Aspirin Inhibits the Inflammatory Response of Protease-Activated Receptor 1 in Pregnancy Neutrophils: Implications for Treating Women with Preeclampsia

Neutrophils expressing cyclooxygenase-2 (COX-2) extensively infiltrate maternal blood vessels in preeclampsia, associated with vascular inflammation. Because pregnancy neutrophils also express protease-activated receptor 1 (PAR-1, F2R thrombin receptor), which they do not in non-pregnant subjects, they can be activated by proteases. We tested the hypothesis that aspirin at a dose sufficient to inhibit COX-2 would reduce inflammatory responses in preeclampsia neutrophils. Neutrophils were isolated from normal pregnant and preeclamptic women at approximately 30 weeks’ gestation. Normal pregnancy neutrophils were treated with elastase, a protease elevated in preeclampsia, or elastase plus aspirin to inhibit COX-2, or elastase plus pinane thromboxane, a biologically active structural analog of thromboxane and a thromboxane synthase inhibitor. Preeclamptic pregnancy neutrophils were treated with the same doses of aspirin or pinane thromboxane. Confocal microscopy with immunofluorescence staining was used to determine the cellular localization of the p65 subunit of nuclear factor-kappa B (NF-κB) and media concentrations of thromboxane were measured to evaluate the inflammatory response. In untreated neutrophils of normal pregnant women, p65 was localized to the cytosol. Upon stimulation with elastase, p65 translocated from the cytosol to the nucleus coincident with increased thromboxane production. When neutrophils were co-treated with aspirin or pinane thromboxane, elastase was not able to cause nuclear translocation of p65 or increase thromboxane. In untreated neutrophils of preeclamptic women, the p65 subunit was present in the nucleus and thromboxane production was elevated, but when preeclamptic neutrophils were treated with aspirin or pinane thromboxane, p65 was cleared from the nucleus and returned to the cytosol along with decreased thromboxane production. These findings suggest that COX-2 is a downstream mediator of PAR-1 and demonstrate that PAR-1- mediated inflammation can be inhibited by aspirin. Given the extensive and ubiquitous expression of PAR-1 and COX-2 in preeclamptic women, consideration should be given to treating women with preeclampsia using a dose of aspirin sufficient to inhibit COX-2.     

ONT-Based Alternative Assemblies Impact on the Annotations of Unique versus Repetitive Features in the Genome of a Romanian Strain of Drosophila melanogaster

To date, different strategies of whole-genome sequencing (WGS) have been developed in order to understand the genome structure and functions. However, the analysis of genomic sequences obtained from natural populations is challenging and the biological interpretation of sequencing data remains the main issue. The MinION device developed by Oxford Nanopore Technologies (ONT) is able to generate long reads with minimal costs and time requirements. These valuable assets qualify it as a suitable method for performing WGS, especially in small laboratories. The long reads resulted using this sequencing approach can cover large structural variants and repetitive sequences commonly present in the genomes of eukaryotes. Using MinION, we performed two WGS assessments of a Romanian local strain of Drosophila melanogaster, referred to as Horezu_LaPeri (Horezu). In total, 1,317,857 reads with a size of 8.9 gigabytes (Gb) were generated. Canu and Flye de novo assembly tools were employed to obtain four distinct assemblies with both unfiltered and filtered reads, achieving maximum reference genome coverages of 94.8% (Canu) and 91.4% (Flye). In order to test the quality of these assemblies, we performed a two-step evaluation. Firstly, we considered the BUSCO scores and inquired for a supplemental set of genes using BLAST. Subsequently, we appraised the total content of natural transposons (NTs) relative to the reference genome (ISO1 strain) and mapped the mdg1 retroelement as a resolution assayer. Our results reveal that filtered data provide only slightly enhanced results when considering genes identification, but the use of unfiltered data had a consistent positive impact on the global evaluation of the NTs content. Our comparative studies also revealed differences between Flye and Canu assemblies regarding the annotation of unique versus repetitive genomic features. In our hands, Flye proved to be moderately better for gene identification, while Canu clearly outperformed Flye for NTs analysis. Data concerning the NTs content were compared to those obtained with ONT for the D. melanogaster ISO1 strain, revealing that our strategy conducted to better results. Additionally, the parameters of our ONT reads and assemblies are similar to those reported for ONT experiments performed on various model organisms, revealing that our assembly data are appropriate for a proficient annotation of the Horezu genome.     

Estimation of SPIO Nanoparticles Uptakes by Macrophages Using Transmission Electron Microscopy

Due to their interesting size-dependent magnetic characteristics and relative biocompatibility, magnetic superparamagnetic iron oxide (SPIO) nanoparticles have been widely exploited as probes for cell and subcellular structure identification, as well as medication and gene delivery. A thorough understanding of the mechanics of the interaction between nanoparticles and macrophages is vital in managing dynamic processes in nanomedicine. In this study, the interaction behavior and uptake of SPIO nanoparticles by M1- and M2-type macrophages were investigated. Mice monocytes were differentiated into M1 and M2 macrophages, and the uptake of SPIO nanoparticles was studied using a TEM microscope. A high resolution image of 1 nm resolution, an image processing technique, was developed to extract the SPIO-NPs from tomographic TEM microscopic images. Lysosomes appear to be the zones of high concentrations of SPIO inside macrophages. Lysosomes were first selected in each image, and then segmentation by the Otsu thresholding method was used to extract the SPIO-NPs. The Otsu threshold method is a global thresholding technique used to automatically differentiate SPIOs from the background. The SPIO-NPs appear in red colors, and the other pixels in the image are considered background. Then, an estimation of the SPIO-NP uptakes by lysosomes is produced. Higher uptake of all-sized nanoparticles was observed in M1- and M2-type macrophages. An accurate estimation of the number of SPIO-NPs was obtained. This result will help in controlling targeted drug delivery and assessing the safety impact of the use of SPIO-NPs in nanomedicine for humans.     

The Implications of Noncoding RNAs in the Evolution and Progression of Nonalcoholic Fatty Liver Disease (NAFLD)-Related HCC

Nonalcoholic fatty liver disease (NAFLD) is the most prevalent liver pathology worldwide. Meanwhile, liver cancer represents the sixth most common malignancy, with hepatocellular carcinoma (HCC) as the primary, most prevalent subtype. Due to the rising incidence of metabolic disorders, NAFLD has become one of the main contributing factors to HCC development. However, although NAFLD might account for about a fourth of HCC cases, there is currently a significant gap in HCC surveillance protocols regarding noncirrhotic NAFLD patients, so the majority of NAFLD-related HCC cases were diagnosed in late stages when survival chances are minimal. However, in the past decade, the focus in cancer genomics has shifted towards the noncoding part of the genome, especially on the microRNAs (miRNAs) and long noncoding RNAs (lncRNAs), which have proved to be involved in the regulation of several malignant processes. This review aims to summarize the current knowledge regarding some of the main dysregulated, noncoding RNAs (ncRNAs) and their implications for NAFLD and HCC development. A central focus of the review is on miRNA and lncRNAs that can influence the progression of NAFLD towards HCC and how they can be used as potential screening tools and future therapeutic targets.     

Bacillus subtilis: As an Efficient Bacterial Strain for the Reclamation of Water Loaded with Textile Azo Dye,Orange II

The azo dye orange II is used extensively in the textile sector for coloring fabrics. High concentrations of it are released into aqueous environments through textile effluents. Therefore, its removal from textile wastewater and effluents is necessary. Herein, initially, we tested 11 bacterial strains for their capabilities in the degradation of orange II dye. It was revealed in the preliminary data that B. subtilis can more potently degrade the selected dye, which was thus used in the subsequent experiments. To achieve maximum decolorization, the experimental conditions were optimized whereby maximum degradation was achieved at: a 25 ppm dye concentration, pH 7, a temperature of 35 °C, a 1000 mg/L concentration of glucose, a 1000 mg/L urea concentration, a 666.66 mg/L NaCl concentration, an incubation period of 3 days, and with hydroquinone as a redox mediator at a concentration of 66.66 mg/L. The effects of the interaction of the operational factors were further confirmed using response surface methodology, which revealed that at optimum conditions of pH 6.45, a dye concentration of 17.07 mg/L, and an incubation time of 9.96 h at 45.38 °C, the maximum degradation of orange II can be obtained at a desirability coefficient of 1, estimated using the central composite design (CCD). To understand the underlying principles of degradation of the metabolites in the aliquot mixture at the optimized condition, the study steps were extracted and analyzed using GC-MS(Gas Chromatography Mass Spectrometry), FTIR(Fourier Transform Infrared Spectroscopy), ¹H and carbon 13 NMR(Nuclear Magnetic Resonance Spectroscopy). The GC-MS pattern revealed that the original dye was degraded into o-xylene and naphthalene. Naphthalene was even obtained in a pure state through silica gel column isolation and confirmed using ¹H and ¹³C NMR spectroscopic analysis. Phytotoxicity tests on Vigna radiata were also conducted and the results confirmed that the dye metabolites were less toxic than the parent dye. These results emphasize that B. subtilis should be used as a potential strain for the bioremediation of textile effluents containing orange II and other toxic azo dyes.     

Cloaking a receiving antenna or a sensor with plasmonic metamaterials

In this contribution, we discuss the possibility of applying the scattering-cancellation-based plasmonic cloaking technique to reduce the scattering from a sensing device, lowering its disturbance on sensing without affecting its ability to measure and “sense” the external world. The plasmonic cloak, while allowing the external fields to penetrate into the cover, may be able to suppress the scattering without necessarily shielding its interiors from the impinging light, which may be of great use if we wish to “sense” the external world, or receive an external signal, from inside the cloak. Our results may provide a unique sensing system that may receive and transmit signals, while its presence may not be perceived by the surrounding. This may be of great importance in a wide range of biological, optical, physical and engineering applications.     

Thermal inactivation of Campylobacter jejuni in broth

New Zealand has a high rate of reported campylobacteriosis compared with other developed countries. One possible reason is that local strains have greater heat tolerance and thus are better able to survive undercooking; this hypothesis is supported by the remarkably high D-values reported for Campylobacter jejuni in The Netherlands. The objective of this study was to investigate the thermal inactivation of isolates from New Zealand in broth, using strains that are commonly found in human cases and food samples in New Zealand. Typed Campylobacter strains were heated to a predetermined temperature using a submerged-coil heating apparatus. The first-order kinetic model has been used extensively in the calculation of the thermal inactivation parameters, D and z; however, nonlinear survival curves have been reported, and a number of models have been proposed to describe the patterns observed. Therefore, this study compared the conventional first-order model with eight nonlinear models for survival curves. Kinetic parameters were estimated using both one- and two-step regression techniques. In general, nonlinear models fit the individual inactivation data sets better than the log-linear model. However, the log-linear and the (nonlinear) Weibull models were the only models that could be successfully fitted to all data sets. For seven relevant New Zealand C. jejuni strains, at temperatures from 51.5 to 60°C, D- and z-values were obtained, ranging from 1.5 to 228 s and 4 to 5.2°C, respectively. These values are in broad agreement with published international data and do not indicate that the studied New Zealand C. jejuni strains are more heat resistant than other strains, in contrast with some reports from The Netherlands.     

A low‐power,area‐efficient multichannel receiver for micro MRI

A new integrated, low‐noise, low‐power, and area‐efficient multichannel receiver for magnetic resonance imaging (MRI) is described. The proposed receiver presents an alternative technique to overcome the use of multiple receiver front‐ends in parallel MRI. The receiver consists of three main stages: low‐noise pre‐amplifier, quadrature down‐converter, and a band pass filter (BPF). These components are used to receive the nuclear magnetic resonance signals from a 3 × 3 array of micro coils. These signals are combined using frequency domain multiplexing (FDM) method in the pre‐amplifier and BPF stages, then amplified and filtered to remove any out‐of‐band noise before providing it to an analog‐to‐digital converter at the low intermediate frequency stage. The receiver is designed using a 90 nm CMOS technology to operate at the main B₀ magnetic field of 9.4 T, which corresponds to 400 MHz. The receiver has an input referred noise voltage of 1.1 nV/√Hz, a total voltage gain of 87 dB, a power consumption of 69 mA from a 1 V supply voltage, and an area of 305 µm × 530 µm including the reference current and bias voltage circuits. Copyright © 2013 John Wiley & Sons, Ltd.     

Energy-efficient operation of indirect adiabatic data center cooling systems via Newton-like phasor extremum seeking control

This paper considers the problem of optimizing the operation of Indirect Adiabatic Cooling (IAC) systems with application to data centers. Optimal operation is achieved when the required cooling demand is satisfied at the minimum energy cost. For this purpose, we design a supervisory control system, where the higher layer determines the optimal set-points for the local controllers by employing an Extremum Seeking Control (ESC) scheme. In particular, we consider a Newton-like phasor ESC, which augments the derivative estimator underlying the phasor approach to capture also the Hessian of the plant index and then it uses these estimates to steer the system along a Newton-like direction. The effectiveness of the considered approach is tested in simulation by exploiting a Matlab-based simulation environment including an IAC system and a computer room.