Publications by authors named "Daniela Berger"

34 Publications

Culturing cells with mast cell phenotype and function: Comparison of peripheral blood and bone marrow as a source.

J Immunol Methods 2021 Apr 30;495:113061. Epub 2021 Apr 30.

University of Antwerp, Faculty of Medicine and Health Sciences, Department of Immunology, Allergology, Rheumatology and the Infla-Med Centre of Excellence, Antwerp (Belgium) and Immunology, Allergology, Rheumatology, Antwerp University Hospital, Antwerp, Belgium; Department of Immunology and Allergology, AZ Jan Palfijn Gent, Ghent, Belgium.

Background: Studies on the mechanisms that govern mast cell (MC) functions are hindered by the difficulties in isolating sufficient numbers of these tissue-resident cells. Therefore, many research groups use cultured human MCs obtained out of progenitor cells. However, these culture methods significantly differ regarding primary source material, culture durations and conditions. Consequently, the finally obtained cells are likely to exhibit morphological, phenotypical and/or functional heterogeneity.

Objective: To compare the phenotype and functionality of cells cultured from peripheral blood and bone marrow progenitor cells from patients with suspected clonal MC disease. These cells are designated as PBCMCs and BMCMCs, respectively.

Methods: Twenty paired PBCMCs and BMCMCs cultures starting from CD34 progenitor cells were compared. Cells were cultured for 4 weeks. Phenotyping included Giemsa and CD117 staining and flow cytometric staining for CD117, CD203c, FcεRI, MRGPRX2, CD300a, CD32, CD63 and CD25. Functional assessment included measurement of the up-regulation of CD63 after cross-linking of the high affinity receptor for IgE (FcεRI) with anti-FcεRI and ligation of MRGPRX2 with substance P.

Results: PBCMCs and BMCMCs are phenotypically comparable. Functionally, after activation with anti-FcεRI and substance P, PBCMCs and BMCMCs show similar up-regulation of the lysosomal degranulation marker CD63. However, the yield of PBCMCs is higher than BMCMs and peripheral blood cultures are purer than bone marrow cultures.

Conclusion: PBCMCs are an attractive alternative to the more difficult to obtain BMCMCs for the exploration of the complex mechanisms that govern IgE- and MRGPRX2-dependent MC activation and degranulation. Unlike BMCMCs, PBCMCs are easily accessible and enable repetitive analyses.
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http://dx.doi.org/10.1016/j.jim.2021.113061DOI Listing
April 2021

Phenotypic characterization of leukemia-initiating stem cells in chronic myelomonocytic leukemia.

Leukemia 2021 Mar 30. Epub 2021 Mar 30.

Department of Medicine I, Division of Hematology and Hemostaseology, Medical University of Vienna, Vienna, Austria.

Chronic myelomonocytic leukemia (CMML) is a stem cell-derived neoplasm characterized by dysplasia, uncontrolled expansion of monocytes, and substantial risk to transform to secondary acute myeloid leukemia (sAML). So far, little is known about CMML-initiating cells. We found that leukemic stem cells (LSC) in CMML reside in a CD34/CD38 fraction of the malignant clone. Whereas CD34/CD38 cells engrafted NSGS mice with overt CMML, no CMML was produced by CD34/CD38 progenitors or the bulk of CD34 monocytes. CMML LSC invariably expressed CD33, CD117, CD123 and CD133. In a subset of patients, CMML LSC also displayed CD52, IL-1RAP and/or CLL-1. CMML LSC did not express CD25 or CD26. However, in sAML following CMML, the LSC also expressed CD25 and high levels of CD114, CD123 and IL-1RAP. No correlations between LSC phenotypes, CMML-variant, mutation-profiles, or clinical course were identified. Pre-incubation of CMML LSC with gemtuzumab-ozogamicin or venetoclax resulted in decreased growth and impaired engraftment in NSGS mice. Together, CMML LSC are CD34/CD38 cells that express a distinct profile of surface markers and target-antigens. During progression to sAML, LSC acquire or upregulate certain cytokine receptors, including CD25, CD114 and CD123. Characterization of CMML LSC should facilitate their enrichment and the development of LSC-eradicating therapies.
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http://dx.doi.org/10.1038/s41375-021-01227-zDOI Listing
March 2021

Extracellular matrix biomimetic polymeric membranes enriched with silver nanoparticles for wound healing.

Biomed Mater 2021 Feb 11. Epub 2021 Feb 11.

Cellular and Molecular Biology, National Institute of Research and Development for Biological Sciences, 296, Sp Indepedentei, Bucharest, Bucharest, 060031, ROMANIA.

Severe skin injuries, including burns, represent a real concern for the global health-care system and therefore, there is an increased interest in developing wound dressings, in order to stimulate and enhance skin tissue repair. The aim of this study was to design novel hybrid materials, biomimetic to skin extracellular matrix and enriched with silver nanoparticles (nAg), in order to provide both dermal tissue regeneration and antimicrobial activity. Two material variants (variant A and variant B) consisting of type I collagen (COL), chondroitin sulfate (CS) and k-elastin peptides (EL) enriched with positively-charged nAg, were conditioned as membranes. UV exposure ensured both sterilisation and cross-linking of the materials. Physico-chemical characterization of the hybrid biomaterials showed values of density and swelling degree higher than those of COL membrane, while the process of in vitro degradation followed a similar pattern. Infrared spectroscopy and X-ray diffraction indicated alterations of the characteristic structural features and crystallinity of COL after blending with CS and EL and nAg embedding. Scanning electron microscopy observations revealed different surface morphologies of the hybrid membranes, according to their composition. In vitro studies on L929 fibroblasts and HaCaT keratinocytes showed that both hybrid membranes exhibited good cytocompatibility and promoted higher cell proliferation compared to COL sample, as evaluated by MTT and Live/Dead assays. The presence of actin filaments highlighted by fluorescent labelling confirmed the fibroblast and keratinocyte adhesion onto the surface of hybrid membranes. Most importantly, both materials showed an increased wound healing ability in an in vitro scratch assay model, stimulating cell migration at 24 h post-seeding. In addition, good antimicrobial activity was recorded, especially against Gram-positive bacterial strain. Altogether, our findings recommend COL-CS-EL-nAg hybrid membranes as good candidates for wound healing acceleration and bioengineering of skin tissue.
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http://dx.doi.org/10.1088/1748-605X/abe55dDOI Listing
February 2021

A Review of Composite Phase Change Materials Based on Porous Silica Nanomaterials for Latent Heat Storage Applications.

Molecules 2021 Jan 5;26(1). Epub 2021 Jan 5.

Faculty of Applied Chemistry and Material Science, University "Politehnica" of Bucharest, 1-7 Polizu Street, 011061 Bucharest, Romania.

Phase change materials (PCMs) can store thermal energy as latent heat through phase transitions. PCMs using the solid-liquid phase transition offer high 100-300 J g enthalpy at constant temperature. However, pure compounds suffer from leakage, incongruent melting and crystallization, phase separation, and supercooling, which limit their heat storage capacity and reliability during multiple heating-cooling cycles. An appropriate approach to mitigating these drawbacks is the construction of composites as shape-stabilized phase change materials which retain their macroscopic solid shape even at temperatures above the melting point of the active heat storage compound. Shape-stabilized materials can be obtained by PCMs impregnation into porous matrices. Porous silica nanomaterials are promising matrices due to their high porosity and adsorption capacity, chemical and thermal stability and possibility of changing their structure through chemical synthesis. This review offers a first in-depth look at the various methods for obtaining composite PCMs using porous silica nanomaterials, their properties, and applications. The synthesis and properties of porous silica composites are presented based on the main classes of compounds which can act as heat storage materials (paraffins, fatty acids, polymers, small organic molecules, hydrated salts, molten salts and metals). The physico-chemical phenomena arising from the nanoconfinement of phase change materials into the silica pores are discussed from both theoretical and practical standpoints. The lessons learned so far in designing efficient composite PCMs using porous silica matrices are presented, as well as the future perspectives on improving the heat storage materials.
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http://dx.doi.org/10.3390/molecules26010241DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7796474PMC
January 2021

Biological Evaluation of Black Chokeberry Extract Free and Embedded in Two Mesoporous Silica-Type Matrices.

Pharmaceutics 2020 Sep 1;12(9). Epub 2020 Sep 1.

Department of Pharmacognosy, "Victor Babeş" University of Medicine and Pharmacy, Eftimie Murgu Square, No. 2, 300041 Timisoara, Romania.

Black chokeberry fruits possess a wide range of biological activities, among which the most important that are frequently mentioned in the literature are their antioxidant, anti-inflammatory, anti-proliferative, and antimicrobial properties. The present paper reports, for the first time, the encapsulation of the ethanolic extract of L. fruits into two mesoporous silica-type matrices (i.e., pristine MCM-41 and MCM-41 silica decorated with zinc oxide nanoparticles). The aim of this work was to evaluate the antiradicalic capacity, the antimicrobial potential, and the effects on the cell viability on a cancer cell line (i.e., A375 human melanoma cell line) versus normal cells (i.e., HaCaT human keratinocytes) of black chokeberry extract loaded on silica-type matrices in comparison to that of the extract alone. The ethanolic polyphenolic extract obtained by conventional extraction was characterized by high-performance liquid chromatography with a photodiode array detector (HPLC-PDA) and spectrophotometric methods. The extract was found to contain high amounts of polyphenols and flavonoids, as well as good radical scavenging activity. The extract-loaded materials were investigated by Fourier transform infrared spectroscopy, N adsorption-desorption isotherms, thermal analysis, and radical scavenger activity on solid samples. The black chokeberry extract, both free and loaded onto mesoporous silica-type matrices, exhibited a significant antioxidant capacity. Antibacterial activity was recorded only for Gram-positive bacteria, with a more potent antibacterial effect being observed for the extract loaded onto the ZnO-modified MCM-41 silica-type support than for the free extract, probably due to the synergistic effect of the ZnO nanoparticles that decorate the pore walls of silica. The cellular viability test (i.e., MTT assay) showed dose- and time-dependent activity regarding the melanoma cell line. The healthy cells were less affected than the cancer cells, with all tested samples showing good cytocompatibility at doses of up to 100 µg/mL. Improved in vitro antiproliferative and antimigratory (i.e., scratch assay) potential was demonstrated through the loading of black chokeberry extract into mesoporous silica-type matrices, and the screened samples exhibited low selectivity against the tested non-tumor cell line. Based on presented results, one can conclude that mesoporous silica-type matrices are good hosts for black chokeberry extract, increasing its antioxidant, antibacterial (on the screened strains), and in vitro antitumor (on the screened cell line) properties.
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http://dx.doi.org/10.3390/pharmaceutics12090838DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7558869PMC
September 2020

Effect of Nanoconfinement of Polyphenolic Extract from Grape Pomace into Functionalized Mesoporous Silica on Its Biocompatibility and Radical Scavenging Activity.

Antioxidants (Basel) 2020 Aug 3;9(8). Epub 2020 Aug 3.

Faculty of Pharmacy, "Ovidius" University of Constanta, Aleea Universitatii No. 1, 900470 Constanta, Romania.

The aim of this paper is to assess the properties of Mamaia (MM) grape pomace polyphenolic extract loaded onto pristine and functionalized MCM-41 mesoporous silica as potential ingredients for nutraceuticals or cosmetics. The chemical profile of hydroalcoholic polyphenolic extracts, prepared either by conventional extraction or microwave-assisted method, was analyzed by reverse-phase high-performance liquid chromatography with photodiode array detector (HPLC-PDA) analysis, while their radical scavenger activity (RSA) was evaluated using DPPH (2,2-diphenyl-1-picrylhydrazyl radical) and ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) assays. The extract-loaded materials were characterized by Fourier transform infrared (FTIR) spectroscopy, N adsorption-desorption isotherms, thermogravimetric analysis, as well as RSA (DPPH and ABTS assays). The polyphenols release profiles from pristine and functionalized (with mercaptopropyl, propyl sulfonic acid, cyanoethyl and propionic acid moieties) MCM-41-type supports were determined in phosphate buffer solution (PBS) pH 5.7. For selected materials containing embedded phytochemicals, cellular viability, and oxidative stress level on immortalized mouse embryonic fibroblast cell line (NIH3T3) were evaluated. A more acidic functional groups linked on silica pore walls determined a higher amount of phytochemicals released in PBS. The extract-loaded materials showed a good cytocompatibility on tested concentrations. The embedded extract preserved better the RSA over time than the free extract. The polyphenols-loaded MCM-41-type silica materials, especially [email protected] material, demonstrated a good in vitro antioxidant effect on NIH3T3 cells, being potential candidates for nutraceutical or cosmetic formulations.
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http://dx.doi.org/10.3390/antiox9080696DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7465047PMC
August 2020

Properties of L. and L. Extracts Free and Embedded into Mesopores of Silica and Titania Nanomaterials.

Nanomaterials (Basel) 2020 Apr 25;10(5). Epub 2020 Apr 25.

Department of Pharmacognosy, University of Medicine and Pharmacy "Victor Babes", Eftimie Murgu Square No. 2, 300041 Timisoara, Romania.

This study evidenced the nanoconfinement effect on polyphenolic extracts prepared from L. and L. into the mesopores of silica and titania nanomaterials on their radical scavenging capacity and antimicrobial potential. The ethanolic and hydroalcoholic extracts obtained either by conventional or microwave-assisted extraction were characterized in terms of total polyphenols, total flavonoids, and chlorophyll content, as well as radical scavenging activity by consecrated spectrometric determinations. The phytochemical fingerprint of extracts was analyzed by high-performance liquid chromatography-photodiode array detector. extracts exhibited better radical scavenging capacity and antimicrobial potential than extracts. The mesoporous MCM-41 silica and titania nanomaterials, prepared by the sol-gel method, were characterized by small- and wide-angle powder diffraction, FTIR spectroscopy, nitrogen adsorption-desorption isotherms, scanning electron microscopy and transmission electron microscopy, while the materials containing embedded extracts were analyzed through Fourier-transform infrared spectroscopy, N sorption measurements, and thermal analysis. All extracts free and embedded in mesoporous matrix exhibited high radical scavenger properties and good bactericidal activity against several reference strains. It was proved that by embedding the polyphenolic extracts into mesopores of silica or titania nanoparticles, the phytochemicals stability was enhanced as the materials containing extract exhibited higher radical scavenger activity after 3-6 months storage than that of the free extracts. Additionally, the extract-loaded material showed mild improved antimicrobial activity in comparison with the corresponding free extract.
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http://dx.doi.org/10.3390/nano10050820DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7712395PMC
April 2020

STAT5 is Expressed in CD34/CD38 Stem Cells and Serves as a Potential Molecular Target in Ph-Negative Myeloproliferative Neoplasms.

Cancers (Basel) 2020 Apr 21;12(4). Epub 2020 Apr 21.

Ludwig Boltzmann Institute for Hematology and Oncology, Medical University of Vienna, 1090 Vienna, Austria.

Janus kinase 2 (JAK2) and signal transducer and activator of transcription-5 (STAT5) play a key role in the pathogenesis of myeloproliferative neoplasms (MPN). In most patients, V617F or mutations are found and lead to activation of various downstream signaling cascades and molecules, including STAT5. We examined the presence and distribution of phosphorylated (p) STAT5 in neoplastic cells in patients with MPN, including polycythemia vera (PV, = 10), essential thrombocythemia (ET, = 15) and primary myelofibrosis (PMF, = 9), and in the V617F-positive cell lines HEL and SET-2. As assessed by immunohistochemistry, MPN cells displayed pSTAT5 in all patients examined. Phosphorylated STAT5 was also detected in putative CD34/CD38 MPN stem cells (MPN-SC) by flow cytometry. Immunostaining experiments and Western blotting demonstrated pSTAT5 expression in both the cytoplasmic and nuclear compartment of MPN cells. Confirming previous studies, we also found that JAK2-targeting drugs counteract the expression of pSTAT5 and growth in HEL and SET-2 cells. Growth-inhibition of MPN cells was also induced by the STAT5-targeting drugs piceatannol, pimozide, AC-3-019 and AC-4-130. Together, we show that CD34/CD38 MPN-SC express pSTAT5 and that pSTAT5 is expressed in the nuclear and cytoplasmic compartment of MPN cells. Whether direct targeting of pSTAT5 in MPN-SC is efficacious in MPN patients remains unknown.
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http://dx.doi.org/10.3390/cancers12041021DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7225958PMC
April 2020

Mesoporous Cobalt Ferrite Nanosystems Obtained by Surfactant-Assisted Hydrothermal Method: Tuning Morpho-structural and Magnetic Properties via pH-Variation.

Nanomaterials (Basel) 2020 Mar 6;10(3). Epub 2020 Mar 6.

National Institute of Materials Physics, Atomistilor 405A, 077125 Magurele, Romania.

A facile and cheap surfactant-assisted hydrothermal method was used to prepare mesoporous cobalt ferrite nanosystems with BET surface area up to 151 m/g. These mesostructures with high BET surface areas and pore sizes are made from assemblies of nanoparticles (NPs) with average sizes between 7.8 and 9.6 nm depending on the initial pH conditions. The pH proved to be the key factor for controlling not only NP size, but also the phase purity and the porosity properties of the mesostructures. At pH values lower than 7, a parasite hematite phase begins to form. The sample obtained at pH = 7.3 has magnetization at saturation M = 38 emu/g at 300 K (54.3 emu/g at 10 K) and BET surface area S = 151 m/g, whereas the one obtained at pH = 8.3 has M = 68 emu/g at 300 K (83.6 emu/g at 10 K) and S = 101 m/g. The magnetic coercive field values at 10 K are high at up to 12,780 Oe, with a maximum coercive field reached for the sample obtained at pH = 8.3. Decreased magnetic performances are obtained at pH values higher than 9. The iron occupancies of the tetrahedral and octahedral sites belonging to the cobalt ferrite spinel structure were extracted through decomposition of the Mössbauer patterns in spectral components. The magnetic anisotropy constants of the investigated NPs were estimated from the temperature dependence of the hyperfine magnetic field. Taking into consideration the high values of BET surface area and the magnetic anisotropy constants as well as the significant magnetizations for saturation at ambient temperature, and the fact that all parameters can be adjusted through the initial pH conditions, these materials are very promising as recyclable anti-polluting agents, magnetically separable catalysts, and targeted drug delivery vehicles.
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http://dx.doi.org/10.3390/nano10030476DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7153623PMC
March 2020

Polyphenolic Extract from L. Leaves Free and Loaded into Lipid Vesicles.

Nanomaterials (Basel) 2019 Dec 25;10(1). Epub 2019 Dec 25.

Faculty of Applied Chemistry and Materials Science, University "Politehnica" of Bucharest, 1-7 Gheorghe Polizu St., 011061 Bucharest, Romania.

The paper deals with the preparation and characterisation of hydroalcoholic polyphenolic extract from (SE) leaves that was further loaded into three-types of lipid vesicles: liposomes, transfersomes, and ethosomes, to improve its bioavailability and achieve an optimum pharmacological effect. For L.-loaded lipid vesicles, the entrapment efficiency, particle size, polydispersity index and stability were determined. All prepared lipid vesicles showed a good entrapment efficiency, in the range of 75-85%, nanometric size, low polydispersity indexes, and good stability over three months at 4 °C. The in vitro polyphenols released from lipid vehicles demonstrated slower kinetics when compared to the free extract dissolution in phosphate buffer solution at pH 7.4. Either free SE extract or SE extract loaded into lipid vesicles demonstrated a cytoprotective effect, even at low concentration, 5 ug/mL, against hydrogen peroxide-induced toxicity on L-929 mouse fibroblasts' cell lines. However, the cytoprotective effect depended on the time of the cells pre-treatment with SE extract before exposure to a hydrogen peroxide solution of 50 mM concentration, requiring at least 12 h of pre-treatment with polyphenols with radical scavenging capacity.
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http://dx.doi.org/10.3390/nano10010056DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7023427PMC
December 2019

CDK4/CDK6 inhibition as a novel strategy to suppress the growth and survival of BCR-ABL1+ clones in TKI-resistant CML.

EBioMedicine 2019 Dec 21;50:111-121. Epub 2019 Nov 21.

Department of Internal Medicine I, Division of Hematology & Hemostaseology, Medical University of Vienna, Waehringer Guertel 18-20, 1090 Vienna, Austria; Ludwig Boltzmann Institute for Hematology & Oncology, Medical University of Vienna, Austria. Electronic address:

Purpose: Ponatinib is the only approved tyrosine kinase inhibitor (TKI) suppressing BCR-ABL1-mutated cells in chronic myeloid leukemia (CML). However, due to side effects and resistance, BCR-ABL1-mutated CML remains a clinical challenge. Hydroxyurea (HU) has been used for cytoreduction in CML for decades. We found that HU suppresses or even eliminates BCR-ABL1+ sub-clones in heavily pretreated CML patients. Based on this observation, we investigated the effects of HU on TKI-resistant CML cells in vitro.

Methods: Viability, apoptosis and proliferation of drug-exposed primary CML cells and BCR-ABL1+ cell lines were examined by flow cytometry and H-thymidine-uptake. Expression of drug targets was analyzed by qPCR and Western blotting.

Findings: HU was more effective in inhibiting the proliferation of leukemic cells harboring BCR-ABL1 or T315I-including compound-mutations compared to cells expressing wildtype BCR-ABL1. Moreover, HU synergized with ponatinib and ABL001 in inducing growth inhibition in CML cells. Furthermore, HU blocked cell cycle progression in leukemic cells, which was accompanied by decreased expression of CDK4 and CDK6. Palbociclib, a more specific CDK4/CDK6-inhibitor, was also found to suppress proliferation in primary CML cells and to synergize with ponatinib in producing growth inhibition in BCR-ABL1+ cells, suggesting that suppression of CDK4/CDK6 may be a promising concept to overcome BCR-ABL1-associated TKI resistance.

Interpretation: HU and the CDK4/CDK6-blocker palbociclib inhibit growth of CML clones expressing BCR-ABL1 or complex T315I-including compound-mutations. Clinical studies are required to confirm single drug effects and the efficacy of `ponatinib+HU´ and ´ponatinib+palbociclib´ combinations in advanced CML.

Funding: This project was supported by the Austrian Science Funds (FWF) projects F4701-B20, F4704-B20 and P30625.
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http://dx.doi.org/10.1016/j.ebiom.2019.11.004DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6921367PMC
December 2019

Physicochemical and Biological Properties of Gelatin Extracted from Marine Snail .

Mar Drugs 2019 Oct 17;17(10). Epub 2019 Oct 17.

Departament of Cellular and Molecular Biology, National Institute of R&D for Biological Sciences, 296 Splaiul Independentei, 060031 Bucharest, Romania.

In this study, we aimed to obtain gelatin from the marine snail using acidic and enzymatic extraction methods and to characterize these natural products for cosmetic and pharmaceutical applications. Marine gelatins presented protein values and hydroxyproline content similar to those of commercial mammalian gelatin, but with higher melting temperatures. Their electrophoretic profile and Fourier transform infrared (FTIR) spectra revealed protein and absorption bands situated in the amide region, specific for gelatin molecule. Scanning electron microscopy (SEM) analysis showed significant differences in the structure of the lyophilized samples, depending on the type of gelatin. In vitro studies performed on human keratinocytes showed no cytotoxic effect of acid-extracted gelatin at all tested concentrations and moderate cytotoxicity of enzymatic extracted gelatin at concentrations higher than 0.5 mg/mL. Also, both marine gelatins favored keratinocyte cell adhesion. No irritant potential was recorded as the level of IL-1α and IL-6 proinflammatory cytokines released by HaCaT cells cultivated in the presence of marine gelatins was significantly reduced. Together, these data suggest that marine snails are an alternative source of gelatins with potential use in pharmaceutical and skincare products.
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http://dx.doi.org/10.3390/md17100589DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6835507PMC
October 2019

Polyphenols extract from grape pomace. Characterization and valorisation through encapsulation into mesoporous silica-type matrices.

Food Chem Toxicol 2019 Nov 23;133:110787. Epub 2019 Aug 23.

University "Politehnica" of Bucharest, Faculty of Applied Chemistry and Material Sciences, 1-7 Gheorghe Polizu Street, Bucharest, 011061, Romania. Electronic address:

We report the encapsulation of two grape pomace polyphenolic extracts into mesoporous MCM-41-type silica matrices (pristine and Zn or Mg heteroatom modified) to reduce the extract sensitivity and enhance its stability, while preserving the radical scavenger activity. Various grapes marc (Cabernet Saugvinon and Feteasca Neagra from the Black Sea region and commercially available grape skins powder) were used to prepare ethanolic extracts either through conventional extraction, or microwave-assisted procedure. The polyphenolic extracts composition was analysed by reversed phase-high pressure liquid chromatography and spectrometric determination of total polyphenols and ascorbic acid (using Folin Ciocalteu reagent), total flavonoids (by AlCl complexation), as well as total anthocyanin monomeric pigments content. The encapsulated extract into MCM-41 silica, as well as Zn-MCM-41 and Mg-MCM-41 matrices showed an enhanced radical scavenger activity assessed by DPPH procedure developed for solid samples. The cytocompatibility tests performed on HaCaT keratinocyte human cells demonstrated a good cytocompatibility for the Cabernet Saugvinon and grape skins extracts free and encapsulated into MCM-41-type matrices.
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http://dx.doi.org/10.1016/j.fct.2019.110787DOI Listing
November 2019

Identification of a leukemia-initiating stem cell in human mast cell leukemia.

Leukemia 2019 11 5;33(11):2673-2684. Epub 2019 Apr 5.

Ludwig Boltzmann Institute for Hematology and Oncology, Medical University of Vienna, 1090, Vienna, Austria.

Mast cell leukemia (MCL) is a highly fatal malignancy characterized by devastating expansion of immature mast cells in various organs. Although considered a stem cell disease, little is known about MCL-propagating neoplastic stem cells. We here describe that leukemic stem cells (LSCs) in MCL reside within a CD34/CD38 fraction of the clone. Whereas highly purified CD34/CD38 cells engrafted NSG mice with fully manifesting MCL, no MCL was produced by CD34/CD38 progenitors or the bulk of KIT/CD34 mast cells. CD34/CD38 MCL cells invariably expressed CD13 and CD133, and often also IL-1RAP, but did not express CD25, CD26 or CLL-1. CD34/CD38 MCL cells also displayed several surface targets, including CD33, which was homogenously expressed on MCL LSCs in all cases, and the D816V mutant form of KIT. Although CD34/CD38 cells were resistant against single drugs, exposure to combinations of CD33-targeting and KIT-targeting drugs resulted in LSC-depletion and markedly reduced engraftment in NSG mice. Together, MCL LSCs are CD34/CD38 cells that express distinct profiles of markers and target antigens. Characterization of MCL LSCs should facilitate their purification and should support the development of LSC-eradicating curative treatment approaches in this fatal type of leukemia.
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http://dx.doi.org/10.1038/s41375-019-0460-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6839966PMC
November 2019

A kinase profile-adapted drug combination elicits synergistic cooperative effects on leukemic cells carrying BCR-ABL1 in Ph+ CML.

Leuk Res 2019 03 28;78:36-44. Epub 2018 Dec 28.

Department of Internal Medicine I, Division of Hematology & Hemostaseology, Medical University of Vienna, Austria; Ludwig Boltzmann Institute for Hematology and Oncology, Medical University of Vienna, Austria. Electronic address:

In chronic myeloid leukemia (CML), resistance against second-generation tyrosine kinase inhibitors (TKI) remains a serious clinical challenge, especially in the context of multi-resistant BCR-ABL1 mutants, such as T315I. Treatment with ponatinib may suppress most of these mutants, including T315I, but is also associated with a high risk of clinically relevant side effects. We screened for alternative treatment options employing available tyrosine kinase inhibitors (TKI) in combination. Dasatinib and bosutinib are two second-generation TKI that bind to different, albeit partially overlapping, spectra of kinase targets in CML cells. This observation prompted us to explore anti-leukemic effects of the combination dasatinib + bosutinib in highly resistant primary CML cells, various CML cell lines (K562, K562R, KU812, KCL22) and Ba/F3 cells harboring various BCR-ABL1 mutant-forms. We found that bosutinib synergizes with dasatinib in inducing growth inhibition and apoptosis in all CML cell lines and in Ba/F3 cells exhibiting BCR-ABL1. Clear synergistic effects were also observed in primary CML cells in all patients tested (n = 20), including drug-resistant cells carrying BCR-ABL1. Moreover, the drug combination produced cooperative or even synergistic apoptosis-inducing effects on CD34/CD38 CML stem cells. Finally, we found that the drug combination is a potent approach to block the activity of major additional CML targets, including LYN, KIT and PDGFRα. Together, bosutinib and dasatinib synergize in producing anti-leukemic effects in drug-resistant CML cells. Whether such cooperative TKI effects also occur in vivo in patients with drug-resistant CML, remains to be determined in forthcoming studies.
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http://dx.doi.org/10.1016/j.leukres.2018.12.013DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6834439PMC
March 2019

Oral Biofilms: Development, Control, and Analysis.

High Throughput 2018 Aug 31;7(3). Epub 2018 Aug 31.

Department of Pharmaceutical and Biomedical Sciences, Touro College of Pharmacy, New York, NY 10027, USA.

The oral cavity harbors hundreds of microbial species that are present either as planktonic cells or incorporated into biofilms. The majority of the oral microbes are commensal organisms. Those that are pathogenic microbes can result in oral infections, and at times can initiate systemic diseases. Biofilms that contain pathogens are challenging to control. Many conventional antimicrobials have proven to be ineffective. Recent advances in science and technology are providing new approaches for pathogen control and containment and methods to characterize biofilms. This perspective provides (1) a general understanding of biofilm development; (2) a description of emerging chemical and biological methods to control oral biofilms; and (3) an overview of high-throughput analytical approaches to analyze biofilms.
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http://dx.doi.org/10.3390/ht7030024DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6163956PMC
August 2018

CD44 is a RAS/STAT5-regulated invasion receptor that triggers disease expansion in advanced mastocytosis.

Blood 2018 11 17;132(18):1936-1950. Epub 2018 Jul 17.

Division of Hematology and Hemostaseology, Department of Internal Medicine I, Medical University of Vienna, Vienna, Austria.

The Hermes receptor CD44 is a multifunctional adhesion molecule that plays an essential role in the homing and invasion of neoplastic stem cells in various myeloid malignancies. Although mast cells (MCs) reportedly express CD44, little is known about the regulation and function of this receptor in neoplastic cells in systemic mastocytosis (SM). We found that clonal CD34/CD38 stem cells, CD34/CD38 progenitor cells, and CD117/CD34 MCs invariably express CD44 in patients with indolent SM (ISM), SM with an associated hematologic neoplasm, aggressive SM, and MC leukemia (MCL). In addition, all human MCL-like cell lines examined (HMC-1, ROSA, and MCPV-1) displayed cytoplasmic and cell-surface CD44. We also found that expression of CD44 in neoplastic MCs depends on RAS-MEK and STAT5 signaling and increases with the aggressiveness of SM. Correspondingly, higher levels of soluble CD44 were measured in the sera of patients with advanced SM compared with ISM or cutaneous mastocytosis and were found to correlate with overall and progression-free survival. To investigate the functional role of CD44, a xenotransplantation model was employed using severe combined immunodeficient (SCID) mice, HMC-1.2 cells, and a short hairpin RNA (shRNA) against CD44. In this model, the shRNA-mediated knockdown of CD44 resulted in reduced MC expansion and tumor formation and prolonged survival in SCID mice compared with HMC-1.2 cells transduced with control shRNA. Together, our data show that CD44 is a RAS-MEK/STAT5-driven MC invasion receptor that correlates with the aggressiveness of SM. Whether CD44 can serve as therapeutic target in advanced SM remains to be determined in forthcoming studies.
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http://dx.doi.org/10.1182/blood-2018-02-833582DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6382065PMC
November 2018

Hitting two oncogenic machineries in cancer cells: cooperative effects of the multi-kinase inhibitor ponatinib and the BET bromodomain blockers JQ1 or dBET1 on human carcinoma cells.

Oncotarget 2018 May 29;9(41):26491-26506. Epub 2018 May 29.

Ludwig Boltzmann Cluster Oncology, Medical University of Vienna, Vienna, Austria.

In recent years, numerous new targeted drugs, including multi-kinase inhibitors and epigenetic modulators have been developed for cancer treatment. Ponatinib blocks a variety of tyrosine kinases including ABL and fibroblast growth factor receptor (FGFR), and the BET bromodomain (BRD) antagonists JQ1 and dBET1 impede MYC oncogene expression. Both drugs have demonstrated substantial anti-cancer efficacy against several hematological malignancies. Solid tumors, on the other hand, although frequently driven by FGFR and/or MYC, are often unresponsive to these drugs. This is due, at least in part, to compensatory feedback-loops in the kinome and transcription network of these tumors, which are activated in response to drug exposure. Therefore, we hypothesized that the combination of the multi-kinase inhibitor ponatinib with transcription modulators such as JQ1 or dBET1 might overcome this therapeutic recalcitrance. Using H-thymidine uptake, cell cycle analysis, and caspase-3 or Annexin V labeling, we demonstrate that single drugs induce moderate dose-dependent growth-inhibition and/or apoptosis in colon (HCT116, HT29), breast (MCF-7, SKBR3) and ovarian (A2780, SKOV3) cancer cells. Ponatinib elicited primarily apoptosis, while JQ1 and dBET1 caused G0/G1 cell cycle arrest and very mild cell death. Phospho-FGFR and MYC, major targets of ponatinib and BET inhibitors, were downregulated after treatment with single drugs. Remarkably, ponatinib was found to sensitize cells to BET antagonists by enhancing apoptotic cell death, and this effect was associated with downregulation of MYC. In summary, our data shows that ponatinib sensitizes colon, breast, and ovarian cancer cells to BET bromodomain inhibitors. Further studies are warranted to determine the clinical value of this phenomenon.
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http://dx.doi.org/10.18632/oncotarget.25474DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5995173PMC
May 2018

Phenotyping and Target Expression Profiling of CD34/CD38 and CD34/CD38 Stem- and Progenitor cells in Acute Lymphoblastic Leukemia.

Neoplasia 2018 06 15;20(6):632-642. Epub 2018 May 15.

Ludwig Boltzmann Cluster Oncology, Medical University of Vienna, Waehringer Guertel 18-20, 1090 Vienna, Austria; Department of Internal Medicine I, Division of Hematology and Hemostaseology, Medical University of Vienna, Waehringer Guertel 18-20, 1090 Vienna, Austria. Electronic address:

Leukemic stem cells (LSCs) are an emerging target of curative anti-leukemia therapy. In acute lymphoblastic leukemia (ALL), LSCs frequently express CD34 and often lack CD38. However, little is known about markers and targets expressed in ALL LSCs. We have examined marker- and target expression profiles in CD34/CD38 LSCs in patients with Ph ALL (n = 22) and Ph ALL (n = 27) by multi-color flow cytometry and qPCR. ALL LSCs expressed CD19 (B4), CD44 (Pgp-1), CD123 (IL-3RA), and CD184 (CXCR4) in all patients tested. Moreover, in various subgroups of patients, LSCs also displayed CD20 (MS4A1) (10/41 = 24%), CD22 (12/20 = 60%), CD33 (Siglec-3) (20/48 = 42%), CD52 (CAMPATH-1) (17/40 = 43%), IL-1RAP (13/29 = 45%), and/or CD135 (FLT3) (4/20 = 20%). CD25 (IL-2RA) and CD26 (DPPIV) were expressed on LSCs in Ph ALL exhibiting BCR/ABL1, whereas in Ph ALL with BCR/ABL1, LSCs variably expressed CD25 but did not express CD26. In Ph ALL, CD34/CD38 LSCs expressed IL-1RAP in 6/18 patients (33%), but did not express CD25 or CD26. Normal stem cells stained negative for CD25, CD26 and IL-1RAP, and expressed only low amounts of CD52. In xenotransplantation experiments, CD34/CD38 and CD34/CD38 cells engrafted NSG mice after 12-20 weeks, and targeting with antibodies against CD33 and CD52 resulted in reduced engraftment. Together, LSCs in Ph and Ph ALL display unique marker- and target expression profiles. In Ph ALL with BCR/ABL1, the LSC-phenotype closely resembles the marker-profile of CD34/CD38 LSCs in chronic myeloid leukemia, confirming the close biologic relationship of these neoplasms. Targeting of LSCs with specific antibodies or related immunotherapies may facilitate LSC eradication in ALL.
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http://dx.doi.org/10.1016/j.neo.2018.04.004DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5994777PMC
June 2018

Controlling drug release from mesoporous silica through an amorphous, nanoconfined 1-tetradecanol layer.

Eur J Pharm Biopharm 2018 Jun 7;127:318-325. Epub 2018 Mar 7.

"Carol Davila" University of Medicine and Pharmacy, Biophysics and Cellular Biotechnology Dept., 8 Eroii Sanitari Blvd., Bucharest 050474, Romania; "Carol Davila" University of Medicine and Pharmacy, Excellence Research Center in Biophysics and Cellular Biotechnology, 8 Eroii Sanitari Blvd., Bucharest 050474, Romania.

Mesoporous silica materials are promising nano-carriers for drug delivery systems. Even though there are many strategies for controlling the drug release kinetics, these must be adapted through trial and error on a case-by-case basis. Here we explore the possibility of tailoring the release kinetics of hydrophilic, water soluble therapeutic agents from mesoporous silica through addition of a hydrophobic excipient, 1-tetradecanol. In vitro drug release experiments performed at 37 °C, in phosphate buffer solution (pH 7.4) show that the addition of tetradecanol yields slower drug release kinetics, which was correlated with the presence of a liquid fatty alcohol interfacial layer. The layer mass is 11-23 wt.% of the metoprolol-loaded silica sample, and it causes up to 1.6 times decrease of initial release rate with respect to materials without the fatty alcohol. This effect does not depend of carrier pore arrangement, being noticed for both hexagonal MCM-41 and cubic KIT-5 mesoporous silica. The toxicity of tetradecanol-containing materials was evaluated by formazan-based viability assay on Opossum kidney epithelial cell line, and no significant toxicity was observed.
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http://dx.doi.org/10.1016/j.ejpb.2018.02.020DOI Listing
June 2018

The KIT and PDGFRA switch-control inhibitor DCC-2618 blocks growth and survival of multiple neoplastic cell types in advanced mastocytosis.

Haematologica 2018 05 8;103(5):799-809. Epub 2018 Feb 8.

Ludwig Boltzmann Cluster Oncology, Medical University of Vienna, Austria

Systemic mastocytosis is a complex disease defined by abnormal growth and accumulation of neoplastic mast cells in various organs. Most patients exhibit a D816V-mutated variant of , which confers resistance against imatinib. Clinical problems in systemic mastocytosis arise from mediator-related symptoms and/or organ destruction caused by malignant expansion of neoplastic mast cells and/or other myeloid cells in various organ systems. DCC-2618 is a spectrum-selective pan KIT and PDGFRA inhibitor which blocks KIT D816V and multiple other kinase targets relevant to systemic mastocytosis. We found that DCC-2618 inhibits the proliferation and survival of various human mast cell lines (HMC-1, ROSA, MCPV-1) as well as primary neoplastic mast cells obtained from patients with advanced systemic mastocytosis (IC <1 μM). Moreover, DCC-2618 decreased growth and survival of primary neoplastic eosinophils obtained from patients with systemic mastocytosis or eosinophilic leukemia, leukemic monocytes obtained from patients with chronic myelomonocytic leukemia with or without concomitant systemic mastocytosis, and blast cells obtained from patients with acute myeloid leukemia. Furthermore, DCC-2618 was found to suppress the proliferation of endothelial cells, suggesting additional drug effects on systemic mastocytosis-related angiogenesis. Finally, DCC-2618 was found to downregulate IgE-mediated histamine release from basophils and tryptase release from mast cells. Together, DCC-2618 inhibits growth, survival and activation of multiple cell types relevant to advanced systemic mastocytosis. Whether DCC-2618 is effective in patients with advanced systemic mastocytosis is currently under investigation in clinical trials.
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http://dx.doi.org/10.3324/haematol.2017.179895DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5927976PMC
May 2018

Evaluation of cooperative antileukemic effects of nilotinib and vildagliptin in Ph chronic myeloid leukemia.

Exp Hematol 2018 01 12;57:50-59.e6. Epub 2017 Oct 12.

Ludwig Boltzmann Cluster Oncology, Medical University of Vienna, Vienna, Austria; Institute of Laboratory Animal Science, University of Veterinary Medicine Vienna, Vienna, Austria.

Chronic myeloid leukemia (CML) is a stem cell (SC) neoplasm characterized by the BCR/ABL1 oncogene. Although the disease can be kept under control using BCR/ABL1 tyrosine kinase inhibitors (TKIs) in most cases, some patients relapse or have resistant disease, so there is a need to identify new therapeutic targets in this malignancy. Recent data suggest that leukemic SCs (LSCs) in CML display the stem-cell (SC)-mobilizing cell surface enzyme dipeptidyl-peptidase IV (DPPIV = CD26) in an aberrant manner. In the present study, we analyzed the effects of the DPPIV blocker vildagliptin as single agent or in combination with the BCR/ABL1 TKI imatinib or nilotinib on growth and survival of CML LSCs in vitro and on LSC engraftment in an in vivo xenotransplantation nonobese diabetic SCID-IL-2Rγ (NSG) mouse model. We found that nilotinib induces apoptosis in CML LSCs and inhibits their engraftment in NSG mice. In contrast, no substantial effects were seen with imatinib or vildagliptin. Nevertheless, vildagliptin was found to reduce the "mobilization" of CML LSCs from a stroma cell layer consisting of mouse fibroblasts in an in vitro co-culture model, suggesting reduced disease expansion. However, although vildagliptin and nilotinib produced cooperative effects in individual experiments, overall, no significant effects of coadministered vildagliptin over nilotinib or imatinib treatment alone were seen on the engraftment of CML cells in NSG mice. Gliptins may be interesting drugs in the context of CML and nilotinib therapy, but our preclinical studies did not reveal a major cooperative effect of the drug-combination vildagliptin + nilotinib on engraftment of CML cells in NSG mice.
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http://dx.doi.org/10.1016/j.exphem.2017.09.012DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7115814PMC
January 2018

The pan-BCL-2-blocker obatoclax (GX15-070) and the PI3-kinase/mTOR-inhibitor BEZ235 produce cooperative growth-inhibitory effects in ALL cells.

Oncotarget 2017 Sep 28;8(40):67709-67722. Epub 2017 Jun 28.

Department of Internal Medicine I, Division of Hematology & Hemostaseology, Medical University of Vienna, Vienna, Austria.

Acute lymphoblastic leukemia (ALL) is characterized by leukemic expansion of lymphoid blasts in hematopoietic tissues. Despite improved therapy only a subset of patients can be cured. Therefore, current research is focusing on new drug-targets. Members of the BCL-2 family and components of the PI3-kinase/mTOR pathway are critically involved in the regulation of growth and survival of ALL cells. We examined the effects of the pan-BCL-2 blocker obatoclax and the PI3-kinase/mTOR-inhibitor BEZ235 on growth and survival of ALL cells. In H-thymidine uptake experiments, both drugs suppressed the proliferation of leukemic cells in all patients with Philadelphia chromosome-positive (Ph) ALL and Ph ALL (obatoclax IC: 0.01-5 μM; BEZ235, IC: 0.01-1 μM). Both drugs were also found to produce growth-inhibitory effects in all Ph and all Ph cell lines tested. Moreover, obatoclax and BEZ235 induced apoptosis in ALL cells. In drug-combination experiments, obatoclax and BEZ235 exerted synergistic growth-inhibitory effects on ALL cells. Finally, we confirmed that ALL cells, including CD34/CD38 stem cells and all cell lines express transcripts for PI3-kinase, mTOR, BCL-2, MCL-1, and BCL-xL. Taken together, this data shows that combined targeting of the PI3-kinase/mTOR-pathway and BCL-2 family-members is a potent approach to counteract growth and survival of ALL cells.
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http://dx.doi.org/10.18632/oncotarget.18810DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5620205PMC
September 2017

Combined targeting of STAT3 and STAT5: a novel approach to overcome drug resistance in chronic myeloid leukemia.

Haematologica 2017 09 8;102(9):1519-1529. Epub 2017 Jun 8.

Department of Internal Medicine I, Division of Hematology and Hemostaseology, Medical University of Vienna, Austria.

In chronic myeloid leukemia, resistance against BCR-ABL1 tyrosine kinase inhibitors can develop because of mutations, activation of additional pro-oncogenic pathways, and stem cell resistance. Drug combinations covering a broad range of targets may overcome resistance. CDDO-Me (bardoxolone methyl) is a drug that inhibits the survival of leukemic cells by targeting different pro-survival molecules, including STAT3. We found that CDDO-Me inhibits proliferation and survival of tyrosine kinase inhibitor-resistant cell lines and primary leukemic cells, including cells harboring or T315I compound mutations. Furthermore, CDDO-Me was found to block growth and survival of CD34/CD38 leukemic stem cells (LSC). Moreover, CDDO-Me was found to produce synergistic growth-inhibitory effects when combined with BCR-ABL1 tyrosine kinase inhibitors. These drug-combinations were found to block multiple signaling cascades and molecules, including STAT3 and STAT5. Furthermore, combined targeting of STAT3 and STAT5 by shRNA and STAT5-targeting drugs also resulted in synergistic growth-inhibition, pointing to a new efficient concept of combinatorial STAT3 and STAT5 inhibition. However, CDDO-Me was also found to increase the expression of heme-oxygenase-1, a heat-shock-protein that triggers drug resistance and cell survival. We therefore combined CDDO-Me with the heme-oxygenase-1 inhibitor SMA-ZnPP, which also resulted in synergistic growth-inhibitory effects. Moreover, SMA-ZnPP was found to sensitize cells against the combination 'CDDO-Me+ tyrosine kinase inhibitor'. Together, combined targeting of STAT3, STAT5, and heme-oxygenase-1 overcomes resistance in cells, including stem cells and highly resistant sub-clones expressing BCR-ABL1 or T315I-compound mutations. Whether such drug-combinations are effective in tyrosine kinase inhibitor-resistant patients with chronic myeloid leukemia remains to be elucidated.
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http://dx.doi.org/10.3324/haematol.2016.163436DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5685220PMC
September 2017

The JAK2/STAT5 signaling pathway as a potential therapeutic target in canine mastocytoma.

Vet Comp Oncol 2018 Mar 11;16(1):55-68. Epub 2017 Apr 11.

Department of Internal Medicine I, Division of Hematology and Hemostaseology, Medical University of Vienna, Vienna, Austria.

Background: Mastocytoma are frequently diagnosed cutaneous neoplasms in dogs. In non-resectable mastocytoma patients, novel targeted drugs are often applied. The transcription factor STAT5 has been implicated in the survival of human neoplastic mast cells (MC). Our study evaluated the JAK2/STAT5 pathway as a novel target in canine mastocytoma.

Materials And Methods: We employed inhibitors of JAK2 (R763, TG101348, AZD1480, ruxolitinib) and STAT5 (pimozide, piceatannol) and evaluated their effects on 2 mastocytoma cell lines, C2 and NI-1.

Results: Activated JAK2 and STAT5 were detected in both cell lines. The drugs applied were found to inhibit proliferation and survival in these cells with the following rank-order of potency: R763 > TG101348 > AZD1480 > pimozide > ruxolitinib > piceatannol. Moreover, synergistic anti-neoplastic effects were obtained by combining pimozide with KIT-targeting drugs (toceranib, masitinib, nilotinib, midostaurin) in NI-1 cells.

Conclusion: The JAK2/STAT5 pathway is a novel potential target of therapy in canine mastocytoma.
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http://dx.doi.org/10.1111/vco.12311DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5824979PMC
March 2018

Box-Behnken experimental design for chromium(VI) ions removal by bacterial cellulose-magnetite composites.

Int J Biol Macromol 2016 Oct 22;91:1062-72. Epub 2016 Jun 22.

University "Politehnica" of Bucharest, Faculty of Electrical Engineering, 313 Spl. Independentei Street, 060042 Bucharest, Romania.

In this study bacterial cellulose-magnetite composites were synthesised for the removal of chromium(VI) from aqueous solutions. Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), thermogravimetric analysis and X-ray Photoelectron Spectroscopy (XPS) were used to characterize the bacterial cellulose-magnetite composites and to reveal the uniform dispersion of nanomagnetite in the BC matrix. Magnetic properties were also measured to confirm the magnetite immobilization on bacterial cellulose membrane. The effects of initial Cr(VI) concentration, solution pH and solid/liquid ratio upon chromium removal were examined using the statistical Box-Behnken Design. Because of the possibility of magnetite dissolution during chromium(VI) adsorption, the degree of iron leaching was also analysed in the same conditions as Cr(VI) adsorption. From the factors affecting chromium(VI) adsorption the most important was solution pH. The highest Cr(VI) removal efficiency was observed at pH 4, accompanied by the lowest iron leaching in the solution. The adsorption experiments also indicated that the adsorption process of chromium(VI) is well described by Freundlich adsorption model. Our results proved that the BC-magnetite composites could be used for an efficient removal of chromium(VI) from diluted solutions with a minimum magnetite dissolution during operation.
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http://dx.doi.org/10.1016/j.ijbiomac.2016.06.070DOI Listing
October 2016

The Validation of a Spanish Version of the Multidimensional Inventory of Religious/Spiritual Well-Being in Mexican College Students.

Span J Psychol 2016 Feb 18;19:E3. Epub 2016 Feb 18.

University of Graz(Austria).

After the Multidimensional Inventory for Religious/Spiritual Well-Being (MI-RSWB) was validated as a reliable instrument for the Western European context it is primarily intended in this study to translate the measure into Spanish and adapt it for the Mexican culture. Furthermore we investigate whether spirituality/religiosity has a similar impact on indicators of personality and subjective well-being in Mexico as it does in samples drawn from Western European cultures. 190 students (99 females) from public and private universities in Guadalajara, all Mexican citizens, were involved in this study. We found strong evidential support for the six factor solution of the Original MI-RSWB in this Mexican population. By mirroring previous research the measure showed a highly satisfying internal consistency (α = .91 for the total score and .75 or higher for all six sub dimensions). Furthermore the total RSWB score was observed to be related with Eysenck's personality dimensions Extraversion (r = .24, p < .01), and Psychoticism (r = -.28, p < .001), although not with Neuroticism. There was also a positive correlation with Sense of Coherence (r = .31, p < .001). In conclusion, the dimensionality of RSWB and its associations with personality and subjective well-being was well supported in this first application within a Mexican cultural context.
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http://dx.doi.org/10.1017/sjp.2016.9DOI Listing
February 2016

Identification of CD25 as STAT5-Dependent Growth Regulator of Leukemic Stem Cells in Ph+ CML.

Clin Cancer Res 2016 Apr 25;22(8):2051-61. Epub 2015 Nov 25.

Division of Hematology and Hemostaseology, Department of Internal Medicine I, Medical University of Vienna, Vienna, Austria. Ludwig Boltzmann Cluster Oncology, Medical University of Vienna, Vienna, Austria.

Purpose: In chronic myelogenous leukemia (CML), leukemic stem cells (LSC) represent a critical target of therapy. However, little is known about markers and targets expressed by LSCs. The aim of this project was to identify novel relevant markers of CML LSCs.

Experimental Design: CML LSCs were examined by flow cytometry, qPCR, and various bioassays. In addition, we examined the multipotent CD25(+)CML cell line KU812.

Results: In contrast to normal hematopoietic stem cells, CD34(+)/CD38(-)CML LSCs expressed the IL-2 receptor alpha chain, IL-2RA (CD25). STAT5 was found to induce expression of CD25 in Lin(-)/Sca-1(+)/Kit(+)stem cells in C57Bl/6 mice. Correspondingly, shRNA-induced STAT5 depletion resulted in decreased CD25 expression in KU812 cells. Moreover, the BCR/ABL1 inhibitors nilotinib and ponatinib were found to decrease STAT5 activity and CD25 expression in KU812 cells and primary CML LSCs. A CD25-targeting shRNA was found to augment proliferation of KU812 cellsin vitroand their engraftmentin vivoin NOD/SCID-IL-2Rγ(-/-)mice. In drug-screening experiments, the PI3K/mTOR blocker BEZ235 promoted the expression of STAT5 and CD25 in CML cells. Finally, we found that BEZ235 produces synergistic antineoplastic effects on CML cells when applied in combination with nilotinib or ponatinib.

Conclusions: CD25 is a novel STAT5-dependent marker of CML LSCs and may be useful for LSC detection and LSC isolation in clinical practice and basic science. Moreover, CD25 serves as a growth regulator of CML LSCs, which may have biologic and clinical implications and may pave the way for the development of new more effective LSC-eradicating treatment strategies in CML.
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http://dx.doi.org/10.1158/1078-0432.CCR-15-0767DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4817228PMC
April 2016

Correlation of the intracellular reactive oxygen species levels with textural properties of functionalized mesostructured silica.

J Biomed Mater Res A 2014 Dec 25;102(12):4435-42. Epub 2014 Feb 25.

Department of Inorganic Chemistry, Physical Chemistry and Electrochemistry, University "Politehnica" of Bucharest, Bucharest, 011061, Romania.

Mesostructured silica is frequently used in biomedical applications, being considered nontoxic and biocompatible material, suitable for the development of drug delivery systems (DDS). Four functionalized MCM-41 silica materials with hydrophobic (methyl and vinyl) and hydrophilic (3-aminopropyl and 3-mercaptopropyl) groups were obtained by post-synthesis functionalization and characterized by small-angle X-ray diffraction, infrared spectroscopy, thermal analysis, and nitrogen adsorption-desorption isotherms. The main structural and textural parameters of the obtained silica were determined. The effect of the functionalized silica on fibroblast (NIH3T3) and melanocyte cells (B16F10) was studied with respect to the proliferation rate and the levels of reactive oxygen species (ROS). It was found that the textural properties of all samples influenced the levels of intracellular ROS and consequently, the proliferation rate. Both, healthy and malignant cells exhibited linear dependence of ROS levels with the specific surface area values, but with different response. The contribution of the methyl functionalized silica to the ROS level is apart to the general trend.
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http://dx.doi.org/10.1002/jbm.a.35131DOI Listing
December 2014

Type 1 fimbriae contribute to catheter-associated urinary tract infections caused by Escherichia coli.

J Bacteriol 2014 Mar 13;196(5):931-9. Epub 2013 Dec 13.

University of Applied Sciences, Biomedical Science, Graz, Austria.

Biofilm formation on catheters is thought to contribute to persistence of catheter-associated urinary tract infections (CAUTI), which represent the most frequent nosocomial infections. Knowledge of genetic factors for catheter colonization is limited, since their role has not been assessed using physicochemical conditions prevailing in a catheterized human bladder. The current study aimed to combine data from a dynamic catheterized bladder model in vitro with in vivo expression analysis for understanding molecular factors relevant for CAUTI caused by Escherichia coli. By application of the in vitro model that mirrors the physicochemical environment during human infection, we found that an E. coli K-12 mutant defective in type 1 fimbriae, but not isogenic mutants lacking flagella or antigen 43, was outcompeted by the wild-type strain during prolonged catheter colonization. The importance of type 1 fimbriae for catheter colonization was verified using a fimA mutant of uropathogenic E. coli strain CFT073 with human and artificial urine. Orientation of the invertible element (IE) controlling type 1 fimbrial expression in bacterial populations harvested from the colonized catheterized bladder in vitro suggested that the vast majority of catheter-colonizing cells (up to 88%) express type 1 fimbriae. Analysis of IE orientation in E. coli populations harvested from patient catheters revealed that a median level of ∼73% of cells from nine samples have switched on type 1 fimbrial expression. This study supports the utility of the dynamic catheterized bladder model for analyzing catheter colonization factors and highlights a role for type 1 fimbriae during CAUTI.
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http://dx.doi.org/10.1128/JB.00985-13DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3957706PMC
March 2014