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

Effects of fluoxetine on functional outcomes after acute stroke (FOCUS): a pragmatic, double-blind, randomised, controlled trial

Background Results of small trials indicate that fluoxetine might improve functional outcomes after stroke. The FOCUS trial aimed to provide a precise estimate of these effects. Methods FOCUS was a pragmatic, multicentre, parallel group, double-blind, randomised, placebo-controlled trial done at 103 hospitals in the UK. Patients were eligible if they were aged 18 years or older, had a clinical stroke diagnosis, were enrolled and randomly assigned between 2 days and 15 days after onset, and had focal neurological deficits. Patients were randomly allocated fluoxetine 20 mg or matching placebo orally once daily for 6 months via a web-based system by use of a minimisation algorithm. The primary outcome was functional status, measured with the modified Rankin Scale (mRS), at 6 months. Patients, carers, health-care staff, and the trial team were masked to treatment allocation. Functional status was assessed at 6 months and 12 months after randomisation. Patients were analysed according to their treatment allocation. This trial is registered with the ISRCTN registry, number ISRCTN83290762. Findings Between Sept 10, 2012, and March 31, 2017, 3127 patients were recruited. 1564 patients were allocated fluoxetine and 1563 allocated placebo. mRS data at 6 months were available for 1553 (99·3%) patients in each treatment group. The distribution across mRS categories at 6 months was similar in the fluoxetine and placebo groups (common odds ratio adjusted for minimisation variables 0·951 [95% CI 0·839–1·079]; p=0·439). Patients allocated fluoxetine were less likely than those allocated placebo to develop new depression by 6 months (210 [13·43%] patients vs 269 [17·21%]; difference 3·78% [95% CI 1·26–6·30]; p=0·0033), but they had more bone fractures (45 [2·88%] vs 23 [1·47%]; difference 1·41% [95% CI 0·38–2·43]; p=0·0070). There were no significant differences in any other event at 6 or 12 months. Interpretation Fluoxetine 20 mg given daily for 6 months after acute stroke does not seem to improve functional outcomes. Although the treatment reduced the occurrence of depression, it increased the frequency of bone fractures. These results do not support the routine use of fluoxetine either for the prevention of post-stroke depression or to promote recovery of function. Funding UK Stroke Association and NIHR Health Technology Assessment Programme

Electropolishing of Al-7Si-0.3Mg Cast Alloy by Using Perchloric and Nitric Acid Electrolytes

The aim of the present study was to find a good method for electropolishing Al-7Si-0.3Mg cast alloy. Consideration of the fundamental current density-voltage relation and limiting current density has made it possible to identify an optimum applied voltage for good electropolishing through variation of different parameters (electrolyte, bath temperature, electrode distance, surface roughness before polishing, cathode material, etc.). The significant findings of the present study are that (1) using 30% perchloric acid + 20% glycerol moderator + 50% methanol electrolyte, (2) maintaining 15 mm electrode distance and a temperature of $-33^0C$, and (3) electropolishing the Al- Si-0.3Mg cast alloy at 15 V for 2 minutes reveal the microstructures clearly. The limiting current density is about $7 mA/cm^2$ for the preceding conditions for attaining good electropolishing. Electropolishing can also be achieved under the same conditions even at room temperature $(26^0C)$, but the clarity of the microstructure is not as good as that obtained through electropolishing at -33°C

Development of electropolishing techniques on metals and alloys

This paper concerns the optimization of electropolishing techniques which are in use to prepare metallographic specimens for microstructural observation. Process variables such as bath temperature, concentration of electrolyte, electrode distance etc., are discussed. Investigations of the influence of process variables on current density-voltage relationship and limiting current density aimed at obtaining a good polished surface in an easy and systematic way are reviewed

White-light emission from yttrium iron garnet (YIG)

Single-phase phosphors that emit broadband white-light are needed for white-light-emitting diodes (wLEDs) to reach their full potential. However, it is challenging to achieve broad white-light emission from single-phase materials. Consequently, polycrystalline inorganic bulk compounds that emit white-light sans doping are rare. We report on broadband white-light emission from a well-known garnet compound, i.e., yttrium iron garnet (YIG), without activator-ion doping. Upon near-UV excitation at 370 nm, polycrystalline bulk YIG emits broadband white-light with (1931) Commission Internationale de L’Eclairage (CIE) coordinates as (0.28, 0.35) and correlated color temperature (CCT) as 8029 K. Variable excitation wavelengths ranging from 280 to 600 nm enable color-tunable emission as cyan-white-blue-green-yellow-orange-red, including near-white-light emission for a broad range of excitation from 325 to 390 nm. Moreover, a short lifetime (sub-nanosecond) is obtained, which is desirable for LED and other applications. We demonstrated the propriety of YIG as a single-phase converting phosphor for illumination by fabricating prototype wLEDs using commercial InGaN UV-LED chips (λ = 380 nm) for excitation. The CIE coordinates and CCT of prototype wLEDs were obtained as (0.34, 0.37) and 5284 K, respectively. We believe that the reported findings signify the great potential of YIG as a single-phase white-light-emitting phosphor for broadband emission, which offers a new perspective and a viable approach for the development of wLEDs

Characteristics of microbial drug resistance and its correlates in chronic diabetic foot ulcer infections

Free to read While virulence factors and the biofilm-forming capabilities of microbes are the key regulators of the wound healing process, the host immune response may also contribute in the events following wound closure or exacerbation of non-closure. We examined samples from diabetic and non-diabetic foot ulcers/wounds for microbial association and tested the microbes for their antibiotic susceptibility and ability to produce biofilms. A total of 1074 bacterial strains were obtained with staphylococci, Pseudomonas, Citrobacter and enterococci as major colonizers in diabetic samples. Though non-diabetic samples had a similar assemblage, the frequency of occurrence of different groups of bacteria was different. Gram-negative bacteria were found to be more prevalent in the diabetic wound environment while Gram-positive bacteria were predominant in non-diabetic ulcers. A higher frequency of monomicrobial infection was observed in samples from non-diabetic individuals when compared to samples from diabetic patients. The prevalence of different groups of bacteria varied when the samples were stratified according to age and sex of the individuals. Several multidrug-resistant strains were observed among the samples tested and most of these strains produced moderate to high levels of biofilms. The weakened immune response in diabetic individuals and synergism among pathogenic micro-organisms may be the critical factors that determine the delicate balance of the wound healing process

Novel CDK12/13 Inhibitors AU-15506 and AU-16770 Are Potent Anti-Cancer Agents in EGFR Mutant Lung Adenocarcinoma with and without Osimertinib Resistance

Osimertinib is a third-generation epidermal growth factor receptor and tyrosine kinase inhibitor (EGFR-TKI) approved for the treatment of lung adenocarcinoma patients harboring EGFR mutations. However, acquired resistance to this targeted therapy is inevitable, leading to disease relapse within a few years. Therefore, understanding the molecular mechanisms of osimertinib resistance and identifying novel targets to overcome such resistance are unmet needs of cancer patients. Here, we investigated the efficacy of two novel CDK12/13 inhibitors, AU-15506 and AU-16770, in osimertinib-resistant EGFR mutant lung adenocarcinoma cells in culture and xenograft models in vivo. We demonstrate that these drugs, either alone or in combination with osimertinib, are potent inhibitors of osimertinib-resistant as well as -sensitive lung adenocarcinoma cells in culture. Interestingly, only the CDK12/13 inhibitor in combination with osimertinib, although not as monotherapy, suppresses the growth of resistant tumors in xenograft models in vivo. Taken together, the results of this study suggest that inhibition of CDK12/13 in combination with osimertinib has the potential to overcome osimertinib resistance in EGFR mutant lung adenocarcinoma patients

PD-1 derived CA-170 is an oral immune checkpoint inhibitor that exhibits preclinical anti-tumor efficacy

Sasikumar et al. describe the identification and characterization of CA-170, a small molecule inhibitor of PD-L1 and VISTA. They find that CA-170 activates T cells and exhibits anti-tumor efficacy in mouse models. This study highlights the potential of CA-170, which has advanced to human clinical trials, as an anti-cancer drug