Visualising latent DNA on swabs.

Collection for touch DNA either at scenes or on items submitted to a forensic laboratory is based on assumptions as to where a person made direct contact. In many instances a swab may be applied to an area where no contact has been made. Many swabs may therefore be submitted for DNA profiling on which no DNA is present, resulting in the loss of both time and resources by analysing such swabs. This study has developed a simple, fast, DNA-staining and fluorescence microscopy-based screening method for swabs to indicate if there is any DNA from which to generate a profile. Ten different types of swabs were tested covering the major types used (foam, cotton and nylon). Each swab was treated by: no addition of dye or DNA, addition of dye only, addition of known DNA and addition of dye and DNA. The stain used was Diamond™ Nucleic Acid Dye (DD) and fluorescence microscopy was achieved with a digital microscope equipped with a blue LED light source (480nm) for excitation and an emission filter of 510nm. Two types of samples were tested, either buccal swabs or swabs collected from areas touched by volunteers and all analyses were performed in triplicate. The samples were collected and retained at room temperature with time intervals of 0 day, 7 days, 14 days, 21 days, and 28 days before detection using DD staining and fluorescence microscopy. Seven of the swab types used were found to be unsuitable due to the lack of any difference in the fluorescence detected when no DNA, or only the dye, or a combination of DNA and dye were added. Three swab types (black cotton swab, Ultrafine dental applicator, and Cylinder dental applicator) were found to be much more effective for collection of DNA. Further, stained cellular material retained its fluorescence for up to 4 weeks and swabs containing cellular material that had been stored for four weeks could be stained and visualised. Additionally, DD did not affect DNA profiling. This screening method has the potential to be a routine step in a forensic laboratory to save costs of processing samples where swabs are devoid of any DNA. This technique is rapid, easy, cheap, non-destructive and safe.

Direct detection of illicit drugs from biological fluids by desorption/ionization mass spectrometry with nanoporous silicon microparticles.

Surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) is a high throughput analytical technique capable of detecting low molecular weight analytes, including illicit drugs, and with potential applications in forensic toxicology as well as athlete and workplace testing, particularly for biological fluids (oral fluids, urine and blood). However, successful detection of illicit drugs using SALDI-MS often requires extraction steps to reduce the inherent complexity of biological fluids. Here, we demonstrate an all-in-one extraction and analytical system consisting of hydrophobically functionalized porous silicon microparticles (pSi-MPs) for affinity SALDI-MS of prescription and illicit drugs. This novel approach allows for the analysis of drugs from multiple biological fluids without sample preparation protocols. The effect of pSi-MP size, pore diameter, pore depth and functionalization on analytical performance is investigated. pSi-MPs were optimized for the rapid and high sensitivity detection of methadone, cocaine and 3,4-methylenedioxymethamphetamine (MDMA). This optimized system allowed extraction and detection of methadone from spiked saliva and clinical urine samples. Furthermore, by detecting oxycodone in additional clinical saliva and plasma samples, we were able to demonstrate the versatility of the pSi-MP SALDI-MS technique.

Hormone and radiotherapy versus hormone or radiotherapy alone for non-metastatic prostate cancer: a systematic review with meta-analyses.

AIMS: Radiotherapy is standard treatment for localised prostate cancer and is often combined with hormone treatment to prevent androgen stimulation of prostate cancer. Hormone therapy carries significant morbidity and can only be justified in the radical treatment of localised disease if it can be balanced against a significant gain in disease control and survival. MATERIALS AND METHODS: We searched Medline, Premedline, Embase, Cochrane Library, Web of Science (SCI & SSCI) and Biomed Central for randomised controlled trials published in English comparing radiotherapy or hormone therapy alone with radiotherapy and hormone therapy in combination as first-line treatment in patients with non-metastatic prostate cancer reporting overall survival, disease-free survival, distant metastases-free survival, biochemical survival, adverse events (including cardiovascular) and/or health-related quality of life. RESULTS: Fourteen trials were included and showed that combination therapy was associated with better or similar survival and disease-free outcomes compared with single-modality treatment, and that this may particularly be the case for patients with higher risk disease. The results also suggested that combination therapy is associated with more and worse adverse events and quality of life, although this was not always the case. Some of the results are at risk of reporting bias. CONCLUSION: The published data support the use of combined treatment with androgen deprivation and radiotherapy for intermediate- and high-risk localised and locally advanced prostate cancer. Optimal timing, duration, formulation and the management of side-effects remain important questions for further research.

Variations in radiotherapy delivery in England - evidence from the national radiotherapy dataset.

AIMS, MATERIALS AND METHOD: Data in the national radiotherapy dataset for England for 2009-2011 is based upon downloads of activity from every linear accelerator in the country through its oncology management system linked to the local patient administration system to give a full overview of each patient episode. RESULTS: An analysis of this dataset shows that there is still a considerable variation in radiotherapy activity across the country, with a two-fold variation between the most and least active networks. Lower activity is seen in London and the southeast compared with the rest of the country, but when the data are split between the north and south of the country, no such variation is seen. Activity is higher in smaller centres and non-teaching centres. About half of all treatment is palliative and this proportion does not vary with geography, although there is considerable variation between individual centres in the proportion of radical radiotherapy given. There is a trend towards less use of radiotherapy, both radical and palliative, in the more deprived population groups, although no change in the relative use of palliative and radical treatment. CONCLUSION: It is important to emphasise that these data currently reflect activity patterns only and do not reflect quality of care or treatment outcomes, which will be achieved by linkage with cancer registry data in the future.

Endocrine therapy in prostate cancer: time for reappraisal of risks, benefits and cost-effectiveness?

In the 70 years following the first description of the benefits of surgical castration, despite advances in medical therapy e.g. cabazitaxel, enzalutamide, abiraterone, androgen deprivation therapy (ADT) remains the cornerstone of treatment for advanced prostate cancer. However, with increasing numbers of men undergoing PSA testing, the disease is being diagnosed earlier and the costs of ADT, with uncertain survival benefits and associated risks, have risen dramatically. Clinical studies of potent novel agents have shown survival benefits in advanced disease, but timing, risks and cost-effectiveness of treatment remain controversial. As new agents enter clinical practice, a comprehensive research strategy is essential to optimise benefits whilst minimising harm.

The use of lung dose-volume histograms in predicting post-radiation pneumonitis after non-conventionally fractionated radiotherapy for thoracic carcinoma.

AIMS: To assess the use of lung dose-volume histogram (DVH) parameters (specifically V20Gy) in the prediction of radiation pneumonitis for non-conventional fraction sizes used in the treatment of lung cancer. MATERIALS AND METHODS: Patients requiring computed tomography planning for thoracic radiotherapy between January 1999 and January 2002 were identified. The patients receiving radical or high-dose palliative radiotherapy had DVH produced routinely during planning. These were retrospectively reviewed and the case notes accessed for additional pre-treatment parameters, demographics and evidence of radiation pneumonitis. The severity of the pneumonitis was then scored using Radiation Therapy Oncology Group criteria. Data were analysed using the SPSS computer program. RESULTS: One hundred and sixty consecutive patients were reviewed. Ninety patients received hypofractionated treatment (fraction size > 2.5 Gy) and 57 continuous hyperfractionated accelerated radiation therapy (CHART) (fraction size 1.5 Gy). Lung V20Gy values ranged from 3% to 53%, with a median value of 24%. Only six patients reported grade 2, and 16 patients grade 3 pneumonitis. Two patients developed fatal, grade 5 pneumonitis. No correlation between pneumonitis score and V20Gy or other possible predictive factors was found. CONCLUSION: The 15% grade 2-5 pneumonitis rate we document is at the lower end of the spectrum reported in other studies. This suggests that using published data on limiting V20Gy values to reduce the risk of radiation pneumonitis can be extrapolated to planning treatment with non-conventionally fractionated radiotherapy.

A consensus development approach to define national research priorities in bone metastases: proceedings from NCIC CTG workshop.

AIMS: A 1-day workshop was conducted to gather interested Canadian radiation oncologists to identify priority research questions that could be answered through clinical trials under the auspices of the National Cancer Institute of Canada--Clinical Trials Group (NCIC-CTG) Symptom Control committee. MATERIALS AND METHODS: In preparation for the workshop, a survey of Canadian radiation oncologists resulted in four research areas in symptom control, including radiation-induced mucosal reactions, fatigue, radiotherapy for brain metastases and radiotherapy for bone metastases. The first half of the workshop consisted of plenary sessions where the research setting and perspective was defined for each area. This was followed by deliberations by a subgroup of researchers with special interest in the topic area. The bone-metastases subgroup deliberated the clinical context, the scientific merits and the required methodology of research questions related to the role of radiotherapy in early treatment of bone metastases, the role of re-irradiation, the role of systemic radiotherapy and patient selection for different fractionation schedules. A list of prioritised clinical studies was proposed. RESULTS: The question of single vs multi-fraction re-irradiation for symptomatic bone metastases was identified as most pertinent to the Canadian radiation oncologists present. A multi-centre, international intergroup study is undergoing protocol development. Other study concepts, such as an alternative dose-schedule of 17 Gy/2 fractions/1 week for intermediate-prognosis patients, and early referral for radiation oncologist assessment of early or mildly symptomatic bone metastases for good-prognosis patients, require further methodological development before a clinical trial can be proposed. CONCLUSION: An NCIC-CTG workshop provided an update on current evidence-based knowledge in palliative radiotherapy for bone metastases. New trial concepts were discussed among practitioners and clinical investigators to promote dialogue and collaboration. The proposal of an international intergroup randomised trial of single vs multiple fraction re-irradiation for painful bone metastases received the most support among participants.

Calcium distributions in human hair by ToF-SIMS.

Calcium distributions on internal and external surfaces of longitudinally sectioned hairs were analysed with Time-of-Flight Secondary Ion Mass Spectrometry (ToF-SIMS). Externally, calcium deposits were observed at the cuticle scale edges. Internal sections showed that the bulk of calcium exists within or just inside the cuticle layer. The medulla may or may not be enriched and other localised concentrations exist in one of two forms; either associated with granular structures or the hair proteins. Calcium appears to show an affinity for proteins with low sulfur content.

Time-of-flight secondary ion mass spectrometry analysis of hair from archaeological remains.

Hair from four individuals excavated from burial sites in Pacatnamu, Peru from the Moche (450-800 AD) and Lambayeque (900-1100 AD) periods was sectioned longitudinally and analysed with time-of-flight secondary ion mass spectrometry (ToF-SIMS). An attempt was made to distinguish biogenic and diagenetic contributions to the elemental concentrations in the hair samples. Significant contamination was observed to have penetrated the hair samples from the burial environment. Results from the analyses indicate that the burial environment plays an important role in the postmortem variation in elemental content of hair samples. Various elements demonstrated an ability to permeate through the hair matrix over time. In addition, NaCl and what are believed to be aluminosilicates and mineral sulphates, were observed to have accumulated on the surface of the samples. Degradation of the samples was also suspected due to the presence of molecular fragments, possibly resulting from oxidation of the keratin proteins. The results should assist in the identification of reliable elemental signals in the analysis of ancient hair samples and promote caution when considering elements that are abundant in the burial environment.

Primary treatment endpoint following palliative radiotherapy for painful bone metastases: need for a consensus definition?

OBJECTIVES: To compare and contrast the definitions of primary treatment endpoints in randomized studies of dose-fractionation schedules for treating bone metastases and to identify basic characteristics of treatment endpoint that may require consensus among investigators. METHODS: Randomized controlled trials (RCTs) of various dose-fractionation schedules for painful bone metastases, published between 1980 and 1999, and on-going trials whose protocols were available, were systematically reviewed based on the following features of the primary treatment endpoint: (i) degree of pain relief; (ii) timing of the pain response assessment; (iii) effect of co-interventions on pain relief; (iv) the reduction of analgesic as a treatment response; and (v) quantification of response duration. RESULTS: Ten published RCTs (each sampled over 100 patients), plus two current trial protocols were reviewed. Five of the 12 studies defined any reduction in pain score as the primary endpoint. Three trials defined response at pre-determined time points, whereas eight studies attributed pain improvement at any time during follow-up to the effect of radiotherapy. No trial incorporated effect of systemic treatments on response. Only two trials incorporated analgesic scores into the primary endpoint criteria, although several trials reported results of combined pain and analgesic relief. Eight trials reported duration of response. Three provided some estimation of duration with respect to survival: two of them employing actuarial time to pain progression, and one calculated the ratio of pain response to median survival duration (percent net relief). Quality of life was measured in four of 12 studies, as secondary endpoint. CONCLUSION: Although available data suggest similarity in pain relief among various dose-fractionation schedules, accurate and consistent description of the degree of benefit from radiotherapy is lacking. While pain relief is a consistent primary treatment goal among randomized trials, a consensus on several important features of treatment endpoint is needed in order to establish common grounds for future trials in palliative radiotherapy.

Symptom response after palliative radiotherapy for patients with brain metastases.

Whole brain radiotherapy (RT) is frequently used to palliate symptoms in patients with brain metastases, but the palliative benefit to patients has not been well documented. We conducted a longitudinal observational prospective study of patients receiving standard RT (20 Gray (Gy)/5 fractions) for symptomatic brain metastases. End-points were observer rating of neurological symptoms, patient-rated symptoms, performance status, neurological functional status, cognitive function and quality of life (QOL). Median survival for the 75 patients was 86 days (95% confidence interval (CI): 65-101 days). At 1 month, 19% of patients showed an improvement or resolution of presenting symptoms, 23% were stable and 55% had progressed or died. Patient-rated symptoms were increased at 1 month in comparison to baseline data. Only 4 patients had an improved performance status and 22 were stable. Many patients with brain metastases have a short life expectancy and may not benefit from even short duration radiation schedules. Further effort is needed to optimise patient selection and tailor treatment appropriately.

Radiotherapy for brain metastases: defining palliative response.

BACKGROUND AND PURPOSE: Most patients with brain metastases are treated with palliative whole brain radiotherapy (WBRT). There is no established definition of palliative response. The aim of this study was to develop and test clinically useful criteria for response following palliative WBRT. MATERIALS AND METHODS: A prospective study was conducted of patients with symptomatic brain metastases treated with WBRT (20 Gy/5 fractions) and standardised steroid tapering. Assessments included observer rating of neurological symptoms, patient-completed symptom checklist and performance status (PS). Response criteria were operationally defined based on a combination of neurological symptoms, PS and steroid dose. RESULTS: Seventy-five patients were accrued. At 1 month, presenting neurological symptoms were improved in 14 patients, stable in 17, and worse in 21; 23 patients were not assessed, mainly due to death or frailty. Using response criteria defined a priori, 15% (95% CI 7-23%) of patients were classified as having a response to RT, 25% no response, and 29% progression; 27% were deceased at or soon after 1 month. A revised set of criteria was tested, with less emphasis on complete tapering of steroids: they increased the proportion of patients responding to 39% (95% CI 27-50%) but didn't change the large proportion who did not benefit (44%). CONCLUSIONS: Clinical response to RT of patients with brain metastases is multifactorial, comprising symptoms, PS and other factors. Assessment of degree of palliation depend on the exact definition used. More research is needed in this important area, to help validate criteria for assessing palliation after WBRT.

Outcome of curative management of malignant tumours of the parotid gland.

PURPOSE: The optimal management of malignant parotid gland tumours remains to be defined precisely. Specifically, a further understanding of the tumour features that influence treatment outcome is needed. MATERIALS AND METHODS: A retrospective review was conducted on 184 patients who were registered at the Princess Margaret Hospital with a diagnosis of a primary malignant parotid gland tumour. RESULTS: All patients were initially managed with a parotidectomy, and postoperative x-ray radiation therapy (XRT) was administered to 159 patients. The actuarial 5-year cause-specific survival and locoregional control rates were 76% and 81%, respectively. The survival and locoregional control rates for patients treated with surgery alone versus surgery plus postoperative XRT were not statistically different. A multiple regression analysis identified only age and tumour category to be independently significant prognostic factors for both survival and locoregional control. CONCLUSION: We would recommend that patients with malignant parotid gland tumours be managed with parotidectomy, followed by postoperative XRT for tumours with residual disease, aggressive histology, and/or positive lymph nodes.