The production of volatile compounds from cancer cell lines in vitro has been investigated using selected ion flow tube mass spectrometry (SIFT-MS). This technique enables on-line quantitative analyses of the headspace above cell/medium cultures. This paper reports the discovery that acetaldehyde is released by the lung cancer cell lines SK-MES and CALU-1. The concentration of acetaldehyde in the headspace of the medium/cell culture was measured after 16 h incubation at 37 degrees C and found to be proportional to the number of cancer cells in the medium (typically 10(8)). From these data, the acetaldehyde production rates of the SK-MES cells and the CALU-1 cells in vitro are determined to be 1 x 10(6) and 1.5-3 x 10(6) molecules/cell/min, respectively. The potential value of this new technique in cell biology and in industrial cell biotechnology is discussed.

Previous studies using selected ion flow tube mass spectrometry, SIFT-MS, showed that CALU-1 lung cancer cell cultures emit acetaldehyde in proportion to the number of cells in the culture medium. However, studies in another laboratory led to conflicting results, so these SIFT-MS studies have been repeated and extended to include NL20 normal lung epithelial cells and 35FL121 Tel+ telomerase positive lung fibroblast cells. Thus, SIFT-MS has been used to quantify acetaldehyde and, additionally, carbon dioxide, acetone and ethanol in the headspace of the cell culture medium alone and the headspace of the appropriate medium containing 50 x 10(6) or 80 x 10(6) cells following incubation for 16 h at 37 degrees C. Three independent experiments were carried out for each cell type for both cell numbers and for each of the two culture media used. The results showed that acetone and ethanol were only released by the culture medium, specifically from the foetal calf serum contained in the medium, and not by the cells. Acetaldehyde was also released by the medium, but at relatively lower levels than the other three compounds, and was also generated by the CALU-1 and NL20 cell cultures in proportions to the number of cells in the medium. However, following incubation, the acetaldehyde levels in the headspace of the 35FL121 Tel+ cell cultures were much lower than those present in the headspace of the medium alone. Carbon dioxide was clearly generated by the CALU-1 and 35FL121 Tel+ cells indicating that they were respiring normally, but much less was produced by the NL20 cells, presumably indicating that normal metabolism was being inhibited.

In most industrialized countries, different epidemiologic studies show that chronic renal failure is dramatically increasing. Such major public health problem is a consequence of acquired systemic diseases such as type II diabetes, which is now the first cause for end stage renal failure. Furthermore, lithogenic diseases may also induce intratubular crystallization, which may finally result in end-stage renal failure (ESRF). Up to now, such rare diseases are often misdiagnosed. In this study, based on twenty four biopsies, we show that SR µFTIR (Synchrotron Radiation-µFourier transform infrared) spectroscopy constitutes a significant opportunity to characterize such pathological µcalcifications giving not only their chemical composition but also their spatial distribution in the tissues. This experimental approach offers new opportunities to the clinicians to describe at the cell level the physico-chemical processes leading to the formation of the pathological calcifications which lead to ESRF.

a b s t r a c tIt is widely accepted that health professionals might sometimes underestimate cancer patients' needs for information on the complex process of radiotherapy (RT) planning and delivery. Furthermore, relatives might also feel excluded from the treatment of their loved ones. This pilot study was carried out in order to assess whether both patients and their relatives would welcome further information on RT planning and delivery using the virtual reality (VR) system VERT. One hundred and fifty patients with different types of cancer receiving radical RT were included in the study. Patients and relatives were shown using VERT on a one-to-one basis with an oncologist or a radiographer, a standard room where RT is given, a linear accelerator, and how RT is planned and delivered using their own planning CT Scans. Patients welcomed this information as it helped them to reduce their fears about RT. Relatives felt also more involved in the treatment of their loved one. The results obtained in this pilot study show that VR aids could become an important tool for delivering information on RT to both patients and relatives.© 2015 The College of Radiographers. Published by Elsevier Ltd. All rights reserved. IntroductionThere is clear evidence suggesting there is often a gap between the information provided to patients regarding their disease and management, and the amount of information patients wish to receive.1 Furthermore, it is not uncommon for clinicians to underestimate cancer patients' needs for information, 2 especially when the vast majority of cancer patients want to be thoroughly informed about their illness. 3 Despite this, there are still problems with respect to the provision of information which could satisfy most patients. 4 The importance of the provision of information to patients has been stressed by the Department of Health 5 stating that is a key requirement in current cancer services standards in the UK. However, it cannot be assumed that all patients want substantial information regarding their treatment as some might prefer to fully trust their doctor rather than being involved in taking any decisions that could affect their management. While several studies have addressed these issues, 6,7 further work is still required to improve the understanding of patients' needs. Furthermore, relatives might feel sometimes disengaged in the management of their loved ones and might consider themselves being excluded from several areas within the management process. The negative psychological impact upon partners and other relatives of cancer patients is well documented, 8 and such negative effect is (amongst many other factors) widely associated with lack of medical information given to them. It has long been argued that providing tailored information to relatives as well as patients in general cancer care is a positive aspect of the treatment process 9 and there is little doubt that such inclusionary practice should be encouraged in cancer care.A clear example where provision of information to patien...

Our previous studies have shown that both lung cancer cells and non-malignant lung cells release acetaldehyde in vitro. However, data from other laboratories have produced conflicting results. Furthermore, all these studies have been carried out in 2D models which are less physiological cell growth systems when compared to 3D models. Therefore, we have carried out further work on the release of acetaldehyde by lung cells in 3D collagen hydrogels. Lung cancer cells CALU-1 and non-malignant lung cells NL20 were seeded in these hydrogels at different cell concentrations and the release of acetaldehyde was measured with the Selected Ion Flow Tube Mass Spectrometry (SIFT-MS) technique. The data obtained showed that the amount of acetaldehyde released by both cell types grown in a 3D model is higher when compared to that of the same cells grown in 2D models. More importantly, acetaldehyde from the headspace of lung cancer cells could be measured even at a low cell concentration (10(5) cells per hydrogel). The differential of acetaldehyde release could be, depending on the cell concentration, more than 3 fold higher for cancer cells when compared to non-malignant lung cells. This pilot study is the first to study acetaldehyde emission from albeit only two cell types cultured in 3D scaffolds. Clearly, from such limited data the behaviour of other cell types and of tumour cells in vivo cannot be predicted with confidence. Nevertheless, this work represents another step in the search for volatile biomarkers of tumour cells, the ultimate goal of which is to exploit volatile compounds in exhaled breath and other biological fluids as biomarkers of tumours in vivo.

We have immunized advanced melanoma patients with a HLA-A2-compatible human melanoma line genetically modified to release interleukin-2 (IL-2), to elicit or increase a T cell-mediated anti-melanoma response that may affect distant lesions. Twelve stage-IV patients were injected subcutaneously at days 1, 13, 26, and 55 with IL-2 gene-transduced and irradiated melanoma cells at doses of 5 or 15 x 10(7) cells. Both local and systemic toxicities were mild, consisting of transient erythema at the vaccination site; fever occurred in a minority of patients. Three mixed responses were recorded. Seven patients were evaluable for immunological studies. Mixed tumor-lymphocyte cultures carried out with different allogeneic HLA-A2-matched melanoma lines as stimulators and targets revealed an increase in the MHC-unrestricted, but no changes in the MHC-restricted, cytotoxicity in peripheral blood lymphocytes (PBL) obtained after vaccination as compared with those obtained before vaccination. Increased recognition of the tyrosinase 368-376 peptide occurred in post-vaccination PBL of one patient, whereas a weak increase in recognition of the gp100 280-288 peptide was detectable in another patient; these 2 patients also recognized the gp100 457-466 peptide. After in vitro, stimulation with the only available autologous melanoma line, CD4+ cells with autologous tumor-specific cytotoxicity and ability to release interferon-gamma (IFN-gamma) were found in post- but not in pre-vaccination PBL. In the same patient, as well as in another patient, limiting dilution analysis showed that vaccination resulted in an increased frequency of melanoma-specific cytotoxic T lymphocyte (CTL) precursors. These results indicate that vaccination with cells releasing IL-2 locally can expand a T cell response against antigen(s) of autologous, untransduced tumor, although this response occurred in a minority of the melanoma patients studied.

B7 co-stimulation is necessary to activate resting T cells upon antigen recognition by the T cell receptor. To see whether expression of B7 may render human melanoma cells able to stimulate T cells, a cloned melanoma line (Me1B6), which did not express B7-1, was transfected with the human B7-1 gene. In proliferation assays, B7-1 transfected cells (Me1B6/B7) showed greater stimulatory activity of allogeneic and autologous peripheral blood lymphocytes (PBL) compared to parental, non-transfected tumor cells. This effect was also seen when allogeneic CD8+ and CD4+ subpopulations were used as effectors. In these studies, activation of lymphocytes was B7-1-dependent and HLA classes I and II mediated. The higher proliferation correlated with an increased lytic activity by PBL stimulated with B7-1+ tumor cells against the untransfected Me1B6. Furthermore, PBL from a metastatic melanoma patient stimulated by Me1B6/B7 developed an higher lytic activity not only against Me1B6 but also against their autologous, B7-1- tumor. Finally, after Me1B6/B7 stimulation, PBL released interleukin (IL)-2 and interferon-gamma, but not IL-4, suggesting a Th1-mediated response. These data support the use of B7-1 transfected melanoma cells in the therapeutic vaccination of melanoma patients.

Two human melanoma lines were transduced by a retroviral vector with the gene of the human interleukin-2 (IL-2) and characterized for their immunological properties in comparison with the parental lines. Transduction resulted in the production of biologically active IL-2 in the average amounts of 2,282 and 2,336 pg/ml per 10(5) cells per 24 hr over 3 and 2 months by the Me14932/IL-2 and the Me1B6/IL-2 lines, respectively. Melanoma-transduced cells lost their tumorigenicity in nude mice. No major changes in the phenotype were observed in IL-2 gene-transduced lines. In fact, more than 90% of cells expressed class I and II(DR) HLA, adhesion molecules, integrins, and melanoma-associated antigens. Irradiation with 100-400 Gy, while inhibiting tumor cell growth in vitro, allowed the release of IL-2 by the transduced cells for at least 5 weeks. The two melanoma lines also maintained susceptibility to lysis by lymphokine-activated killer (LAK) cells and by a HLA-A2-restricted melanoma-specific cytotoxic T lymphocyte (CTL) clone recognizing the melanoma antigen (Melan-A). In a limiting dilution assay, transduced, but not parental melanoma lines unless added with an amount of IL-2 comparable to that released by the transduced cells, were able to expand both nonspecific and melanoma-specific CTL precursors from autologous peripheral blood lymphocytes (PBL). In mixed lymphocytes-tumor cultures, IL-2 gene-transduced melanoma cells stimulated the expansion of major histocompatibility complex (MHC)-unrestricted effectors from autologous PBL, and of CD3+ CD8+ MHC-restricted CTL from tumor-invaded lymph nodes. These results indicate that IL-2 gene transduction does not alter significantly the expression of the immunologically relevant molecules of human melanoma lines while increasing their ability to stimulate both specific and nonspecific lymphocyte responses. These lines will be of value in the vaccination of melanoma patients.