An integrated approach to determine interactive genotoxic and global gene expression effects of multiwalled carbon nanotubes (MWCNTs) and benzo[a]pyrene (BaP) on marine mussels: evidence of reverse ‘Trojan Horse’ effects

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7q21-rs6964587 and breast cancer risk: an extended case–control study by the Breast Cancer Association Consortium

Background: Using the Breast Cancer Association Consortium, the authors previously reported that the single nucleotide polymorphism 7q21-rs6964587 (AKAP9-M463I) is associated with breast cancer risk. The authors have now assessed this association more comprehensively using 16 independent case–control studies. Methods: The authors genotyped 14 843 invasive case patients and 19 852 control subjects with white European ancestry and 2595 invasive case patients and 2192 control subjects with Asian ancestry. ORs were estimated by logistic regression, adjusted for study. Heterogeneity in ORs was assessed by fitting interaction terms or by subclassifying case patients and applying polytomous logistic regression. Results: For white European women, the minor T allele of 7q21-rs6964587 was associated with breast cancer risk under a recessive model (OR 1.07, 95% CI 1.00 to 1.13, p=0.04). Results were inconclusive for Asian women. From a combined analysis of 24 154 case patients and 33 376 control subjects of white European ancestry from the present and previous series, the best-fitting model was recessive, with an estimated OR of 1.08 (95% CI 1.03 to 1.13, p=0.001). The OR was greater at younger ages (p trend=0.01). Conclusion: This may be the first common susceptibility allele for breast cancer to be identified with a recessive mode of inheritance

Adding 6 months of androgen deprivation therapy to postoperative radiotherapy for prostate cancer: a comparison of short-course versus no androgen deprivation therapy in the RADICALS-HD randomised controlled trial

Background Previous evidence indicates that adjuvant, short-course androgen deprivation therapy (ADT) improves metastasis-free survival when given with primary radiotherapy for intermediate-risk and high-risk localised prostate cancer. However, the value of ADT with postoperative radiotherapy after radical prostatectomy is unclear. Methods RADICALS-HD was an international randomised controlled trial to test the efficacy of ADT used in combination with postoperative radiotherapy for prostate cancer. Key eligibility criteria were indication for radiotherapy after radical prostatectomy for prostate cancer, prostate-specific antigen less than 5 ng/mL, absence of metastatic disease, and written consent. Participants were randomly assigned (1:1) to radiotherapy alone (no ADT) or radiotherapy with 6 months of ADT (short-course ADT), using monthly subcutaneous gonadotropin-releasing hormone analogue injections, daily oral bicalutamide monotherapy 150 mg, or monthly subcutaneous degarelix. Randomisation was done centrally through minimisation with a random element, stratified by Gleason score, positive margins, radiotherapy timing, planned radiotherapy schedule, and planned type of ADT, in a computerised system. The allocated treatment was not masked. The primary outcome measure was metastasis-free survival, defined as distant metastasis arising from prostate cancer or death from any cause. Standard survival analysis methods were used, accounting for randomisation stratification factors. The trial had 80% power with two-sided α of 5% to detect an absolute increase in 10-year metastasis-free survival from 80% to 86% (hazard ratio [HR] 0·67). Analyses followed the intention-to-treat principle. The trial is registered with the ISRCTN registry, ISRCTN40814031, and ClinicalTrials.gov, NCT00541047. Findings Between Nov 22, 2007, and June 29, 2015, 1480 patients (median age 66 years [IQR 61–69]) were randomly assigned to receive no ADT (n=737) or short-course ADT (n=743) in addition to postoperative radiotherapy at 121 centres in Canada, Denmark, Ireland, and the UK. With a median follow-up of 9·0 years (IQR 7·1–10·1), metastasis-free survival events were reported for 268 participants (142 in the no ADT group and 126 in the short-course ADT group; HR 0·886 [95% CI 0·688–1·140], p=0·35). 10-year metastasis-free survival was 79·2% (95% CI 75·4–82·5) in the no ADT group and 80·4% (76·6–83·6) in the short-course ADT group. Toxicity of grade 3 or higher was reported for 121 (17%) of 737 participants in the no ADT group and 100 (14%) of 743 in the short-course ADT group (p=0·15), with no treatment-related deaths. Interpretation Metastatic disease is uncommon following postoperative bed radiotherapy after radical prostatectomy. Adding 6 months of ADT to this radiotherapy did not improve metastasis-free survival compared with no ADT. These findings do not support the use of short-course ADT with postoperative radiotherapy in this patient population

Duration of androgen deprivation therapy with postoperative radiotherapy for prostate cancer: a comparison of long-course versus short-course androgen deprivation therapy in the RADICALS-HD randomised trial

Background Previous evidence supports androgen deprivation therapy (ADT) with primary radiotherapy as initial treatment for intermediate-risk and high-risk localised prostate cancer. However, the use and optimal duration of ADT with postoperative radiotherapy after radical prostatectomy remains uncertain. Methods RADICALS-HD was a randomised controlled trial of ADT duration within the RADICALS protocol. Here, we report on the comparison of short-course versus long-course ADT. Key eligibility criteria were indication for radiotherapy after previous radical prostatectomy for prostate cancer, prostate-specific antigen less than 5 ng/mL, absence of metastatic disease, and written consent. Participants were randomly assigned (1:1) to add 6 months of ADT (short-course ADT) or 24 months of ADT (long-course ADT) to radiotherapy, using subcutaneous gonadotrophin-releasing hormone analogue (monthly in the short-course ADT group and 3-monthly in the long-course ADT group), daily oral bicalutamide monotherapy 150 mg, or monthly subcutaneous degarelix. Randomisation was done centrally through minimisation with a random element, stratified by Gleason score, positive margins, radiotherapy timing, planned radiotherapy schedule, and planned type of ADT, in a computerised system. The allocated treatment was not masked. The primary outcome measure was metastasis-free survival, defined as metastasis arising from prostate cancer or death from any cause. The comparison had more than 80% power with two-sided α of 5% to detect an absolute increase in 10-year metastasis-free survival from 75% to 81% (hazard ratio [HR] 0·72). Standard time-to-event analyses were used. Analyses followed intention-to-treat principle. The trial is registered with the ISRCTN registry, ISRCTN40814031, and ClinicalTrials.gov , NCT00541047 . Findings Between Jan 30, 2008, and July 7, 2015, 1523 patients (median age 65 years, IQR 60–69) were randomly assigned to receive short-course ADT (n=761) or long-course ADT (n=762) in addition to postoperative radiotherapy at 138 centres in Canada, Denmark, Ireland, and the UK. With a median follow-up of 8·9 years (7·0–10·0), 313 metastasis-free survival events were reported overall (174 in the short-course ADT group and 139 in the long-course ADT group; HR 0·773 [95% CI 0·612–0·975]; p=0·029). 10-year metastasis-free survival was 71·9% (95% CI 67·6–75·7) in the short-course ADT group and 78·1% (74·2–81·5) in the long-course ADT group. Toxicity of grade 3 or higher was reported for 105 (14%) of 753 participants in the short-course ADT group and 142 (19%) of 757 participants in the long-course ADT group (p=0·025), with no treatment-related deaths. Interpretation Compared with adding 6 months of ADT, adding 24 months of ADT improved metastasis-free survival in people receiving postoperative radiotherapy. For individuals who can accept the additional duration of adverse effects, long-course ADT should be offered with postoperative radiotherapy. Funding Cancer Research UK, UK Research and Innovation (formerly Medical Research Council), and Canadian Cancer Society

Relative sensitivity of two marine bivalves for detection of genotoxic and cytotoxic effects: a field assessment in the Tamar Estuary, South West England.

The input of anthropogenic contaminants to the aquatic environment is a major concern for scientists, regulators and the public. This is especially relevant in areas such as the Tamar valley in SW England, which has a legacy of contamination from industrial activity in the nineteenth and twentieth centuries. Following on from previous laboratory validation studies, this study aimed to assess the relationship between genotoxic and cytotoxic responses and heavy metal concentrations in two bivalve species sampled from locations along the Tamar estuary. Adult cockles, Cerastoderma edule, and blue mussels, Mytilus edulis, were sampled from five locations in the Tamar and one reference location on the south Devon coast. Bivalve haemocytes were processed for comet and neutral red retention (NRR) assays to determine potential genotoxic and cytotoxic effects, respectively. Sediment and soft tissue samples were analysed for metal content by inductively coupled plasma mass spectrometry. Sediment concentrations were consistent with the physico-chemical nature of the Tamar estuary. A significant correlation (P = 0.05) was found between total metal concentration in sediment and C. edule soft tissues, but no such correlation was found for M. edulis samples. DNA damage was elevated at the site with highest Cr concentrations for M. edulis and at the site with highest Ni and Pb concentrations for C. edule. Analysis of NRR revealed a slight increase in retention time at one site, in contrast to comet data. We conclude that the comet assay is a reliable indicator of genotoxic damage in the field for both M. edulis and C. edule and discuss reasons for the apparent discrepancy with NRR

Combustion of biofuels from acidogenic fermentation in HCCI engines

<p>(A-D) Anti-tubulin immunohistochemical staining (Chromeo™ 488 conjugated secondary antibody) of digestive gland tissue sections from mussels exposed to different experimental conditions (A = Control; B = B[a]P 5 μg/L; C = B[a]P 50 μg/L; D = B[a]P 100 μg/L). (E) Quantitative fluorescence analysis of anti-tubulin immunoreaction. Data are mean ± SD of at least five replicates; * = p < 0.05 (Mann-Whitney <i>U</i>-test).</p

B[a]P-derived-DNA adduct levels, as measured by ³²P-postlabelling, in mussel digestive gland cells, after exposure to 0–1000 μg/L B[a]P for 3 days.

<p>Data are shown with the sexes combined (a) and separated (for those concentrations that resulted in detectable numbers of adducts)(b). All treatments had an n of 9 in total but the distribution of sexes was not even, therefore group n values are indicated in (b). Mismatching lowercase letters indicate differences between groups according to Tukeys HSD applied as post hoc tests following a generalised linear model (p< 0.05; n = 9). Insert: Representative autoradiographic profile of DNA adducts, measured by thin-layer chromatography ³²P-postlabelling, in digestive gland tissue exposed to B[a]P; no B[a]P-derived DNA adducts were detected in control (untreated) tissue (data not shown). The origin (OR), at the bottom left-hand corners, was cut off before exposure. The arrow indicates 10-(deoxyguanosin-<i>N</i>²-yl)-7,8,9-trihydroxy-7,8,9,10-tetrahydro-B[a]P (dG-<i>N</i>²-BPDE).</p

<i>Mytilus galloprovincialis</i> gene expression profiles of digestive gland tissue in animals exposed to B[a]P (5μg/L, 50μg/L and 100 μg/L).

<p>The heat map (A) (Pearson correlation, complete linkage algorithm) reports the log2 relative expression level with respect to the reference condition. 109 differentially expressed genes were generated in at least one condition. Microarray data were analyzed using the Linear Mode for Microarray Analysis (LIMMA) software. B statistics with adjusted p value, 0.05 and B.0 were used as threshold for rejection of the null hypothesis (no variation). Supporting information to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0178460#pone.0178460.g003" target="_blank">Fig 3</a> is presented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0178460#pone.0178460.s005" target="_blank">S2</a> Table and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0178460#pone.0178460.s007" target="_blank">S4 Table</a>. Venn diagram representation of gene expression patterns (Panel B) clearly depicted that no DEGs are shared between the three experimental conditions. All DEGs are obtained respect to the control condition. Data used to generate the Venn-diagram were obtained from microarray analysis (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0178460#pone.0178460.s007" target="_blank">S4 Table</a>). Four biological replications of were considered.</p

Effects of B[a]P on lipofuscin lysosomal content in the digestive gland cells of mussels, evaluated in cryostat tissue sections by the Schmorl reaction.

<p>Data, expressed as percent change with respect to control values, represent the mean ± SD of at least 10 replicates. * = p < 0.05 (Mann-Whitney <i>U</i>-test).</p

Over-representation analysis of DEGs in the digestive gland tissue of mussels exposed to B[a]P (5μg/L, 50μg/L and 100 μg/L).

<p>Showed are: experimental condition; biological processes; Number of up-regulated genes; Number of down-regulated genes. The over-represented biological processes in B[a]P-exposed animals <i>versus</i> control. (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0178460#pone.0178460.s007" target="_blank">S4 Table</a>).</p