The ATR inhibitor elimusertib exhibits anti-lymphoma activity and synergizes with the PI3K inhibitor copanlisib.

The DNA damage response (DDR) is the cellular process of preserving an intact genome and is often deregulated in lymphoma cells. The ataxia telangiectasia and Rad3-related (ATR) kinase is a crucial factor of DDR in the response to DNA single-strand breaks. ATR inhibitors are agents that have shown considerable clinical potential in this context. We characterized the activity of the ATR inhibitor elimusertib (BAY 1895344) in a large panel of lymphoma cell lines. Furthermore, we evaluated its activity combined with the clinically approved PI3K inhibitor copanlisib in vitro and in vivo. Elimusertib exhibits potent anti-tumour activity across various lymphoma subtypes, which is associated with the expression of genes related to replication stress, cell cycle regulation and, as also sustained by CRISPR Cas9 experiments, CDKN2A loss. In several tumour models, elimusertib demonstrated widespread anti-tumour activity stronger than ceralasertib, another ATR inhibitor. This activity is present in both DDR-proficient and DDR-deficient lymphoma models. Furthermore, a combination of ATR and PI3K inhibition by treatment with elimusertib and copanlisib has in vitro and in vivo anti-tumour activity, providing a potential new treatment option for lymphoma patients.

Pharmacologic screen identifies active combinations with BET inhibitors and LRRK2 as a novel putative target in lymphoma.

Inhibitors of the Bromo- and Extra-Terminal domain (BET) family proteins have strong preclinical antitumor activity in multiple tumor models, including lymphomas. Limited single-agent activity has been reported in the clinical setting. Here, we have performed a pharmacological screening to identify compounds that can increase the antitumor activity of BET inhibitors in lymphomas. The germinal center B-cell like diffuse large B-cell lymphoma (DLBCL) cell lines OCI-LY-19 and WSU-DLCL2 were exposed to 348 compounds given as single agents at two different concentrations and in combination with the BET inhibitor birabresib. The combination partners included small molecules targeting important biologic pathways such as PI3K/AKT/MAPK signaling and apoptosis, approved anticancer agents, kinase inhibitors, epigenetic compounds. The screening identified a series of compounds leading to a stronger antiproliferative activity when given in combination than as single agents: the histone deacetylase (HDAC) inhibitors panobinostat and dacinostat, the mTOR (mechanistic target of rapamycin) inhibitor everolimus, the ABL/SRC (ABL proto-oncogene/SRC proto oncogene) inhibitor dasatinib, the AKT1/2/3 inhibitor MK-2206, the JAK2 inhibitor TG101209. The novel finding was the benefit given by the addition of the LRRK2 inhibitor LRRK2-IN-1, which was validated in vitro and in vivo. Genetic silencing demonstrated that LRRK2 sustains the proliferation of lymphoma cells, a finding paired with the association between high expression levels and inferior outcome in DLBCL patients. We identified combinations that can improve the response to BET inhibitors in lymphomas, and LRRK2 as a gene essential for lymphomas and as putative novel target for this type of tumors.

Mechanistic insights into 2,4-D photoelectrocatalytic removal from water with TiO2 nanotubes under dark and solar light irradiation.

Removal of recalcitrant pollutants from water is a major challenge, to which the photoelectrocatalytic processes may be a solution. Applied potential plays a key role in the photocatalytic activity of the semiconductor. This paper investigated the effect of applied potential on the photoelectrocatalytic oxidation of 2,4-Dichlorophenoxyacetic acid (2,4-D) with TiO2 nanotubular anodes under solar light irradiation. The process was investigated at constant potentials in different regions of the polarization curve: the ohmic region, the saturation region and in the region of the Schottky barrier breakdown. PEC tests were performed in aqueous solutions of 2,4-D, and in the presence of methanol or formic acid, as scavengers of OH• radicals and holes. Results showed the main mechanism is oxidation by OH• radicals from water oxidation, while runs with hole scavenger revealed a second mechanism of direct oxidation by holes photogenerated at the electrode surface, with high removal rates due to current doubling effect.

Experimental study on the optimisation of azo-dyes removal by photo-electrochemical oxidation with TiO2 nanotubes.

An experimental investigation is here presented on the photo-electrochemical removal of Methyl Orange (MO), selected as a model of the organic dyes, contained in wastewaters. The process is carried out in an electrochemical flow reactor, in which titania nanotubular electrode is irradiated with a simulated solar light. Design of Experiments (DOE) technique is used to plan the experimental campaign and investigate on the single and combined effects of applied current, electrolyte flow rate, and initial MO concentration, on the specific reaction rate. The results of the DOE analysis, also combined with the study of the distribution of the intermediate products, confirm a reaction mechanism mediated by OH radicals; high applied current and low reactant concentration resulted as favourable conditions to achieve high specific reaction rate of color removal.

Propionate Production by Bioelectrochemically-Assisted Lactate Fermentation and Simultaneous CO2 Recycling.

Production of volatile fatty acids (VFAs), fundamental building blocks for the chemical industry, depends on fossil fuels but organic waste is an emerging alternative substrate. Lactate produced from sugar-containing waste streams can be further processed to VFAs. In this study, electrofermentation (EF) in a two-chamber cell is proposed to enhance propionate production via lactate fermentation. At an initial pH of 5, an applied potential of -1 V vs. Ag/AgCl favored propionate production over butyrate from 20 mM lactate (with respect to non-electrochemical control incubations), due to the pH buffering effect of the cathode electrode, with production rates up to 5.9 mM d-1 (0.44 g L-1 d-1). Microbial community analysis confirmed the enrichment of propionate-producing microorganisms, such as Tyzzerella sp. and Propionibacterium sp. Organisms commonly found in microbial electrosynthesis reactors, such as Desulfovibrio sp. and Acetobacterium sp., were also abundant at the cathode, indicating their involvement in recycling CO2 produced by lactate fermentation into acetate, as confirmed by stoichiometric calculations. Propionate was the main product of lactate fermentation at substrate concentrations up to 150 mM, with a highest production rate of 12.9 mM d-1 (0.96 g L-1 d-1) and a yield of 0.48 mol mol-1 lactate consumed. Furthermore, as high as 81% of the lactate consumed (in terms of carbon) was recovered as soluble product, highlighting the potential for EF application with high-carbon waste streams, such as cheese whey or other food wastes. In summary, EF can be applied to control lactate fermentation toward propionate production and to recycle the resulting CO2 into acetate, increasing the VFA yield and avoiding carbon emissions and addition of chemicals for pH control.

Single and combined BTK and PI3Kδ inhibition with acalabrutinib and ACP-319 in pre-clinical models of aggressive lymphomas.

The B-cell receptor and the phosphatidylinositol 3-kinase (PI3K) signalling pathways, together with their downstream partners, represent important therapeutic targets for B-cell lymphomas. Here, we evaluated the activity of acalabrutinib (ACP-196) and ACP-319 (AMG-319), second generation inhibitors of Bruton tyrosine kinase (BTK) and PI3Kδ inhibitor, respectively, in lymphoma pre-clinical models. The two compounds showed activity in activated B-cell-like diffuse large B-cell lymphoma (ABC DLBCL), mantle cell lymphoma and marginal zone lymphoma. Two in vivo experiments with ABC DLBCL and MCL xenografts confirmed the effect of the single agents. Benefit was achieved by exposing the lymphoma cell lines to both acalabrutinib and ACP-319. Two cell lines presented a discordant response to first and second generation BTK inhibitors, probably due to the inhibition by ibrutinib of kinases other than BTK. In conclusion, our data sustain the on-going current trials with acalabrutinib and ACP-319 as single agents and provide the basis for the investigation of their combination as well.

Biological Functions of Interferon ß-1a Are Enhanced By Deamidation.

Human type I Interferons (IFN-ß, IFN-ɛ, IFN-κ, IFN-ω, and 12 subtypes of IFN-α) are a family of pleiotropic cytokines with antiviral, antiproliferative, and immunomodulatory activities. They signal through the same cell surface receptors, IFNAR1 and IFNAR2, yet evoking markedly differential potency. One differentiating factor of IFN-ß from other type I interferons is the presence of a consensus sequence (NG) for deamidation. Comparing almost completely deamidated IFN-ß-1a with untreated IFN-ß-1a, this present study reports the increased activities in 3 in-vitro bioassays testing the antiviral, antiproliferative, and immunomodulatory properties, respectively, of the molecule. Deamidated IFN-ß-1a has the potential to improve current therapies in multiple sclerosis, and its ability to potentiate the MHC-Class I expression suggests a clinical benefit in diseases where the downmodulation of the MHC-class I expression plays a role (eg, in immuno-oncology combination therapies or antiviral agents). The present study on IFN-ß deamidation adds a new prospective on deamidation as part of a posttranslational modification code that allows the modulation of the biological properties of proteins. Moreover, it underlines the unique IFN-ß-1a properties that differentiate this molecule from other members of the type I interferon family.

Electrochemical treatment of water containing Microcystis aeruginosa in a fixed bed reactor with three-dimensional conductive diamond anodes.

An electrochemical treatment was investigated to remove Microcystis aeruginosa from water. A fixed bed reactor in flow was tested, which was equipped with electrodes constituted by stacks of grids electrically connected in parallel, with the electric field parallel to the fluid flow. Conductive diamond were used as anodes, platinised Ti as cathode. Electrolyses were performed in continuous and in batch recirculated mode with flow rates corresponding to Re from 10 to 160, current densities in the range 10-60Am(-2) and Cl(-) concentrations up to 600gm(-3). The absorbance of chlorophyll-a pigment and the concentration of products and by-products of electrolysis were measured. In continuous experiments without algae in the inlet stream, total oxidants concentrations as equivalent Cl2, of about 0.7gCl2m(-3) were measured; the maximum values were obtained at Re=10 and i=25Am(-2), with values strongly dependent on the concentration of Cl(-). The highest algae inactivation was obtained under the operative conditions of maximum generation of oxidants; in the presence of microalgae the oxidants concentrations were generally below the detection limit. Results indicated that most of the bulk oxidants electrogenerated is constituted by active chlorine. The prevailing mechanism of M. aeruginosa inactivation is the disinfection by bulk oxidants. The experimental data were quantitatively interpreted through a simple plug flow model, in which the axial dispersion accounts for the non-ideal flow behaviour of the system; the model was successfully used to simulate the performances of the reactor in the single-stack configuration used for the experiments and in multi-stack configurations.

In vitro biological characterization of IFN-ß-1a major glycoforms.

Recombinant human interferon ß-1a (IFN-ß-1a) is extensively used as the first-line treatment of relapsing forms of multiple sclerosis. Its glycosylation is recognized as having a complex impact on a wide range of molecule characteristics and functions. The present study reports the enrichment of IFN-ß-1a glycoforms and their physicochemical and biological characterization by means of electrospray ionization-mass spectrometry, sialic acid content, thermal denaturation and various in vitro bioassays (antiproliferative, antiviral, immunomodulatory and reporter gene assay). The glycoforms were fractionated by means of cation-exchange chromatography using recombinant IFN-ß-1a derived from Chinese Hamster Ovary cell culture as starting material. The obtained fractions contained bi- and higher-antennarity glycans as described in the European Pharmacopoeia monograph (Nr. 1639E, Interferon beta 1a concentrated solution). The in vitro bioassay responses revealed a correlation mainly with the glycan antennarity. It is therefore suggested that all glycoforms have biological activity and play a role in modulating the overall IFN-ß biological activity with higher-antennarity glycoforms being able to better sustain IFN-ß-1a bioactivity over time. These data indicate the role of IFN-ß-1a glycosylation in vivo and shed new light on the role of the glycosylation heterogeneity, in particular with regard to antennarity, on biological properties of glycoproteins.

Controlling the Er content of porous silicon using the doping current intensity.

The results of an investigation on the Er doping of porous silicon are presented. Electrochemical impedance spectroscopy, optical reflectivity, and spatially resolved energy dispersive spectroscopy (EDS) coupled to scanning electron microscopy measurements were used to investigate on the transient during the first stages of constant current Er doping. Depending on the applied current intensity, the voltage transient displays two very different behaviors, signature of two different chemical processes. The measurements show that, for equal transferred charge and identical porous silicon (PSi) layers, the applied current intensity also influences the final Er content. An interpretative model is proposed in order to describe the two distinct chemical processes. The results can be useful for a better control over the doping process. PACS: 81.05.Rm; 82.45.Rr.

Electrochemical treatment of phenolic waters in presence of chloride with boron-doped diamond (BDD) anodes: experimental study and mathematical model.

This work deals with an experimental and numerical study on the electrochemical treatment of waters containing phenolic compounds with boron-doped diamond (BDD) anodes. Anodic oxidation of m-cresol, as a model of phenolic compound, was investigated by galvanostatic electrolyses. The electrolyses were carried out under different experimental conditions by using an impinging-jet flow cell inserted in a hydraulic circuit in a closed loop. On the basis of the experimental results a mathematical model was implemented to simulate the effect of the chemistry of organic compounds and solution on the process, in particular the effect of chlorides on the kinetics of m-cresol oxidation. The effect of hydrodynamics of the cell on the mass transfer towards the electrode surface was also considered. The model was validated through comparison with experimental data: the results showed that the proposed model well interpreted the complex effect on removal efficiency of such operative parameters as current density, hydrodynamic of the reactor and chemistry of the solution. The model predictions were utilised to obtain quantitative information on the reaction mechanism, as well as to predict the performance of the process under different operative conditions, by calculating some relevant figures of merit.

Electrokinetic removal of 2,6-dichlorophenol and diuron from kaolinite and humic acid-clay system.

This paper presents the results of a study on the electrokinetic treatment of kaolinite and humic acid kaolinite complexes spiked with 2,6-dichlorophenol or 3-(3,4-dichlorophenyl)-1,1-dimethylurea (diuron). In particular, the attention was paid to the interaction between solid surface and dissolved organics: the effects of contaminant sorption as well as the physicochemical reactions on the efficiency of electrokinetic remediation were investigated. Using a 3V/cm voltage gradient, approximately 90% of diuron was removed from kaolinite after one water pore volume was collected in the cathode reservoir, but much lower efficiency was obtained in the electrokinetic removal of this compound from humic acid-coated kaolinite. The results also showed that partial degradation of the contaminant occurred during electrokinetic treatment of kaolinite clay spiked with 2,6-dichlorophenol: the contamination in the clay could be remediated by the combination of electrokinetic extraction and electrochemical reactions.

Electrochemical oxidation of phenolic and other organic compounds at boron doped diamond electrodes for wastewater treatment: effect of mass transfer.

The paper presents the results of an experimental study on oxidation at boron doped diamond electrodes (BDD) of some phenolic compounds: phenol (PH), para-hydroxibenzoic acid (PHB), cathecole (CT), hydroquinone (HQ) are considered, singularly contained in aqueous solutions or in the presence of glucose (G), which was selected to represent the class of biodegradable compounds. Oxidation of benzoquinone (BQ) and maleic acid (MA), generally detected as intermediates products from phenol degradation, is also investigated. Great attention is paid to verify the feasibility of a selective process in which the oxidation is specifically addressed to the phenolic fraction up to non toxic intermediate products which are more biodegradable than the original phenols.

Electrochemical oxidation of p-hydroxybenzoic and protocathecuic acids at a dimensional stable anode (DSA) in the presence of NaCl.

This work is part of a wider research programme on innovative technologies for industrial wastewater treatment. Results from electrolyses at DSA commercial anodes of synthetic solutions with composition analogous to that of agro-industrial wastes are presented. The results obtained indicate that the rate of degradation of phenolic compounds is high, provided that chloride ions are present in solution. Oxidation of phenolic compounds is faster than that of biodegradable substances, such as sugars or amminoacids. Moreover, investigation on the trend of toxicity during the treatment, seems to exclude that toxic intermediates persist in solution when phenolic compounds are removed. Experiments on olive oil mill wastewater (OMW) samples show that the results obtained from synthetic solutions are extensible to real wastewater. When phenolic compounds are completely removed, the toxicity of the solution is very low; the initial dark colour of the solution, due to the brown pigment which characterises OMW, is nearly completely disappeared.

A comparison of the biologic activity of two recombinant IFN-beta preparations used in the treatment of relapsing-remitting multiple sclerosis.

Recent clinical trials with interferon-beta (IFN-beta) in relapsing-remitting multiple sclerosis (RRMS) have clearly demonstrated that the IFN-beta dosing regimen affects the clinical efficacy, thereby highlighting the importance of determining the relative biologic activities of the IFN-beta products currently available. Although studies have been published that examine the biologic activities of the two structurally different forms of recombinant IFN-beta, IFN-beta1a (Rebif), Serono, Geneva, Switzerland) and IFN-beta1b (Betaseron)/Betaferon), Berlex [Montville, NJ]/Schering [Berlin, Germany]), there have been few direct comparative studies. Therefore, to obtain a more accurate estimate of the relative biologic activities of Rebif and Betaseron, this study examined the antiviral activities of these two products within the same assay system and against the same natural human IFN-beta standard. Whereas the manufacturers' information suggests that the bioactivity of Betaseron is only about 8.7-fold less than that of Rebif, the results of the present direct, comparative study show that Rebif has an antiviral activity 14 times greater than that of Betaseron. This may have important clinical implications, because on the basis of the results reported here, Rebif at 44 microg t.i.w. is approximately double the maximal licensed weekly dose for Betaseron.