Vaccination with LAG-3Ig (IMP321) and Peptides Induces Specific CD4 and CD8 T-Cell Responses in Metastatic Melanoma Patients-Report of a Phase I/IIa Clinical Trial.

PURPOSE: Cancer vaccines aim to generate and maintain antitumor immune responses. We designed a phase I/IIa clinical trial to test a vaccine formulation composed of Montanide ISA-51 (Incomplete Freund's Adjuvant), LAG-3Ig (IMP321, a non-Toll like Receptor agonist with adjuvant properties), and five synthetic peptides derived from tumor-associated antigens (four short 9/10-mers targeting CD8 T-cells, and one longer 15-mer targeting CD4 T-cells). Primary endpoints were safety and T-cell responses. EXPERIMENTAL DESIGN: Sixteen metastatic melanoma patients received serial vaccinations. Up to nine injections were subcutaneously administered in three cycles, each with three vaccinations every 3 weeks, with 6 to 14 weeks interval between cycles. Blood samples were collected at baseline, 1-week after the third, sixth and ninth vaccination, and 6 months after the last vaccination. Circulating T-cells were monitored by tetramer staining directly ex vivo, and by combinatorial tetramer and cytokine staining on in vitro stimulated cells. RESULTS: Side effects were mild to moderate, comparable to vaccines with Montanide alone. Specific CD8 T-cell responses to at least one peptide formulated in the vaccine preparation were found in 13 of 16 patients. However, two of the four short peptides of the vaccine formulation did not elicit CD8 T-cell responses. Specific CD4 T-cell responses were found in all 16 patients. CONCLUSIONS: We conclude that vaccination with IMP321 is a promising and safe strategy for inducing sustained immune responses, encouraging further development for cancer vaccines as components of combination therapies. Clin Cancer Res; 22(6); 1330-40. ©2015 AACR

Evaluation of Cavitation Erosion Behavior of Commercial Steel Grades Used in the Design of Fluid Machinery

The erosion response under cavitation of different steel grades was assessed by studying the erosion rate, the volume removal, the roughness evolution, and the accumulated strain energy. A 20 kHz ltrasonic transducer with a probe diameter of 5 mm and peak-to-peak amplitude of 50 lm was deployed in distilled water to induce damage on the surface of commercial chromium and carbon steel samples. After a relatively short incubation period, cavitation induced the formation of pits, cracks, and craters whose features strongly depended on the hardness and composition of the tested steel. AISI 52100 chromium steel showed the best performance and is, therefore, a promising design candidate for replacing the existing fluid machinery materials that operate within potential cavitating environments

Cell cycle, apoptosis, cellular uptake and whole-transcriptome microarray gene expression analysis of HeLa cells treated with a ruthenium(II)-arene complex with an isoquinoline-3-carboxylic acid ligand

Ruthenium(II)-arene complexes are promising drug candidates for the therapy of solid tumors. In previous work, seven new compounds of the general formula [Ru(η6-p-cymene)(L1–7)Cl] were synthesized and characterized, of which the complex with L = isoquinoline-3-carboxylic acid (RuT7) was two times as active on HeLa cells compared to normal cell line MRC-5, as indicated by IC50 values determined after 48 h of incubation (45.4 ± 3.0 vs. 84.2 ± 5.7 μM, respectively). In the present study, cell cycle analysis of HeLa cells treated with RuT7 showed S phase arrest and an increase in sub-G1 population. The apoptotic potential of the title compound was confirmed with the Annexin V-FITC/PI assay together with a morphological evaluation of cells using fluorescent microscopy. Analysis of the intracellular accumulation of ruthenium showed 8.9 ng Ru/106 cells after 6 h of incubation. To gain further insight in the molecular mechanism of action of RuT7 on HeLa cells, a whole-transcriptome microarray gene expression analysis was performed. Analysis of functional categories and signaling and biochemical pathways associated with the response of HeLa cells to treatment with RuT7 showed that it leads the cells through the intrinsic (mitochondrial) apoptotic pathway, via indirect DNA damage due to the action of reactive oxygen species, and through direct DNA binding of RuT7. Statistical analysis for enrichment of gene sets associated with known drug-induced toxicities identified fewer associated toxicity profiles in RuT7-treated cells compared to cisplatin treatment. Altogether these results provide the basis for further development of RuT7 in animal and pre-clinical studies as a potential drug candidate

Spectroscopy and spectropolarimetry of AGN: from observations to modelling

Active galactic nuclei (AGN) are one of the most luminous objects in the Universe, emitting powerful continuum and line emission across all wavelength bands. They represent an important link in the investigations of the galaxy evolution and cosmology. The resolving of the AGN inner structure is still a difficult task with current instruments, therefore the spectroscopy and spectropolarimetry are crucial tools to investigate these objects and their components, such as the properties of the supermassive black hole, the broad line region, and the dusty torus. In this review, we present the results of the project "Astrophysical spectroscopy of extragalactic objects", from the observations, data processing and analysis, to the modelling of different regions in AGN.Comment: Proceedings of the Serbian Astronomical Conference 201

The extremes of AGN variability: outbursts, deep fades, changing looks, exceptional spectral states, and semi-periodicities

The extremes of Active Galactic Nuclei (AGN) variability offer valuable new insights into the drivers and physics of AGN. We discuss some of the most extreme cases of AGN variability; the highest amplitudes, deep minima states, extreme spectral states, Seyfert-type changes, and semi-periodic signals, including new X-ray observations. The properties of changing-look (CL) AGN are briefly reviewed and a classification scheme is proposed which encompasses the variety of CL phenomena; distinguishing slow and fast events, repeat events, and frozen-look AGN which do not show any emission-line response. Long-term light curves that are densely covered over multiple years, along with follow-up spectroscopy, are utilized to gain insight into the underlying variability mechanisms including accretion disk and broad-line region physics. Remarkable differences are seen, for instance, in the optical spectral response to extreme outbursts, implying distinct intrinsic variability mechanisms. Furthermore, we discuss methods for distinguishing between CL AGN and CL look-alike events (tidal disruption events or supernovae in dense media). Finally, semi-periodic light curve variability is addressed and the latest multiwavelength (MWL) light curve of the binary supermassive black hole (SMBH) candidate OJ 287 from the MOMO project is presented. Recent results from that project have clearly established the need for new binary SMBH modelling matching the tight new constraints from observations, including the measurement of a low (primary) SMBH mass of ~10^8 Msun which also implies that OJ 287 is no longer in the regime of near-future pulsar timing arrays

Tumor antigen-specific FOXP3+ CD4 T cells identified in human metastatic melanoma: peptide vaccination results in selective expansion of Th1-like counterparts.

We have previously shown that vaccination of HLA-A2 metastatic melanoma patients with the analogue Melan-A(26-35(A27L)) peptide emulsified in a mineral oil induces ex vivo detectable specific CD8 T cells. These are further enhanced when a TLR9 agonist is codelivered in the same vaccine formulation. Interestingly, the same peptide can be efficiently recognized by HLA-DQ6-restricted CD4 T cells. We used HLA-DQ6 multimers to assess the specific CD4 T-cell response in both healthy individuals and melanoma patients. We report that the majority of melanoma patients carry high frequencies of naturally circulating HLA-DQ6-restricted Melan-A-specific CD4 T cells, a high proportion of which express FOXP3 and proliferate poorly in response to the cognate peptide. Upon vaccination, the relative frequency of multimer+ CD4 T cells did not change significantly. In contrast, we found a marked shift to FOXP3-negative CD4 T cells, accompanied by robust CD4 T-cell proliferation upon in vitro stimulation with cognate peptide. A concomitant reduction in TCR diversity was also observed. This is the first report on direct ex vivo identification of antigen-specific FOXP3+ T cells by multimer labeling in cancer patients and on the direct assessment of the impact of peptide vaccination on immunoregulatory T cells

Comparative cavitation erosion test on steels produced by ESR and AOD refining

Cavitation erosion studies of steels produced by Electroslag Refining (ESR) and Argon Oxygen Decarburization (AOD refining) have been carried out. The experiments were conducted using the modified ultrasonically induced cavitation test method. Erosion rates were measured and the morphology of damages under cavitation action was studied by scanning electron microscopy and optical microscopy techniques. The present work is aimed at understanding the cavitation erosion behaviour of electroslag refined steel (ESR) compared with the steel produced by Argon Oxygen Decarburization (AOD refining), commonly used in the production of hydraulic machinery parts (Pelton blades). The results exhibited lower cavitation rate of ESR steel compared with AOD steel, as a consequence of its better mechanical properties and homogeneous and fine-grained microstructure