instruction on how to perform a behaviour and (d) action planning.ConclusionCurrent behavioural interventions for CIPN have rarely involved stakeholders, e.g. patients and clinicians, in their development process. We developed a logic model for documenting the proposed mechanism of action of our co-designed intervention for reducing impact of CIPN symptoms. This model will be refined and tested in a subsequent process evaluation as part of a feasibility trial of the intervention.

The flow of suspended capsules through small channels sets up a complex problem presented in different applications, from red blood cells on hemodynamics to flow in porous media. The flow of a susp...

Simple Summary Early reports during the pandemic stated that cancer patients were at increased risk of severe COVID-19 infection and mortality than non-cancer patients. However, delaying cancer treatment can cause worsening symptoms and shorten life expectancy. This study aimed to assess the outcome of cancer patients at our centre to help inform the optimal way to deliver cancer treatment during the COVID-19 pandemic. This study of over 4000 patients found that the delivery of cancer treatment in the first wave of the pandemic did not have a significantly negative effect on mortality. There were low COVID-19 infection (2%) and death rates (6%). As expected, there was a reduction in cancer treatments mainly in later lines of palliative treatments. This likely reflects patient and clinician choice around those more vulnerable patients. This data can be used to re-assure the continuation of cancer treatment during the pandemic. Abstract Background: This study aimed to assess the outcome of cancer patients undergoing systemic anti-cancer treatment (SACT) at our centre to help inform future clinical decision-making around SACT during the COVID-19 pandemic. Methods: Patients receiving at least one episode of SACT for solid tumours at Guy’s Cancer Centre between 1 March and 31 May 2020 and the same period in 2019 were included in the study. Data were collected on demographics, tumour type/stage, treatment type (chemotherapy, immunotherapy, biological-targeted) and SARS-CoV2 infection. Results: A total of 2120 patients received SACT in 2020, compared to 2449 in 2019 (13% decrease). From 2019 to 2020, there was an increase in stage IV disease (62% vs. 72%), decrease in chemotherapy (42% vs. 34%), increase in immunotherapy (6% vs. 10%), but similar rates of biologically targeted treatments (37% vs. 38%). There was a significant increase in 1st and 2nd line treatments in 2020 (68% vs. 81%; p < 0.0001) and reduction in 3rd and subsequent lines (26% vs. 15%; p = 0.004) compared to 2019. Of the 2020 cohort, 2% patients developed SARS-CoV2 infections. Conclusions: These real-world data from a tertiary Cancer Centre suggest that despite the challenges faced due to the COVID-19 pandemic, SACT was able to be continued without any significant effects on the mortality of solid-tumour patients. There was a low rate (2%) of SARS-CoV-2 infection which is comparable to the 1.4%-point prevalence in our total cancer population.

Background: Early reports in the COVID-19 pandemic suggested higher mortality for cancer patients. The impact of potentially immunosuppressive systemic anti-cancer treatments (SACT) was unknown. This study analysed the delivery of SACT for patients with solid malignancies during the COVID-19 outbreak in 2020 compared to the same period in 2019 to inform future clinical decision-making. Methods: All patients receiving at least one SACT at Guy's comprehensive Cancer Centre during the COVID-19 outbreak for solid tumours (1st March- 31st May 2020) were compared to the same period in 2019. SARS-CoV2 infection was by positive RT-PCR test. Data collected: demographics, tumour type/stage and treatment (chemotherapy, immunotherapy (IO), biological-targeted (BT)). Results: 2125 patients received SACT in 2020, compared to 2450 in 2019 (13% decrease). Demographics were comparable with mean age of 62. 56% females in 2020 vs 54% in 2019, 85% vs 83% in the low socio-economic category, 63% vs 73% PS 0-1;30% vs 29% uro-gynaecological, 27% vs 24% breast and 20% vs 23% GI tumours. In 2020 compared to 2019, there was an increase in metastatic disease (71% vs 62%), decrease in CT (34% vs 42%), increase in IO (10% vs 6%), but similar rates of BT treatments (38% vs 37%). Treatment paradigms were similar in 2020 and 2019: neo/adjuvant (28% vs 29%), radical (4% vs 5%) and palliative (69% vs 67%). Earlier palliative treatments were prioritised in 2020 with significant increase in treatments in 1st-2nd line (72% vs 67%;p=0.02) and reduction in > 3rd line (12% vs 27%;p

Resumen Objetivos Revisar la utilidad a largo plazo del manejo percutaneo de los papilomas de mama mediante la extirpacion con BAV guiada por ecografia. Material y metodo Se realizo un estudio retrospectivo de las 122 lesiones papilares (entre 3-30 mm, media 10 mm) extirpadas con BAV guiada por ecografia en nuestro hospital entre abril de 2010 y abril de 2016, en 102 pacientes con edades comprendidas entre 22 y 85 anos (media 59). Presentaban secrecion uniorificial 74 lesiones en 68 pacientes. El protocolo de seguimiento despues de la BAV fue realizar ecografia en 1-2, 6-8 y 12-14 meses, y posteriormente, de forma anual. Si en los controles se detectaban hallazgos sospechosos de papiloma residual o recidivante se realizaba una BAV de rescate. Resultados El resultado de la BAV en las 122 lesiones fue: 114 papilomas benignos (PB), 6 PB con atipia, 2 PB con carcinoma intraductal. Se realizo seguimiento ecografico (6-72 meses, media 54) en las 120 lesiones benignas (100 pacientes). Se sospecho recaida local en 19 PB (19 pacientes, 13 residual y 6 recidivante) y en 4 PB (3 pacientes, recaida a distancia o papilomatosis multiple). Se realizo una nueva BAV en 18 PB con sospecha de recaida local (18 lesiones) y en un PB con recaida a distancia (2 lesiones). En 15 lesiones el resultado fue de PB, en una, de PB con atipia y en 4, de fibrosis. No hubo falsos negativos (carcinomas) en el seguimiento. En una paciente, un PB con recidiva en el interior del pezon se extirpo con cirugia. Dos pacientes desarrollaron papilomatosis multiple incontable y no se realizo BAV. La secrecion desaparecio en 73 papilomas (67 pacientes). Conclusion El tratamiento percutaneo mediante la extirpacion con BAV guiada por ecografia es efectivo en la mayoria de las pacientes con PB o BP con atipia, y seguro en el seguimiento a largo plazo.

3612 Background: The COVID-19 pandemic has hugely affected the spectrum of cancer care. Worldwide healthcare systems have rapidly reorganized cancer services to ensure patients continue to receive essential care whilst optimizing the use of systemic anti-cancer treatments (SACT) and minimizing exposure to SARS-CoV-2 infection. Our study aimed to identify the outcome of patients with gastrointestinal (GI) cancers in our Cancer Centre during the pandemic compared to the same period in 2019. Methods: Retrospective analysis of all GI patients receiving any SACT at Guy’s Cancer Centre from the 1 March-31 May 2020 and 2019. Demographic data (age, ethnicity, socio-economic status (SES), Performance status, cancer and SACT characteristics (type, intent and treatment-line) were collected during both periods. Also we collated the number of COVID-19 infections confirmed by PCR and severity defined by the WHO classification. Patients with clinical or radiological diagnosis were excluded. Results: 567 patients received SACT in 2019 and 417 patients in 2020 (26.4% less). No differences were observed in the demographic or tumour type characteristics. Commonest cancers in both periods were similar: colorectal (47.1, 47%), oesophago-gastric (29, 27.6%), pancreatic-biliary and NET tumours (23.8, 25.4%). However, there were a higher proportion of patients with advanced disease treated in 2020 (70.3% versus 55.2%). Treatment intent was similar in both years: radical (3.5 vs 3.8%), adjuvant (18.2% vs 17.3%), neoadjuvant (15.3% vs 12.7%) and palliative (63% vs 66.2%). There was an increase in the proportion of patients treated in the palliative first line setting (63.8% in 2020 vs 47.6% in 2019) and a reduction in the proportion of third or more treatment (8.7% versus 16.2%) mainly in the colorectal patients. Of 417 GI patients receiving SACT, 14 (3.35%) were diagnosed with COVID-19 infection. Of these, 64.3% were male, 92.9% were low SES and 35.7% were of Caucasian ethnicity. Colorectal cancer was the commonest (57.1%) tumour-type in the COVID-positive group and 57.1% had advanced disease. All the patients that died from COVID-19 were male. 13 patients were on chemotherapy and 1 was on targeted/biological treatment . None was in immunotherapy (n=4). Only one patient was neutropenic (grade 1). 8 patients (57.1%) had severe infection and there were 3 (21.4%) COVID-related deaths. Conclusions: Our study shows the delivery of SACT through the COVID-19 pandemic is relatively safe with low COVID-related mortality rate. It also reflects how we tailored the delivery of anti-cancer treatments to reduce the possible detrimental myelo-suppressive toxicities that could potentially put GI patients at higher risk of severe SARS-CoV-2 infection. This is crucial data that can inform anti-cancer treatment decision making during the protracted COVID-19 pandemic.

Background: The provision of cancer services has been strongly impacted by the outbreak of SARS-CoV-2. Our Cancer Centre in South-East London treats about 8,800 patients annually (incl. 4,500 new diagnoses) and is one of the largest Comprehensive Cancer Centres in the UK. The first COVID-19 positive cancer patient was reported on 29 Feb 2020. Whilst we are dealing with the second wave of COVID-19, it is important to further evaluate safety of cancer treatments whilst balancing risks of COVID-19 infection and complications. Methods: Using descriptive statistics, we report on the patient/tumour characteristics as well as short-term clinical outcomes of those patients undergoing radical treatment (i.e. systemic anticancer treatment (SACT), surgery, or radiotherapy (RT)) for their cancer during the first wave as to help establish the clinical guidelines for the management of cancer patients in a SARS-CoV-2 epidemic. Results: Between March-July 2020, 1,553 patients underwent surgery, 1,125 received SACT, and 814 had RT. Compared to the same period in 2019, there was a decrease of 28% for surgery, 15% for SACT, and 10% for radiotherapy. Whilst surgery was performed on more male patients (58%), more women received SACT (75%) and RT (58%). The age distribution was similar between treatment arms, with the majority of patients aged 50 to 80 years. The most common tumour types were breast (21%), thoracic (20%), and urological (29%) for surgical treatment; breast (49%), gastrointestinal (18%), and gynaecological (10%) for SACT; and breast (40%), urology (25%), and head & neck (11%) for RT. Within SACT, 36% received combination therapy, 35% received systemic chemotherapy, 23% targeted therapy, 5% immunotherapy, and 2% biological therapy. In terms of oncological outcomes, outcomes were similar to pre-COVID-19 times; with 6 deaths at 30 days ( Citation Format: Beth Russell, Victoria Harris, Harvey Dickinson, Charlotte Moss, Graham Roberts, Kate Haire, Eirini Tsotra, Harris Gousis, Debra Josephs, Deborah Enting, Ailsa Sita-Lumsden, Jose Roca, Vasilliki Michalaera, Mary Lei, Angela Swampillai, Eleanor Sawyer, Daniel Smith, Irene De Francesco, Teresa Guerrero-Urbano, Mieke Van Hemelrijck, Saoirse Dolly, Guy's Cancer Real World Evidence. Radical cancer treatment is safe during COVID-19: The experience of a large London-based Comprehensive Cancer Centre [abstract]. In: Proceedings of the AACR Virtual Meeting: COVID-19 and Cancer; 2021 Feb 3-5. Philadelphia (PA): AACR; Clin Cancer Res 2021;27(6_Suppl):Abstract nr P18.

455 Background: The COVID-19 pandemic has prompted difficult decisions around the use of SACT. These were based on limited early data suggesting cancer patients receiving SACT were at an increased risk of COVID-19 severe infection and death. Our study aim was to identify the COVID-19 infection and mortality rates of GI cancer patients receiving SACT. Methods: All GI patients receiving SACT at Guy’s Cancer Centre between March, and May 31, 2020 were included. Demographic data (age, ethnicity, socio-economic status (SES)) and cancer characteristics (stage, SACT type, intent and treatment-line) were collected. COVID-19 infection was confirmed by PCR and severity defined by the World Health Organisation (WHO) classification. Patients with clinical or radiological diagnosis alone were excluded. Results: Of 417 GI patients receiving SACT during the study period, 345 (82.7%) received chemotherapy (alone or combined with targeted/biological treatment), 68 (16.3%) targeted/biological treatment alone and 4 (1%) immunotherapy. 14 (3.4%) patients were diagnosed with COVID-19, 13 were on chemotherapy and 1 on targeted/biological treatment. Commonest cancers in the COVID-19 positive group were colorectal (57.1%) and hepatobiliary (21.4%), followed by oesophago-gastric (14.3%) and neuroendocrine tumours (7.1%); 57.1% had stage IV disease. 64.3% of the positive patients were male (compared to 57.3% in the COVID-19 negative population), mean age was 57.7 years (63.1 years) and 85.7% had low SES (79.7%). 8 (57.1%) patients had severe infection and there were 3 (21.4%) COVID-19 related deaths. All the patients who died from COVID-19 were male and were receiving palliative chemotherapy. Only one patient was neutropenic (grade 1) when diagnosed with COVID-19. Conclusions: The rate of COVID-19 infection in our population was relatively low (3.4 %). Of the 14 COVID-19 positive patients, 57.1% had severe infection, 21.4% died (compared to 3.7% mortality in the non-infected group) and all but one were on chemotherapy. This prospective data, from a large UK comprehensive Cancer Centre, provides some evidence that continuing SACT through the pandemic is relatively safe. The risk of COVID-19 related infection and death must be off-set against the cancer-related morbidity and mortality associated with treatment delays.

Neuropeptide Y (NPY) is widely found in the nervous system and has a role in numerous physiologic processes. In addition, NPY receptors are expressed in neuroendocrine tumors, breast cancer, prostate cancer, kidney cancer, and some types of sarcomas. Different neuropeptides, particularly α-melanocyte-stimulating hormone (MSH), seem to play a role in the pathogenesis of melanoma.We sought to analyze the expression of NPY in cutaneous melanoma, its association with clinical and histologic features, and its correlation with α-MSH.This was an observational study of the immunohistochemical expression of NPY and α-MSH in tissue samples of cutaneous melanomas, different types of melanocytic nevi, and melanoma metastases diagnosed from 2004 to 2008 in San Jorge Hospital, Huesca, Spain.A total of 184 lesions were studied: 49 primary cutaneous melanomas, 12 melanoma metastases (9 cutaneous and 3 lymphatic), and 123 melanocytic nevi. Immunostaining revealed that levels of NPY and α-MSH were significantly higher in melanomas than in melanocytic nevi (P.001). Melanoma metastases were negative for both neuropeptides. Nodular melanomas showed the highest median percentage of NPY positive cells (75% [20-95]) followed by superficial spreading melanoma (25% [2-92]), whereas lentigo maligna were negative (0% [0-0]). Significant, direct associations between NPY expression and vertical growth (P = .0141) and presence of metastasis (P = .0196) were observed. NPY and α-MSH were positively correlated in cutaneous melanoma (0.49, P.001).The sample size of melanomas was not very large.Our study demonstrates that NPY is significantly expressed in melanomas, especially the nodular type, being associated with invasiveness independently of proliferative markers such as thickness, ulceration, and mitotic index.

In the zebrafish-Mycobacterium marinum model for tuberculosis infection, excess TNF triggers necrosis of infected macrophages which leads to exuberant extracellular bacterial growth and host susceptibility (Tobin et al., 2012; Roca and Ramakrishnan, 2013). It has been shown that in infected macrophages, excess TNF induces production of mitochondrial reactive oxygen species (ROS) which activate two death pathways that converge in macrophage necrosis: cyclophilin D-dependent mitochondrial transition pore and acid sphingomyelinase-dependent ceramide production that leads to lysosomal permeabilization. In exploring the events downstream of lysosomal permeabilization, we found that ceramide acts in a Cathepsin D-dependent manner to induce cell death. In the cytosol Cathepsin D initiates the activation of pro-apoptotic factors including Bid and Bax. We engineered Bax mutants that lack its different functional domains and studied the ability of each mutant protein to regulate apoptosis and necrosis. Then we infected Bax-deficient zebrafish larvae expressing the Bax mutants that were unable to induce apoptosis. We found the BH3 domain, which is required for Bax oligomerization during apoptosis, not to be required for necrosis under excess TNF conditions. Rather, an N-terminal transmembrane helix suggested to be the first portion of Bax in contact with the mitochondrial membrane was found to be required for necrosis. Current studies are focused on understanding post-translational modifications of Bax that determine whether activation of the same protein promotes apoptosis or necrosis.

Background: Loss of the metabolic tumor suppressor, argininosuccinate synthetase (ASS1), the rate-limiting enzyme in arginine biosynthesis, sensitizes mesothelioma and lung carcinoma cells to apoptosis following arginine withdrawal. We have reported treatment with the arginine depletor pegylated arginine deiminase (ADI-PEG 20) in ASS1-negative tumor cells potentiates the cytotoxic effect of pemetrexed, accompanied by suppression of de novo pyrimidine synthesis and the pyrimidine salvage pathway (Allen et al, Cancer Res 2014. We have undertaken a phase I study (NCT02029690) of ADI-PEG 20 combined with first-line pemetrexed and cisplatin chemotherapy in patients (pt) with ASS1-deficient mesothelioma or non-squamous non-small cell lung cancer (NSCLC), primary objective was to recommend a dose for future study (RP2D). Methods: Main inclusion criteria: ≥ 18 years, PS ≤ 1, tumour ASS1 loss (≤ 50% ASS expression by IHC) with adequate organ function and written, informed consent. Exclusion criteria: symptomatic CNS metastases, significant concurrent morbidity, therapeutic anticoagulation, history of seizures, or allergy to trial medication(s). A 3+3+3 phase 1 dose escalation design was used with increasing doses of weekly ADI-PEG 20 (18, 27 and 36 mg/m2 IM) in each of three cohorts plus pemetrexed 500 mg/m2 and cisplatin 75 mg/m2 both given every 3 weeks (1 cycle), maximum 18 weeks. ADI-PEG 20 alone was allowed after 18 weeks if there was stable disease or better. Adverse events were graded using CTCAE v4.03 and dose limiting toxicities (DLT) were defined as: Grade 4 neutropenia (> 7 days), febrile neutropenia, Grade 4 anaemia or thrombocytopenia, other clinically significant Grade 3/4 non haematological toxicity that occurred during cycle 1. Radiological disease response was assessed every 6 weeks (modified RECIST for mesothelioma and RECIST 1.1 for NSCLC) and peripheral blood samples were collected to measure plasma arginine and citrulline levels and antibodies to ADI-PEG 20. Results: 42 pt were screened for tumor ASS1 expression and 15 (36%) were ASS1-deficient. Subsequently, 9 pt were eligible for DLT assessment. Demographics - 6 M:3 F, Age range 62 - 77, NSCLC (4): Mesothelioma (5), Prior therapy EGFRi (1 pt), External beam radiotherapy (2 pt) No DLT were observed at any dose level. Grade ≤ 3, AE related to pemetrexed and cisplatin were (5 pt): nausea (5), fatigue (2) and vomiting, peripheral neuropathy, dry mouth, mouth ulcers, dehydration and neutropenia (1 each). No AE were reported related to ADI PEG 20. Mean cycles of treatment: 9 (range 4 - 14) 1 pt was dose reduced after cisplatin toxicity. Arginine was depleted for ≥3 weeks (3-18 weeks in all treated patients. 7/9 pt had partial response (PR) and 2/9 had stable disease (SD) as best response. Conclusions: The combination was well tolerated, the RP2D is 36mg/m2 ADI-PEG 20 weekly with pemetrexed 500 mg/m2 and cisplatin 75 mg/m2 every 3 weeks. Robust clinical activity has been observed with 78% pt having PR as best response on CT scan, as well as PR and SD in2 pt with sarcomatoid mesothelioma. The tolerability and response / disease control rate suggest that this combination may have clinical utility as first line treatment for these malignancies in ASS1-deficient pt. RP2D expansion cohorts are ongoing for pt with pleural mesothelioma or non-squamous NSCLC. Citation Format: Simon Pacey, James F. Spicer, Pui Ying Chan, Mirela Hategan, Dimitra Repana, Jeremy Peter Steele, Peter Schmid, Gary J R Cook, Monica Diaz, Amanda Johnston, Richard Baird, Adelberto Barba, Ramsay Khadeir, Michael Sheaff, Jose Roca, Teresa Szyszko, John Bomalaski, Peter Wojciech Szlosarek. A phase 1 study in patients with mesothelioma or non small cell lung tumours requiring arginine to assess ADI-PEG 20 with pemetrexed and cisplatin (TRAP study). [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr B23.

Macrophages provide the first line of defense against infections through the recognition of different pathogen-associated molecular patterns (PAMPs). Upon activation, they produce pro-inflammatory cytokines, such as interleukin-1β (IL-1β) and tumor necrosis factor α (TNFα), which regulate not only innate but also adaptive immunity. In the bony fish gilthead seabream, we have shown that TNFα produced by activated macrophages induces the recruitment and activation of professional phagocytic granulocytes when injected intraperitoneally, but it hardly activates these cells in vitro. In this study, we report that seabream TNFα is able to up-regulate the expression of E-selectin in endothelial cells, leading to a concomitant increased adhesion of activated granulocytes and lymphocytes. More importantly, although all fish immune cell types analyzed, i.e. granulocytes, macrophages and lymphocytes, produce the CCL4-like chemokine in response to different PAMPs, only lymphocytes and endothelial cells are able to produce this chemokine in response to TNFα. These results demonstrate that TNFα in fish is mainly involved in the recruitment of leukocytes to the inflammatory foci rather than in their activation and suggest a role for fish lymphocytes in the regulation of early immune responses to infection. This work was supported by the Spanish Ministry of Education and Science (grant BIO2005-05078) and the Sixth Framework Programme of the European Union (grant SSP8-CT-2003-501984).

Interaction between chloranil (CA) and n-butylamine (A) at several temperatures has been studied by u.v.–visible spectroscopy. Kinetic and thermodynamic parameters, as well as a mechanism, have been proposed for this reaction. The results indicate the initial formation of charge-transfer complexes between CA and A, in the presence of an excess of amine, which decomposes to give a final product (D)(disubstituted product of CA). The reaction path is autocatalysed by the presence of amine.

Interaction between chloranil (CA) and n-butylamine (A) at several temperatures has been studied by u.v.–visible spectroscopy. Kinetic and thermodynamic parameters, as well as a mechanism, have been proposed for this reaction. The results indicate the initial formation of charge-transfer complexes between CA and A, in the presence of an excess of amine, which decomposes to give a final product (D)(disubstituted product of CA). The reaction path is autocatalysed by the presence of amine.