Averaged Differential Expression for the Discovery of Biomarkers in the Blood of Patients with Prostate Cancer

<div><h3>Background</h3><p>The identification of a blood-based diagnostic marker is a goal in many areas of medicine, including the early diagnosis of prostate cancer. We describe the use of averaged differential display as an efficient mechanism for biomarker discovery in whole blood RNA. The process of averaging reduces the problem of clinical heterogeneity while simultaneously minimizing sample handling.</p> <h3>Methodology/Principal Findings</h3><p>RNA was isolated from the blood of prostate cancer patients and healthy controls. Samples were pooled and subjected to the averaged differential display process. Transcripts present at different levels between patients and controls were purified and sequenced for identification. Transcript levels in the blood of prostate cancer patients and controls were verified by quantitative RT-PCR. Means were compared using a t-test and a receiver-operating curve was generated. The Ring finger protein 19A (RNF19A) transcript was identified as having higher levels in prostate cancer patients compared to healthy men through the averaged differential display process. Quantitative RT-PCR analysis confirmed a more than 2-fold higher level of RNF19A mRNA levels in the blood of patients with prostate cancer than in healthy controls (p = 0.0066). The accuracy of distinguishing cancer patients from healthy men using RNF19A mRNA levels in blood as determined by the area under the receiving operator curve was 0.727.</p> <h3>Conclusions/Significance</h3><p>Averaged differential display offers a simplified approach for the comprehensive screening of body fluids, such as blood, to identify biomarkers in patients with prostate cancer. Furthermore, this proof-of-concept study warrants further analysis of RNF19A as a clinically relevant biomarker for prostate cancer detection.</p> </div

The Single-Phase ProtoDUNE Technical Design Report

ProtoDUNE-SP is the single-phase DUNE Far Detector prototype that is under construction and will be operated at the CERN Neutrino Platform (NP) starting in 2018. ProtoDUNE-SP, a crucial part of the DUNE effort towards the construction of the first DUNE 10-kt fiducial mass far detector module (17 kt total LAr mass), is a significant experiment in its own right. With a total liquid argon (LAr) mass of 0.77 kt, it represents the largest monolithic single-phase LArTPC detector to be built to date. It's technical design is given in this report

31st Annual Meeting and Associated Programs of the Society for Immunotherapy of Cancer (SITC 2016) : part two

Background The immunological escape of tumors represents one of the main ob- stacles to the treatment of malignancies. The blockade of PD-1 or CTLA-4 receptors represented a milestone in the history of immunotherapy. However, immune checkpoint inhibitors seem to be effective in specific cohorts of patients. It has been proposed that their efficacy relies on the presence of an immunological response. Thus, we hypothesized that disruption of the PD-L1/PD-1 axis would synergize with our oncolytic vaccine platform PeptiCRAd. Methods We used murine B16OVA in vivo tumor models and flow cytometry analysis to investigate the immunological background. Results First, we found that high-burden B16OVA tumors were refractory to combination immunotherapy. However, with a more aggressive schedule, tumors with a lower burden were more susceptible to the combination of PeptiCRAd and PD-L1 blockade. The therapy signifi- cantly increased the median survival of mice (Fig. 7). Interestingly, the reduced growth of contralaterally injected B16F10 cells sug- gested the presence of a long lasting immunological memory also against non-targeted antigens. Concerning the functional state of tumor infiltrating lymphocytes (TILs), we found that all the immune therapies would enhance the percentage of activated (PD-1pos TIM- 3neg) T lymphocytes and reduce the amount of exhausted (PD-1pos TIM-3pos) cells compared to placebo. As expected, we found that PeptiCRAd monotherapy could increase the number of antigen spe- cific CD8+ T cells compared to other treatments. However, only the combination with PD-L1 blockade could significantly increase the ra- tio between activated and exhausted pentamer positive cells (p= 0.0058), suggesting that by disrupting the PD-1/PD-L1 axis we could decrease the amount of dysfunctional antigen specific T cells. We ob- served that the anatomical location deeply influenced the state of CD4+ and CD8+ T lymphocytes. In fact, TIM-3 expression was in- creased by 2 fold on TILs compared to splenic and lymphoid T cells. In the CD8+ compartment, the expression of PD-1 on the surface seemed to be restricted to the tumor micro-environment, while CD4 + T cells had a high expression of PD-1 also in lymphoid organs. Interestingly, we found that the levels of PD-1 were significantly higher on CD8+ T cells than on CD4+ T cells into the tumor micro- environment (p < 0.0001). Conclusions In conclusion, we demonstrated that the efficacy of immune check- point inhibitors might be strongly enhanced by their combination with cancer vaccines. PeptiCRAd was able to increase the number of antigen-specific T cells and PD-L1 blockade prevented their exhaus- tion, resulting in long-lasting immunological memory and increased median survival

PROPHYLACTIC AND CURATIVE EFFECT OF ETHANOLIC EXTRACT OF BASSIA MALABARICA BARK AGAINST CISPLATIN INDUCED NEPHROTOXICITY

Objective: The present study was designed to investigate the prophylactic and curative effect of ethanolic extract of Bassia malabarica bark (EBBM)against cisplatin (7 mg/Kg single dose i. p) induced nephrotoxicity in male albino rats of Wistar strain.Materials and Methods: After the treatment schedule for 15 days, the extent of nephrotoxicity was quantified using serum samples and isolatedkidneys. Serum samples were used to assess levels of protein, creatinine, urea, uric acid and blood urea nitrogen (BUN). The isolated kidneys wereused to assess antioxidants such as lipid peroxidation (LPO), reduced glutathione (GSH) and catalase (CAT).Results: No mortality was observed in acute toxicity studies conducted as per organization for economic co-operation and development guidelines423 and was found to be safe with no gross behavioral changes up to 2000 mg/Kg body weight (bwt). On administration of cisplatin there was a risein weight of the kidney, creatinine, urea, uric acid, BUN, LPO and a decrease in bwt, urine volume, total protein, GSH and CAT indicating the role ofoxidants to induce nephrotoxicity.Conclusion: Prophylactic and curative groups were found to ameliorate the cisplatin induced alterations in the kidney in a dose-dependent manner.Keywords: Antioxidants, Bassia malabarica, Cisplatin, Nephrocurative, Nephroprotectiv

Role of androgen receptor in progression of LNCaP prostate cancer cells from G1 to S phase.

BACKGROUND: The androgen receptor (AR) plays a critical role in the proliferation of prostate cancer cells. However, its mechanism of action in proliferation remains unknown. An understanding of the mechanism of AR action in proliferation may lead to the development of effective strategies for the treatment of prostate cancer. METHODOLOGY/PRINCIPAL FINDINGS: In this study we report that pulse treatment of synchronized LNCaP cells with Casodex, an AR-antagonist, for 4 hours in mid-G(1) phase was sufficient to prevent cells from entering S phase. Since the assembly of pre-replication complex (pre-RC) in G(1) is required for the progression of cells from G(1) to S phase, the effect of Casodex during mid-G(1) suggested that the role of AR in proliferation might be to regulate the assembly of pre-RC. To test this possibility, we investigated the interaction between AR and Cdc6, an essential component of pre-RC in LNCaP cells. AR co-localized and co-immunoprecipitated with Cdc6, and Casodex treatment disrupted this interaction. AR-immunoprecipitate (AR-IP) also contained cyclin E and cyclin A, which play a critical role in pre-RC assembly and cell cycle entry into S phase, and DNA polymerase-α, PCNA, and ribonucleotide reductase, which are essential for the initiation of DNA synthesis. In addition, in cells in S phase, AR co-sedimented with components of the DNA replication machinery of cells that entered S phase. CONCLUSIONS/SIGNIFICANCE: Together, these observations suggest a novel role of AR as a component of the pre-RC to exert control over progression of LNCaP cells from G(1) to S phase through a mechanism that is independent of its role as a transcription factor

ROC curve evaluating the accuracy of RNF19A as a diagnostic test.

<p>A receiver-operating curve (ROC) was generated as a preliminary estimate of the accuracy of relative levels of RNF19A in classifying patients with cancer or healthy controls in our cohort of patients. The true positive rate (sensitivity) was plotted against the false positive rate (1-specificity). The area under the curve was calculated as 0.7273.</p

ADE analysis of whole blood RNA from prostate cancer patients vs. healthy men.

<p>ADE analysis identified two RNA transcripts with levels significantly higher in whole blood RNA from prostate cancer patients (Ca) than in healthy men (H). Nucleotide sequence analysis revealed that these two transcripts shared a common sequence. This sequence had 100% homology to the nucleotide sequence in the E3-ubiquitin ligase Ring Finger Protein 19a (RNF19A) transcript and is located in the 3′ untranslated region (UTR).</p

Clinical characteristics of the validation prostate cancer cohort.

<p>Characteristics of the 33 patients included in the prostate cancer validation cohort are shown. Age and baseline PSA were abstracted from the time of diagnosis. Gleason score is as recorded by the clinical genitourinary pathologist. Clinical staging of the primary tumor is per the 7^th edition of the American Joint Committee on Cancer (AJCC) staging manual.</p

RNF19A transcript levels are higher in prostate cancer patients than in healthy men.

<p>Quantitative RT-PCR was performed on whole blood RNA samples from patients with localized prostate cancer (n = 33) as well as healthy male controls (n = 19). Levels of RNF19A transcript were normalized to 18S RNA (reference gene). Normalized results are presented in box plot format, with boxes representing the 25^th, 50^th, and 75^th percentiles and whiskers representing the 10^th and 90^th percentiles of the data. Outliers are also displayed. The difference between the means was statistically significant (p  =  0.0066).</p