Effect of ALDH1A1 Gene Knockout on Drug Resistance in Paclitaxel and Topotecan Resistant Human Ovarian Cancer Cell Lines in 2D and 3D Model

Ovarian cancer is the most common cause of gynecological cancer death. Cancer Stem Cells (CSCs) characterized by drug transporters and extracellular matrix (ECM) molecules expression are responsible for drug resistance development. The goal of our study was to examine the role of aldehyde dehydrogenase 1A1 (ALDH1A1) expression in paclitaxel (PAC) and topotecan (TOP) resistant ovarian cancer cell lines. In both cell lines, we knocked out the ALDH1A1 gene using the CRISPR/Cas9 technique. Additionally, we derived an ALDH1A1 positive TOP-resistant cell line with ALDH1A1 expression in all cells via clonal selection. The effect of ALDH1A1 gene knockout or clonal selection on the expression of ALDH1A1, drug transporters (P-gp and BCRP), and ECM (COL3A1) was determined by Q-PCR, Western blot and immunofluorescence. Using MTT assay, we compared drug resistance in two-dimensional (2D) and three-dimensional (3D) cell culture conditions. We did not observe any effect of ALDH1A1 gene knockout on MDR1/P-gp expression and drug resistance in the PAC-resistant cell line. The knockout of ALDH1A1 in the TOP-resistant cell line resulted in a moderate decrease of BCRP and COL3A1 expression and weakened TOP resistance. The clonal selection of ALDH1A1 cells resulted in very strong downregulation of BCPR and COL3A1 expression and overexpression of MDR1/P-gp. This finally resulted in decreased resistance to TOP but increased resistance to PAC. All spheroids were more resistant than cells growing as monolayers, but the resistance mechanism differs. The spheroids’ resistance may result from the presence of cell zones with different proliferation paces, the density of the spheroid, ECM expression, and drug capacity to diffuse into the spheroid.     

Reaktionen des 2,6-Di-tert.-butyl-4-(N-tert.-butylnitrono)-phenoxyradikals

Reactions of the 2,6-Di-tert.-butyl-4-(N-tert.-butylnitrono)-phenoxyl Radical The title compound, a phenoxyl radical containing a nitrono group, reacts with alcohols and tert.-butylhydroperoxide yielding phenol and products of secondary radical solvent reactions. The reactions with lead tetraacetate, tert.-butoxy and 2-cyanoisopropyl radicals give high yields of cyclohexadienone adducts ( 6, 7 and 10 ) containing unchanged nitrono function. The reaction with dibenzoylperoxide, however, leads to the modification of the nitrono group yielding the N-benzoyloxycarboxamide ( 8 ). In the acidic decomposition of the tert.-butoxy radical adduct we suggest a nitrenium ion ( 16 ) as an intermediate.     

Syngas from sugarcane pyrolysis: An experimental study for fuel cell applications

The use of biomass for the production of electrical energy is a promising technological solution for those countries where there are problems with the disposal of agricultural waste and/or the production of low-cost energy. The gasification and/or pyrolysis of the biomass produces a gas rich in hydrogen that can be used in a fuel cell system to produce electrical energy with reduced environmental impact and significant energy recovery.In this work, a study of the pyrolysis of Brazilian sugarcane bagasse was carried out. The experimental process consisted of the pyrolysis of the biomass material in a batch pyrolysis reactor. In some runs the biomass was dry, while in others it was pre-treated by the addition of water. It was noted that the water added to the biomass before the pyrolysis process resulted in a decrease in the quantity of steam added to the fuel cell feeding gas, necessary to avoid carbon deposition, and in an increase in cell power, but, at the same time, caused a decrease in the quantity of syngas produced.Then, the composition of the gas obtained from the experimental pyrolysis of the sugarcane was inserted in a simulation tool of a molten carbonate fuel cell system in order to estimate the feasibility of the entire process in terms of operating conditions and electrical performance.The present study indicates that the syngas obtained from the sugarcane biomass (about 40%) can be converted into electricity using a fuel cell system with a high efficiency.     

Field determination of total dissolved inorganic carbon (TDIC) in natural waters using an IR analyzer: I. Preliminary laboratory tests

Preliminary laboratory tests carried out on 20 mineral waters indicate that the infrared (IR) method for the determination of total dissolved inorganic carbon (TDIC) is more accurate than the pH-alkalinity method, especially for high-TDIC, naturally sparkling waters and low-TDIC waters. The results of the two analytical methods are similar for mineral waters whose TDIC ranges from 100 to 500 mg/L as HCO₃, suggesting a relative accuracy of 0.3–2.9%. The limit of detection (LOD) of the IR method is close to 10 mg HCO₃/L and independent of pH. This value is comparable with the LOD of the pH-alkalinity method in the pH range 6.5–9 whereas the LOD of the latter increases strongly with decreasing pH below pH 6.5. The IR method is quick, since a run takes a few minutes and, on average, 10 analyses are carried out in an hour, including the time spent for clean up and preparation between samples. It constitutes a sound alternative to the pH-alkalinity technique for the field measurement of TDIC in geothermal and other natural waters.     

Changes of the cell surface hydrophobicity of Lactobacillus acidophilus La‐5 in response to pH,temperature and inulin

The main topic of this study was to study how cell surface hydrophobicity (CSH) could change in response to pH, temperature and inulin; Lactobacillus acidophilus La‐5 was used as a model microorganism. pH, temperature, inulin and incubation time (exposure to prebiotic or incubation at pH 4.0 and 9.0) were combined through a full factorial design and a Central Composite Design; the results were analysed using a multifactorial anova (first step) and a stepwise regression (second step). Temperature and pH significantly affected CSH: an increase in the temperature determined a significant increase in CSH, whereas the correlation pH vs. CSH was negative, as an increase in pH caused a significant decrease in CSH. Inulin played a significant role, but its effect could be influenced by temperature, pH and exposure time. This study is the first approach on the effects of some environmental factors on CSH and suggests that the culturing conditions and/or the exposure to some prebiotics could modify it with positive or negative effects.     

The Reciprocal Influences among Motivation,Personality Traits,and Game Habits for Playing Pokémon GO

This article reports a study exploring motivations of Pokémon Game use, individual differences related to personality traits, and game habits. First, it analyzed Pokémon GO motivations through exploratory factor analysis (EFA) by administering online the Pokémon GO Motivational Scale to a group of Italian gamers (N = 560). Successively, a Confirmatory Factor Analysis (CFA) was conducted testing three factorial models of Pokémon Game motivations on a selected random sample (N = 310). Results showed a three-factor model of Pokémon GO Game motivations (i.e. Personal Needs, Social Needs and Recreation), accounting for 68.9% of total variance plus a general higher order factor that best fits the data. Individual differences in Pokémon GO motivations and personality traits have been explored showing that high involved Pokémon GO players are introverted, low agreeableness, and conscientiousness people, driven by personal social and recreational needs. Reciprocal influences on motivational involvement, personality, and game habits were discussed.     

Optimized production of a high-performance hybrid biomaterial: biomineralized spider silk for bone tissue engineering

Silks have been widely used as biomaterials due to their biocompatibility, biodegradability, and excellent mechanical properties. In the present work, native spider silk was used as organic template for controlled nucleation of hydroxyapatite (HA) nano-coating that can act as biomimetic interface. Different bio-inspired neutralization methods and process parameters were evaluated to optimize the silk functionalization. The morphology and chemical composition were investigated by scanning electron microscopy, energy-dispersive X-ray spectroscopy, Fourier transform infrared spectroscopy, and X-ray diffraction analysis and mechanical properties were studied through tensile tests. Results showed that the optimized protocol enabled a controlled and homogeneous nucleation of apatite nano-crystals while maintaining silk mechanical performances after mineralization. This study reports the optimization of a simple and effective bio-inspired nucleation process for precise nucleation of HA onto spider silk templates, suitable to develop high-performance hybrid interfaces for bone tissue engineering. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2020 , 137, 48739.     

Aromatic Linkers Unleash the Antiproliferative Potential of 3-Chloropiperidines Against Pancreatic Cancer Cells

In this study, we describe the synthesis and biological evaluation of a set of bis-3-chloropiperidines (B−CePs) containing rigid aromatic linker structures. A modification of the synthetic strategy also enabled the synthesis of a pilot tris-3-chloropiperidine (Tri-CeP) bearing three reactive meta-chloropiperidine moieties on the aromatic scaffold. A structure–reactivity relationship analysis of B−CePs suggests that the arrangement of the reactive units affects the DNA alkylating activity, while also revealing correlations between the electron density of the aromatic system and the reactivity with biologically relevant nucleophiles, both on isolated DNA and in cancer cells. Interestingly, all aromatic 3-chloropiperidines exhibited a marked cytotoxicity and tropism for 2D and 3D cultures of pancreatic cancer cells. Therefore, the new aromatic 3-chloropiperidines appear to be promising contenders for further development of mustard-based anticancer agents aimed at pancreatic cancers.     

Emissions of VOCs,SVOCs, and mold during the construction process: Contribution to indoor air quality and future occupants’ exposure

Building materials and human activities are important sources of contamination indoors, but little information is available regarding contamination during construction process which could persist during the whole life of buildings. In this study, six construction stages on two construction sites were investigated regarding the emissions of 43 volatile organic compounds (VOCs), 46 semi-volatile organic compounds (SVOCs), and the presence of 4 genera of mold. Results show that the future indoor air quality does not only depend on the emissions of each building product but that it is also closely related to the whole implementation process. Mold spore measurements can reach 1400 CFU/m³, which is particularly high compared with the concentrations usually measured in indoor environments. Relatively low concentrations of VOCs were observed, in relation to the use of low emissive materials. Among SVOCs analyzed, some phthalates, permethrin, and hydrocarbons were found in significant concentrations upon the delivery of building as well as triclosan, suspected to be endocrine disruptor, and yet prohibited in the treatment of materials and construction since 2014. As some regulations exist for VOC emissions, it is necessary to implement them for SVOCs due to their toxicity.