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.     

ICA1L Is Associated with Small Vessel Disease: A Proteome-Wide Association Study in Small Vessel Stroke and Intracerebral Haemorrhage

Small vessel strokes (SVS) and intracerebral haemorrhages (ICH) are acute outcomes of cerebral small vessel disease (SVD). Genetic studies combining both phenotypes have identified three loci associated with both traits. However, the genetic cis-regulation at the protein level associated with SVD has not been studied before. We performed a proteome-wide association study (PWAS) using FUSION to integrate a genome-wide association study (GWAS) and brain proteomic data to discover the common mechanisms regulating both SVS and ICH. Dorsolateral prefrontal cortex (dPFC) brain proteomes from the ROS/MAP study (N = 376 subjects and 1443 proteins) and the summary statistics for the SVS GWAS from the MEGASTROKE study (N = 237,511) and multi-trait analysis of GWAS (MTAG)-ICH–SVS from Chung et al. (N = 240,269) were selected. We performed PWAS and then a co-localization analysis with COLOC. The significant and nominal results were validated using a replication dPFC proteome (N = 152). The replicated results (q-value < 0.05) were further investigated for the causality relationship using summary data-based Mendelian randomization (SMR). One protein (ICA1L) was significantly associated with SVS (z-score = −4.42 and p-value = 9.6 × 10⁻⁶) and non-lobar ICH (z-score = −4.8 and p-value = 1.58 × 10⁻⁶) in the discovery PWAS, with a high co-localization posterior probability of 4. In the validation PWAS, ICA1L remained significantly associated with both traits. The SMR results for ICA1L indicated a causal association of protein expression levels in the brain with SVS (p-value = 3.66 × 10⁻⁵) and non-lobar ICH (p-value = 1.81 × 10⁻⁵). Our results show that the association of ICA1L with SVS and non-lobar ICH is conditioned by the cis-regulation of its protein levels in the brain.     

Fast Separation of Capsaicinoids from Peppers by Reversed Phase Ultra-Performance Liquid Chromatography: Comparation with Traditional High-Performance Liquid Chromatography Methods

A new chromatographic method for the separation of major capsaicinoids in peppers has been developed. Nordihydrocapsaicin, capsaicin, dihydrocapsaicin, homocapsaicin, and homodihydrocapsaicin have been separated by reversed-phase ultra-performance liquid chromatography. A gradient method has been developed using two solvents: 0.1% acetic acid in water and 0.1% acetic acid in methanol. The developed method allows the full separation of capsaicinoids in less than 3 min, with high reproducibility (relative standard deviation < 4.3%) and repeatability (relative standard deviation < 3.6%). Robustness regarding the total amount of methanol in the sample was determined. Comparison with previous reversed-phase high-performance liquid chromatography methods using both monolithic and conventional columns was also studied. Finally, the method was applied in the determination of major capsaicinoids in 16 hot pepper samples produced in Spain.     

Passive adaptation or active engagement? The challenges of Housing First internationally and in the Italian case

In recent years, a peculiar homelessness policy that goes under the name of ‘Housing First’ has become increasingly popular all over the world. Epitomising a quintessential case of policy-mobility, Housing First can today be considered an heterogeneous assemblage of experiences and approaches that sometimes have little in common with each other. Introducing and commenting upon this heterogeneity, the paper critically analyses why and how Housing First has become a planetary success and what are the issues at stake with its widespread implementation. If recent scholarship published in this journal has granted us a fine understanding of Housing First’s functioning in the US, this paper offers something currently absent from the debate: a nuanced and critical understanding of the ambiguities related to the international success of this policy, with specific references to the challenges associated to its translation in the Italian case.     

Difference equation matrix model (DEMM) for the control of wind turbines

In this paper, a new algorithm, difference equation matrix model (DEMM), in the framework of model predictive control (MPC) is introduced. Instead of the standard dynamic matrix control (DMC), which is based upon step response method that has been used in most research works, we propose a new approach based upon a DEMM for model prediction. It has shown that DEMM has proven to be less computational and thus faster than the original DMC for real time applications. Thus, the drawbacks of DMC for online identification or adaptive design could be avoided. The control of wind turbines is carried out in order to decrease the cost of wind energy by increasing the efficiency, and thus the energy capture, or by reducing structural loading and increasing the lifetimes of the components and turbine structures. Modeling of wind turbine has been carried out. Effect of noise and disturbance on the system has been also studied. The results obtained show that the proposed DEMM minimizes the effect of the disturbance and produces an accurate and smooth control. Significant improvements in the regulation of rotor speed at high wind speeds are obtained from the proposed DEMM, where control set points are obtained ahead of the disturbance, saving the turbine of the negative effects of them and thus increasing its lifetime. Copyright © 2012 John Wiley & Sons, Ltd.     

Influence of caring youth sport contexts on efficacy-related beliefs and social behaviors.

Understanding what factors influence positive youth development has been advocated by youth development researchers (P. L. Benson, 2006; J. S. Eccles & J. A. Gootman, 2002). Consequently, the purpose of this study was to examine whether perceptions of a caring youth sport context influenced prosocial and antisocial behavior through efficacy-related beliefs, that is, positive and negative affective self-regulatory efficacy (ASRE) and empathic self-efficacy (ESE). Multiethnic youths taking part in summer sport programs (N = 395) completed a questionnaire that measured perceptions of the caring climate, ESE, ASRE, and social behavior. Structural equation modeling was used to test whether ASRE and ESE mediated the relationship between caring and social behaviors. Findings revealed that perceptions of caring positively predicted ASRE and ESE. In turn, positive ASRE positively predicted ESE. Prosocial behaviors were positively linked to ESE, whereas antisocial behaviors were negatively predicted by positive ASRE. The results suggest that caring influences prosocial and antisocial behavior because such contexts develop youths' ability to monitor, manage, and control positive affect, which in turn enhances their belief in their ability to empathize. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Real-time heating and power characterization of cells in standard cell designs

In today's digital electronic integrated circuits device heating is one of the most critical issues. Overheating can cause failures in functionality and device malfunction. In certain circumstances overheating of ICs can cause physical destruction of the device itself. This paper introduces a solution to determine cell and gate heating curves across the standard cell IC's surface. The presented methodology and toolset is tightly integrated into standardized logic simulator engines thus providing digital circuit designers a low-level, cell-resolution temperature distribution map during logic simulations. Actual temperatures of each consisting cell of the design can be monitored throughout the whole logic simulation. By being able to monitor temperatures of digital cells during initial simulations, it allows us to detect hot-spots and overheating caused malfunctions far before manufacture. By using the spatial location and temperature magnitude of hot-spots acquired from the presented methodology, place and route (P&R) tools can be driven to change cell placement and routing in order to avoid heating caused failures. Additionally, cooling solutions can be developed using the simulated temperature maps of the IC's surface. This paper also presents various aspects of power characterization methods which were used throughout the experiments.     

Segmenting visitors of cultural events by motivation: A sequential non-linear clustering analysis of Italian Christmas Market visitors

Considering the importance of market segmentation as a marketing tool to determine promotional policies, this paper aims to contribute to the tourism literature using the two-level approach proposed by Vesanto and Alhoniemi (2000) as an alternative and effective method to conduct cluster analyses.For this purpose, an empirical study was conducted interviewing tourists who visited three different Christmas Markets in Northern Italy. The two-level approach is based on two clustering techniques used in sequence: a Self-Organizing Map (SOM) followed by a clustering algorithm. The Silhouette index (Rousseeuw, 1987) is used as a guideline during the second level in the selection process of both the best clustering techniques (between hierarchical and non-hierarchical) and the best partition.The analysis identified three cluster segments and this paper demonstrates the suitability of the clustering method adopted. In the discussion of the results, marketing and managerial implications are also highlighted.     

Pulsed laser induced birefringence switching in a biopolymer matrix containing azo-dye molecules

All optical switching has been studied using the Optical Kerr Effect (OKE) configuration in a biopolymer matrix containing an azo-dye: the Disperse Orange 3 (DO3). The biopolymer system consisted of a deoxyribonucleic acid blended with cationic surfactant molecule cetyltrimethyl-ammonium chloride is suitable for optical quality thin film fabrication. The excitation beams inducing birefringence were delivered from a continuous wave laser at 532 nm and another nanosecond pulsed Nd: YAG laser. The birefringence was instantaneously monitored under crossed polarizer system by a weak non-absorbed light from a cw He-Ne laser working at 632.8 nm. Fast all optical switching process (in the range of microseconds) and excellent reversibility have been observed.     

Numerical analysis of the width of fracture process zone in concrete beams

There are two ways of modelling cracking in concrete structures using a finite element method (FEM). In the first concept crack is considered as densely distributed throughout the finite area of element. The alternative approach assumes an isolated sharp interelement crack. The first concept of microcracked band is often used during numerical computations of concrete structures and a crack band model can be applied in design practice where relatively simple methods of calculations are preferred. In this model the width of microcracked zone, which is called the width of fracture process zone, is the additional parameter of tensile concrete.In this paper, the influence of the fracture process zone width on numerical calculations in case of bent concrete beams is analyzed. The results of numerical simulations are presented and compared with those obtained during the experiment. The crack band model proposed by Hillerborg [Cement and Concrete Research, 6 (1976) 773–782] has been used in numerical calculations and six different widths of fracture process zone have been modelled in the analyzed specimen. When comparing the obtained FEM-results their significant differences according to the width of fracture process zone have been observed. On the basis of the performed analysis it may be concluded that the width of fracture process zone taken in numerical simulations does have an influence on FEM-results.