Inhibition of Nitric Oxide and Prostaglandin E2 Expression by Methanol Extract of Polyopes affinis in Lipopolysaccharide-stimulated BV2 Microglial Cells through Suppression of Akt-dependent NF-kB Activity and MAPK Pathway

Purpose: To determine whether the methanol extract of Polyopes affinis (MEPA) down-regulates the expression of pro-inflammatory mediators in lipopolysaccharide (LPS)-stimulated BV2 microglial cells.Methods: The production of nitric oxide (NO) and prostaglandin E2 (PGE2) was measured by the Griess reagents and enzyme-linked immunosorbent assay (ELISA), respectively. Expression levels of mRNA and protein in LPS-stimulated BV2 microglial cells were assessed by reverse transcription-polymerase (RT-PCR) and Western blot analysis. Activation of nuclear factor-êB (NF-êB) was detected by electrophoretic mobility shift assay (EMSA).Results: MEPA inhibited the expression of LPS-induced pro-inflammatory mediators, NO and PGE2, as well as their respective genes, iNOS and COX-2, at both protein and mRNA levels, without any accompanying cytotoxicity. Moreover, treatment with MEPA significantly suppressed the LPS-induced DNA-binding activity of NF-êB, which is known as a main transcription factor for the regulation of proinflammatory genes, as well as the nuclear translocation of its subunit p65 and p50, by degrading IêBá.MEPA increased Akt dephosphorylation which leads to suppression of the DNA-binding activity of NF-kB in LPS-stimulated BV2 microglial cells and suppressed phosphorylation of ERK and JNK, which are involved in the mitogen-activated protein kinase (MAPK) signaling pathway for regulating proinflammatory genes.Conclusion: Our results indicate that MEPA down-regulates  pro-inflammatory mediators such as NO and PGE2 by suppressing Akt-dependent NF-êB activity as well as phosphorylation of ERK and JNK inLPS-stimulated BV2 microglial cells.Keywords: Polyopes affinis, Nitric oxide, Prostaglandin E2, Nuclear factor-k

Barcoded DNA-Tag Reporters for Multiplex Cis-Regulatory Analysis

Cis-regulatory DNA sequences causally mediate patterns of gene expression, but efficient experimental analysis of these control systems has remained challenging. Here we develop a new version of “barcoded" DNA-tag reporters, “Nanotags" that permit simultaneous quantitative analysis of up to 130 distinct cis-regulatory modules (CRMs). The activities of these reporters are measured in single experiments by the NanoString RNA counting method and other quantitative procedures. We demonstrate the efficiency of the Nanotag method by simultaneously measuring hourly temporal activities of 126 CRMs from 46 genes in the developing sea urchin embryo, otherwise a virtually impossible task. Nanotags are also used in gene perturbation experiments to reveal cis-regulatory responses of many CRMs at once. Nanotag methodology can be applied to many research areas, ranging from gene regulatory networks to functional and evolutionary genomics

Growth behavior of titanium dioxide thin films at different precursor temperatures

The hydrophilic TiO2 films were successfully deposited on slide glass substrates using titanium tetraisopropoxide as a single precursor without carriers or bubbling gases by a metal-organic chemical vapor deposition method. The TiO2 films were employed by scanning electron microscopy, Fourier transform infrared spectrometry, UV-Visible [UV-Vis] spectroscopy, X-ray diffraction, contact angle measurement, and atomic force microscopy. The temperature of the substrate was 500°C, and the temperatures of the precursor were kept at 75°C (sample A) and 60°C (sample B) during the TiO2 film growth. The TiO2 films were characterized by contact angle measurement and UV-Vis spectroscopy. Sample B has a very low contact angle of almost zero due to a superhydrophilic TiO2 surface, and transmittance is 76.85% at the range of 400 to 700 nm, so this condition is very optimal for hydrophilic TiO2 film deposition. However, when the temperature of the precursor is lower than 50°C or higher than 75°C, TiO2 could not be deposited on the substrate and a cloudy TiO2 film was formed due to the increase of surface roughness, respectively

The Association of State Opioid Misuse Prevention Policies With Patient‐ and Provider‐Related Outcomes: A Scoping Review

Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/154648/1/milq12436.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/154648/2/milq12436_am.pd

Model for performance prediction in multi-axis machining

This paper deals with a predictive model of kinematical performance in 5-axis milling within the context of High Speed Machining. Indeed, 5-axis high speed milling makes it possible to improve quality and productivity thanks to the degrees of freedom brought by the tool axis orientation. The tool axis orientation can be set efficiently in terms of productivity by considering kinematical constraints resulting from the set machine-tool/NC unit. Capacities of each axis as well as some NC unit functions can be expressed as limiting constraints. The proposed model relies on each axis displacement in the joint space of the machine-tool and predicts the most limiting axis for each trajectory segment. Thus, the calculation of the tool feedrate can be performed highlighting zones for which the programmed feedrate is not reached. This constitutes an indicator for trajectory optimization. The efficiency of the model is illustrated through examples. Finally, the model could be used for optimizing process planning

Detection of transgene in early developmental stage by GFP monitoring enhances the efficiency of genetic transformation of pepper

In order to establish a reliable and highly efficient method for genetic transformation of pepper, a monitoring system featuring GFP (green fluorescent protein) as a report marker was applied to Agrobacterium-mediated transformation. A callus-induced transformation (CIT) system was used to transform the GFP gene. GFP expression was observed in all tissues of T0, T1 and T2 peppers, constituting the first instance in which the whole pepper plant has exhibited GFP fluorescence. A total of 38 T0 peppers were obtained from 4,200 explants. The transformation rate ranged from 0.47 to 1.83% depending on the genotype, which was higher than that obtained by CIT without the GFP monitoring system. This technique could enhance selection power by monitoring GFP expression at the early stage of callus in vitro. The detection of GFP expression in the callus led to successful identification of the shoot that contained the transgene. Thus, this technique saved lots of time and money for conducting the genetic transformation process of pepper. In addition, a co-transformation technique was applied to the target transgene, CaCS (encoding capsaicinoid synthetase of Capsicum) along with GFP. Paprika varieties were transformed by the CaCS::GFP construct, and GFP expression in callus tissues of paprika was monitored to select the right transformant

Potassium channel gene mutations rarely cause atrial fibrillation

BACKGROUND: Mutations in several potassium channel subunits have been associated with rare forms of atrial fibrillation. In order to explore the role of potassium channels in inherited typical forms of the arrhythmia, we have screened a cohort of patients from a referral clinic for mutations in the channel subunit genes implicated in the arrhythmia. We sought to determine if mutations in KCNJ2 and KCNE1-5 are a common cause of atrial fibrillation. METHODS: Serial patients with lone atrial fibrillation or atrial fibrillation with hypertension were enrolled between June 1, 2001 and January 6, 2005. Each patient underwent a standardized interview and physical examination. An electrocardiogram, echocardiogram and blood sample for genetic analysis were also obtained. Patients with a family history of AF were screened for mutations in KCNJ2 and KCNE1-5 using automated sequencing. RESULTS: 96 patients with familial atrial fibrillation were enrolled. Eighty-three patients had lone atrial fibrillation and 13 had atrial fibrillation and hypertension. Patients had a mean age of 56 years at enrollment and 46 years at onset of atrial fibrillation. Eighty-one percent of patients had paroxysmal atrial fibrillation at enrollment. Unlike patients with an activating mutation in KCNQ1, the patients had a normal QT(c )interval with a mean of 412 ± 42 ms. Echocardiography revealed a normal mean ejection fraction of 62.0 ± 7.2 % and mean left atrial dimension of 39.9 ± 7.0 mm. A number of common polymorphisms in KCNJ2 and KCNE1-5 were identified, but no mutations were detected. CONCLUSION: Mutations in KCNJ2 and KCNE1-5 rarely cause typical atrial fibrillation in a referral clinic population

Transparent and flexible fingerprint sensor array with multiplexed detection of tactile pressure and skin temperature

We developed a transparent and flexible, capacitive fingerprint sensor array with multiplexed, simultaneous detection of tactile pressure and finger skin temperature for mobile smart devices. In our approach, networks of hybrid nanostructures using ultra-long metal nanofibers and finer nanowires were formed as transparent, flexible electrodes of a multifunctional sensor array. These sensors exhibited excellent optoelectronic properties and outstanding reliability against mechanical bending. This fingerprint sensor array has a high resolution with good transparency. This sensor offers a capacitance variation ~17 times better than the variation for the same sensor pattern using conventional ITO electrodes. This sensor with the hybrid electrode also operates at high frequencies with negligible degradation in its performance against various noise signals from mobile devices. Furthermore, this fingerprint sensor array can be integrated with all transparent forms of tactile pressure sensors and skin temperature sensors, to enable the detection of a finger pressing on the display