ALBERT

Beschreibung: The observational status of the polarisation of the anomalous microwave emission (AME) is reviewed, both for individual compact Galactic regions as well as for the large-scale Galactic emission. There are six Galactic regions with existing polarisation constraints in the relevant range of 10–40 GHz: four dust clouds (Perseus, ρ Ophiuchi, LDN1622, and Pleiades) and two HII regions (LPH96 and the Helix nebula). These constraints are discussed in detail and are complemented by deriving upper limits on the polarisation of the AME for those objects without published WMAP constraints. For the case of large-scale emission, two recent works, based on WMAP data, are reviewed. Currently, the best constraints on the fractional polarisation of the AME, at frequencies near the peak of the emission (i.e., 20–30 GHz), are at the level of % (95.4% confidence level). Finally, we compare these constraints with the predictions of some theoretical AME models and discuss the possible impact of polarised AME on future primordial B-mode experiments.

Beschreibung: Evolution of H1N1 influenza A outbreaks of the past 100 years is interesting and significantly complex and details of H1N1 genetic drift remains unknown. Here we investigated the clinical characteristics and immune cross-reactivity of significant historical H1N1 strains. We infected ferrets with H1N1 strains from 1943, 1947, 1977, 1986, 1999, and 2009 and showed each produced a unique clinical signature. We found significant cross-reactivity between viruses with similar HA sequences. Interestingly, A/FortMonmouth/1/1947 antisera cross-reacted with A/USSR/90/1977 virus, thought to be a 1947 resurfaced virus. Importantly, our immunological data that didn't show cross-reactivity can be extrapolated to failure of past H1N1 influenza vaccines, ie. 1947, 1986 and 2009. Together, our results help to elucidate H1N1 immuno-genetic alterations that occurred in the past 100 years and immune responses caused by H1N1 evolution. This work will facilitate development of future influenza therapeutics and prophylactics such as influenza vaccines. Scientific Reports 3 doi: 10.1038/srep01698

Beschreibung: In the frame of the INTERREG III CISM project, sediment cores were collected at 2 stations in the Gulf of Vlorë to study the plankton resting stage assemblages. A total of 87 morphotypes were identified and produced by Dinophyta, Ciliophora, Rotifera, and Crustacea. In 22 cases, the cyst belonged to a species absent from the plankton of the same period. The most abundant resting stages were those produced by Scrippsiella species (Dinophyta). Some calcareous cysts were identified as fossil species associated with Pleistocene to Pliocene sediment, although they were also found in surface sediments and some of them successfully germinated, thus proving their modern status. Total abundance generally decreased with sediment depth at station 40, while station 45 showed distinct maxima at 3 and 8 cm below the sediment surface. The depth of peak abundance in the sediment varied with species. This paper presents the first study of the plankton resting stages in the Bay of Vlorë. The study confirmed the utility of this type of investigation for a more correct evaluation of species diversity. In addition, the varying distribution with sediment depth suggests that this field could be of some importance in determining the history of species assemblages.

Beschreibung: In the frame of the INTERREG III CISM project, sediment cores were collected at 2 stations in the Gulf of Vlorë to study the plankton resting stage assemblages. A total of 87 morphotypes were identified and produced by Dinophyta, Ciliophora, Rotifera, and Crustacea. In 22 cases, the cyst belonged to a species absent from the plankton of the same period. The most abundant resting stages were those produced by Scrippsiella species (Dinophyta). Some calcareous cysts were identified as fossil species associated with Pleistocene to Pliocene sediment, although they were also found in surface sediments and some of them successfully germinated, thus proving their modern status. Total abundance generally decreased with sediment depth at station 40, while station 45 showed distinct maxima at 3 and 8 cm below the sediment surface. The depth of peak abundance in the sediment varied with species. This paper presents the first study of the plankton resting stages in the Bay of Vlorë. The study confirmed the utility of this type of investigation for a more correct evaluation of species diversity. In addition, the varying distribution with sediment depth suggests that this field could be of some importance in determining the history of species assemblages.

Beschreibung: Introduction The phosphatidylinositol 3-kinase (PI3K) pathway is consistently activated in relapsed/refractory Hodgkin lymphoma (HL), suggesting that TGR-1202, a novel inhibitor of the delta isoform of PI3K (PI3K-δ), in clinical development for patients with hematologic malignancies, might represent an attractive therapeutic option. The anti-CD30 monoclonal antibody Brentuximab Vedotin (BV) conjugated to the microtubule-disrupting agent monomethyl auristatin E (MMAE) has recently been reported to induce an overall response rate of 75% in relapsed/refractory HL, but is associated with limited response duration. Combination therapies aimed at enhancing the anti-tumor activity of BV and reducing its side effects may have significant clinical impact in the treatment of relapsed/refractory HL. The present study was aimed at investigating the activity and mechanism(s) of action of the PI3K-δ inhibitor TGR-1202, in combination with BV in non-clinical models of HL. Methods Three HL cell lines, including L-540, KM-H2 and L-428, were used to test the effects of TGR-1202 alone, BV alone, or the combination of TGR-1202 with BV. Cell cycle effects and cell survival were determined by flow cytometry and Western blotting (WB). Additionally, WB was used to assess modulating effects of TGR-1202 on the PI3K/AKT pathway as well as microtubule interacting proteins. Cyclin B1, p21, and α-tubulin were detected by indirect immunofluorescence microscopy. The activity of TGR-1202 and/or BV on microtubule distribution and polymerization were quantified using a three-dimensional volume rendering technique. Results TGR-1202 and BV used as single agents were able to induce a time- and dose-dependent inhibition of cell proliferation and induction of cell death in all cell lines. Compared to the single agent effects, the combination of TGR-1202 (10 µM) and BV (10 ng/ml) synergistically inhibited the mean (±SEM) growth of L-540, KM-H2, and L-428 cell lines (TGR-1202: 40 ± 4%; BV: 30 ± 2%; TGR-1202/BV: 85 ± 1%). Inhibition of cell proliferation induced by the 2-drug combination was associated with a dramatic G2/M cell cycle arrest. Upon TGR-1202/BV treatment, the mean (±SEM) percentages of cells in G2/M phases were increased by 4-fold (72 ± 3%) as compared to TGR-1202 (18 ± 1%) or BV (18 ± 1%) alone. This finding was paralleled by a 3-fold reduction of cells in S phase (TGR-1202: 25 ± 1%; BV: 23 ± 1%; TGR-1202/BV: 9 ± 1%, mean ± SEM) and a marked Cyclin B1 and p21 overexpression. In comparison to each drug as a single agent, the TGR-1202/BV combination led to a synergistic cell death induction. In fact, upon TGR-1202/BV treatment, mean (±SEM) cell death values detected in L-540, KM-H2, and L-428 cell lines were increased by 3-fold over TGR-1202 or BV alone (TGR-1202: 27 ± 2%; BV: 27 ± 2%; TGR-1202/BV: 75 ± 2%). Analysis of caspase-3 and PARP cleavage and blocking experiments with the pan-caspase inhibitor Z-VAD-FMK revealed a caspase-dependent cell death mechanism. In addition, the antiproliferative and cytotoxic effects of TGR-1202 were associated with a marked time-dependent inhibition of PI3K/Akt pathway and dephosphorylation of GSK-3β, Aurora kinases, and stathmin, suggesting that modulation of molecules associated with microtubule polymerization are critically involved in TGR-1202/BV-triggered cell death. To asses potential effects on microtubule dynamics, HL cells were treated with TGR-1202, BV, or the combination for 24 hours, and the effect on microtubules was determined by α-tubulin staining. Compared with controls, TGR-1202 and BV treatment alone led to a modest loss of microtubules (TGR-1202: 11%; BV: 9%), while the combined TGR-1202/BV treatment resulted in a potent synergistic microtubule disruption (mean values of α-tubulin inhibition of 40%, P ≤.0001), supported by a diffuse stain and irregular microtubule fragments throughout the cytosol. Additionally, TGR-1202/BV was found to interfere with the mitotic spindle integrity which may suggest that the G2/M arrest and cytotoxicity of the combined TGR-1202/BV treatment primarily arises from the inhibition of tubulin polymerization. Conclusions Novel PI3K-δ inhibitor TGR-1202 enhances the anti-tumor activity of BV by increasing drug-induced apoptosis and tubulin disruption in all HL cell lines analyzed in the present study. Our data provides a strong rationale for evaluating TGR-1202 in combination with BV in patients with relapsed/refractory HL. Disclosures: Sportelli: TG Therapeutics, Inc.: Employment, Equity Ownership.

Beschreibung: Introduction Disease relapse and resistance to chemotherapy represent challenging issues for Hodgkin Lymphoma (HL) patients. PI3K/AKT and RAF/MEK/ERK pathways are constitutively activated in the majority of HL patients, thus representing attractive therapeutic targets. Previous results from our phase II study indicate that combining the PI3K/AKT inhibitor perifosine with the RAF/MEK/ERK inhibitor sorafenib can achieve significant clinical responses in relapsed/refractory HL. The present study was therefore aimed at characterizing the in vitro and in vivo activity and mechanism(s) of action of a novel PI3K/ERK dual inhibitor AEZS-136 (Æterna Zentaris GmbH, Germany, EU). Methods Four HL cell lines (L-540, SUP-HD1, KM-H2 and L-428) were used to investigate the in vitro effects of AEZS-136 on cell growth, cell cycle distribution, gene expression profiling (GEP), and apoptosis. Live cell imaging experiments were performed to asses the production of reactive oxygen species (ROS). Western blotting (WB) was used to assess the effects of AEZS-136 on MAPK and PI3K/AKT pathways as well as apoptosis. The antitumor efficacy of AEZS-136 was investigated in vivo in nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice. Results Exposure of L-540, SUP-HD1, KM-H2 and L-428 cell lines to AEZS-136 induced a marked, early and time-dependent dephosphorylation of PI3K/Akt and MAPK pathways that was associated with a significant time and dose-dependent cell growth inhibition [80 ± 3% (mean ±SEM) in the L-540 and SUP-HD1 responsive cell lines] and S phase cell cycle arrest. Indeed, upon AEZS-136 treatment the mean (±SEM) percentages of cells in S phase were reduced by 3-fold (13 ± 1%) as compared to control (33 ± 2%). Significant levels of cell death, as assessed by AnnexinV/PI staining, were only observed in L-540 (62 ± 9 vs 14 ± 3%, P ≤.0001) and SUP-HD1 (46 ± 2% vs 15 ± 2%, P ≤.0001) cell lines and were associated with severe mitochondrial dysfunction (up to 40%, P ≤.001). While no activation of caspase-3 and PARP cleavage were observed in L-540 and SUP-HD1 cells treated with AEZS-136, a potent generation of reactive oxygen species (ROS) was observed upon AEZS-136 treatment (up to 90%, P≤.0001). Pretreating cells with the ROS inhibitor YCG063 strongly inhibited AEZS-136-induced ROS generation, mitochondrial dysfunction and cell death, whereas the pan-caspase inhibitor Z-VADfmk did not. Since ROS generation has been implicated in mediating necroptosis, we tested if blocking programmed necrosis with Necrostatin-1 could prevent AEZS-136-induced cytotoxicity. When L-540 cells were treated with AEZS-136 in the presence of Necrostatin-1, cell death and ROS generation were completely prevented, suggesting that cell death was mechanistically related to necroptosis. Additionally, HL cells responsive to AEZS-136-induced cell death showed a pronounced JNK activation whose inhibition by the JNK inhibitor SP600125 reduced cell death and ROS generation. Furthermore, AEZS-136-increased JNK phosphorylation was inhibited by Necrostatin-1 or YCG063, suggesting that ROS-dependent necroptosis was linked to JNK. Interestingly, GEP analysis of L-540 and SUP-HD1 cell lines, but not KM-H2 and L-428 cells, indicated that AEZS-136 treatment induced upregulation of genes involved in positive regulation of cell death. In addition, in KM-H2 and L-428 cells, AEZS-136 strikingly induced the expression of the immediate early response 3 (IER3). Silencing of IER3 restored sensitivity of KM-H2 and L-428 cells to AEZS-136-induced necroptotic cell death, suggesting that IER3 acts as the signaling molecule that mediated AEZS-136-resistance to oxidative cell death. Finally, in vivo experiments were conducted to investigate the antitumor activity of AEZS-136. Treatment of NOD/SCID mice bearing L540 tumor nodules with increasing dose of AEZS-136 (30 – 60 mg/Kg body weight, PO, 5 days/2 weeks) resulted in a dose-dependent reduction of tumor growth (mean TGI of 70%, P ≤.0001) compared to vehicle-treated controls. No mice experienced any apparent treatment-related toxicity. Conclusions The PI3K/ERK dual inhibitor AEZS-136 demonstrates a potent antitumor activity against HL cell lines by targeting aberrant expression of MAPK and PI3K/Akt pathways. These data support further clinical evaluation of AEZS-136 in refractory/relapsed HL patients. Disclosures: No relevant conflicts of interest to declare.