The development of microarrays permits us to monitor transcriptomes on a genome-wide scale. To validate microarray measurements, quantitative-real time-reverse transcription PCR (Q-RT-PCR) is one of ...the most robust and commonly used approaches. The new challenge in gene quantification analysis is how to explicitly incorporate statistical estimation in such studies. In the realm of statistical analysis, the various available methods of the probe level normalization for microarray analysis may result in distinctly different target selections and variation in the scores for the correlation between microarray and Q-RT-PCR. Moreover, it remains a major challenge to identify a proper internal control for Q-RT-PCR when confirming microarray measurements.
Sixty-six Affymetrix microarray slides using lung adenocarcinoma tissue RNAs were analyzed by a statistical re-sampling method in order to detect genes with minimal variation in gene expression. By this approach, we identified DDX5 as a novel internal control for Q-RT-PCR. Twenty-three genes, which were differentially expressed between adjacent normal and tumor samples, were selected and analyzed using 24 paired lung adenocarcinoma samples by Q-RT-PCR using two internal controls, DDX5 and GAPDH. The percentage correlation between Q-RT-PCR and microarray were 70% and 48% by using DDX5 and GAPDH as internal controls, respectively.
Together, these quantification strategies for Q-RT-PCR data processing procedure, which focused on minimal variation, ought to significantly facilitate internal control evaluation and selection for Q-RT-PCR when corroborating microarray data.
Honeysuckle (
Thunb) is a traditional Chinese medicine (TCM) with an antipathogenic activity. MicroRNAs (miRNAs) are small non-coding RNA molecules that are ubiquitously expressed in cells. ...Endogenous miRNA may function as an innate response to block pathogen invasion. The miRNA expression profiles of both mice and humans after the ingestion of honeysuckle were obtained. Fifteen overexpressed miRNAs overlapped and were predicted to be capable of targeting three viruses: dengue virus (DENV), enterovirus 71 (EV71) and SARS-CoV-2. Among them,
was examined to be capable of targeting the EV71 RNA genome by reporter assay and Western blotting. Moreover, honeysuckle-induced
suppression of EV71 RNA and protein expression as well as viral replication were investigated both in vitro and in vivo. We demonstrated that
targeted EV71 at the predicted sequences using luciferase reporter plasmids as well as two infectious replicons (pMP4-y-5 and pTOPO-4643). The suppression of EV71 replication and viral load was demonstrated in two cell lines by luciferase activity, RT-PCR, real-time PCR, Western blotting and plaque assay. Furthermore, EV71-infected suckling mice fed honeysuckle extract or inoculated with
showed decreased clinical scores and a prolonged survival time accompanied with decreased viral RNA, protein expression and virus titer. The ingestion of honeysuckle attenuates EV71 replication and related pathogenesis partially through the upregulation of
expression both in vitro and in vivo. Our previous report and the current findings imply that both honeysuckle and upregulated
can execute a suppressive function against the replication of DENV and EV71. Taken together, this evidence indicates that honeysuckle can induce the expression of
and that this miRNA as well as 11 other miRNAs have great potential to prevent and suppress EV71 replication.
A new approach by investigating the intra-tumoral microbiome raised great interest because they may influence the host immune response and natural history of the disease. However, previous studies on ...the intra-tumoral microbiome of pancreatic ductal adenocarcinoma (PDAC) were mostly based on examining the formalin-fixed paraffin-embedded tumor specimens. This study aims to investigate the feasibility of using endoscopic ultrasound-guided fine-needle biopsy (EUS-FNB) as a complementary procedure of surgical biopsy to obtain adequate fresh pancreatic cancer tissue for intra-tumoral microbial research. This was a prospective pilot study performed at a single tertiary referral center. We obtained pancreatic cancer tissue by EUS-FNB and surgical biopsy, respectively. We amplified the V3-V4 hyper-variable region of bacterial 16S ribosomal ribonucleic acid (rRNA) genes, constructed a pair-end library, and performed high-throughput sequencing. From August 2020 to November 2020, nine eligible patients with PDAC were enrolled in this study. The intra-tumoral microbiome profile was successfully generated from the PDAC cancer tissue obtained by EUS-FNB as well as by surgical biopsy. There was no significant difference in intra-tumoral alpha-diversity or bacterial taxonomic composition between tissues obtained by EUS-FNB and by surgical biopsy. EUS-FNB can collect sufficient fresh cancer tissue for microbiome analyses without complication. The intra-tumoral microbiome profile in tissues obtained by EUS-FNB had similar alpha-diversity and taxonomic profiles with those obtained by surgical biopsy. It implicated, except for surgical biopsy, EUS-FNB can be another valid and valuable tool for studying intra-tumoral microbiome in patients with resectable and unresectable PDAC.
Serological responses (Seroresponse) and durability of hepatitis A virus (HAV) vaccination are reduced among human immunodeficiency virus (HIV)‐positive patients. Incidence of and associated factors ...with early seroreversion (loss of seroresponse) among HIV‐positive patients who have achieved seroresponses after two doses of HAV vaccination remain unclear. In this multicenter study, we followed HIV‐positive adults who had mounted seroresponses after completing two doses of HAV vaccination during a recent outbreak of acute hepatitis A between 2015 and 2017, a 1:4 case‐control study was conducted to identify factors associated with seroreversion. Case patients were those with seroreversion, and controls were those with similar follow‐up durations who were able to maintain seroresponses. During the study period, 49 of the 1,256 patients (3.9%) seroreverted after a median follow‐up of 611 days. In a case‐control study, seroreversion was more likely to occur in patients with a higher weight (adjusted odds ratio aOR, 1.703; 95% confidence interval CI, 1.292‐2.323, per 10‐kg increment) and HIV viremia at the time of vaccination (aOR, 2.922; 95% CI, 1.067‐7.924), whereas positive seroresponse at 6 months of HAV vaccination and higher CD4 lymphocyte counts at vaccination were inversely associated with early seroreversion with an aOR of 0.059 (95% CI, 0.020‐0.154) and 0.837 (95% CI, 0.704‐0.979, per 100‐cell/mm3 increment), respectively, in multivariable analyses. Conclusion: During an outbreak setting, early seroreversion following two‐dose HAV vaccination occurred in 3.9% of HIV‐positive patients. Lower and delayed seroresponses to HAV vaccination, a higher weight, and HIV viremia and lower CD4 lymphocyte counts at the time of HAV vaccination were associated with early seroreversion. Regular monitoring of seroresponse and booster vaccination might be warranted, especially in HIV‐positive adults with predictors of early seroreversion.
Bio-inspired transverse brachiation robots mimic the movement of human climbers as they traverse along ledges on a vertical wall. The constraints on the locomotion of these robots differ considerably ...from those of conventional brachiation robots due primarily to the need for robust hand-eye coordination. This paper describes the development of a motion control strategy for a brachiation robot navigating between wall ledges positioned on a level plane or at different elevations. We based our robot on a four-link arm-body-tail system performing a four-phase movement, including a release phase, body reversal phase, swing-up phase, and grasping phase. We designed a gripper that uses passive wrist joint motion to grasp the ledge during the tail swing. We also developed a dynamic model by which to coordinate the swing-up movement, define the phase switching conditions, and time the grasping action of the grippers. In experiments, the robot proved highly effective in traversing between wall ledges of the same or different elevations.
Thioridazine (THD) is a common phenothiazine antipsychotic drug reported to suppress growth in several types of cancer cells. We previously showed that THD acts as an antiglioblastoma and anticancer ...stem-like cell agent. However, the signaling pathway underlying autophagy and apoptosis induction remains unclear. THD treatment significantly induced autophagy with upregulated AMPK activity and engendered cell death with increased sub-G1 in glioblastoma multiform (GBM) cell lines. Notably, through whole gene expression screening with THD treatment, frizzled (Fzd) proteins, a family of G-protein-coupled receptors, were found, suggesting the participation of Wnt/β-catenin signaling. After THD treatment, Fzd-1 and GSK3β-S9 phosphorylation (inactivated form) was reduced to promote β-catenin degradation, which attenuated P62 inhibition. The autophagy marker LC3-II markedly increased when P62 was released from β-catenin inhibition. Additionally, the P62-dependent caspase-8 activation that induced P53-independent apoptosis was confirmed by inhibiting T-cell factor/β-catenin and autophagy flux. Moreover, treatment with THD combined with temozolomide (TMZ) engendered increased LC3-II expression and caspase-3 activity, indicating promising drug synergism. In conclusion, THD induces autophagy in GBM cells by not only upregulating AMPK activity, but also enhancing P62-mediated autophagy and apoptosis through Wnt/β-catenin signaling. Therefore, THD is a potential alternative therapeutic agent for drug repositioning in GBM.
A 6.78-MHz wireless power receiver using a 3-mode reconfigurable resonant regulating rectifier for resonant wireless power transfer is presented. The proposed receiver improves power conversion ...efficiency and reduces die area and off-chip components by achieving power conversion plus voltage regulation in one stage, using only four on-chip power transistors and one off-chip capacitor. Moreover, the proposed 3-mode operation reduces the output voltage ripples and accomplishes switching synchronization easily during mode switching. The proposed pulsewidth modulation controller using ramp-stacking technique and type-II compensation achieves tight voltage regulation in the full loading range with fast transient responses. An adaptive sizing method is also employed to further improve the light-load efficiency of the receiver. Fabricated in a standard 0.35-μm CMOS process using 5-V devices, the receiver regulates the output voltage at 5 V and delivers a maximum power of 6 W. The measured peak efficiency reaches 92.2% when delivering an output power of 3.5 W. For a load step between 0.5 and 5 W, the overshoot and undershoot are less than 300 mV and the settling times are less than 16 μs.
Amphiphilic poly(hydroxyaminoethers) (PHAEs) with thermoresponsive solubility in aqueous solutions are developed in this study. Through the control of stoichiometric ratios of functional groups and ...the reaction temperature, a series of PHAEs with different hydrophilic segments is synthesized, as evidenced by proton nuclear magnetic resonance (1H‐NMR) and Fourier transform infrared (FT‐IR) spectroscopy. These PHAEs could be dissolved in aqueous solutions and exhibit the thermal phase transitions with lower critical solution temperatures (LCSTs) due to the hydrogen bonding between the hydrophilic segments and water, as confirmed by differential scanning calorimetry (DSC). In addition, the hydrophilic‐lipophilic balance (HLB) values suggest that the PHAEs are more hydrophilic amphiphiles and suitable for use as oil‐in‐water emulsifiers. Consequently, commercially available epoxy resins can be successfully stabilized by the PHAEs in water to form stable emulsions. These crosslinkable waterborne epoxy resins show good thermal and resistance properties, and the feasibility in anticorrosion coating applications for steel surfaces is demonstrated.
Amphiphilic poly(hydroxyaminoethers) exhibiting thermoresponsive solubility in aqueous solutions are developed, which are capable of emulsifying epoxy resins for anticorrosion coating applications.
Granular activated carbon (GAC) was impregnated with iron through a new multi-step procedure using ferrous chloride as the precursor for removing arsenic from drinking water. Scanning electron ...microscopy (SEM) and energy dispersive X-ray spectroscopy (EDS) analysis demonstrated that the impregnated iron was distributed evenly on the internal surface of the GAC. Impregnated iron formed nano-size particles, and existed in both crystalline (akaganeite) and amorphous iron forms. Iron-impregnated GACs (Fe-GACs) were treated with sodium hydroxide to stabilize iron in GAC and impregnated iron was found very stable at the common pH range in water treatments. Synthetic arsenate-contaminated drinking water was used in isotherm tests to evaluate arsenic adsorption capacities and iron use efficiencies of Fe-GACs with iron contents ranging from 1.64% to 12.13% (by weight). Nonlinear regression was used to obtain unbiased estimates of Langmuir model parameters. The arsenic adsorption capacity of Fe-GAC increased significantly with impregnated iron up to 4.22% and then decreased with more impregnated iron. Fe-GACs synthesized in this study exhibited higher affinity for arsenate as compared with references in literature and shows great potential for real implementations.
To investigate the effect of oroxylin A on the survival of retinal ganglion cells (RGC) and the activation of microglial cells in a rat optic nerve (ON) crush model.
Oroxylin A (15mg/Kg in 0.2ml ...phosphate-buffered saline) or phosphate-buffered saline (PBS control) was immediately administered after ON crush once by subcutaneous injection. Rats were euthanized at 2 weeks after the crush injury. The density of RGC was counted by retrograde labeling with FluoroGold and immunostaining of retina flat mounts for Brn3a. Electrophysiological visual function was assessed by flash visual evoked potentials (FVEP). TUNEL assay, immunoblotting analysis of glial fibrillary acidic protein (GFAP), inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in the retinas, and immunohistochemistry of GFAP in the retinas and ED1 in the ON were evaluated.
Two weeks after the insult, the oroxylin A-treated group had significantly higher FG labeled cells and Brn3a+ cells suggesting preserved RGC density in the central and mid-peripheral retinas compared with those of the PBS-treated group. FVEP measurements showed a significantly better preserved latency of the P1 wave in the ON-crushed, oroxylin A-treated rats than the ON-crushed, PBS treated rats. TUNEL assays showed fewer TUNEL positive cells in the ON-crushed, oroxylin A-treated rats. The number of ED1 positive cells was reduced at the lesion site of the optic nerve in the ON-crushed, oroxylin A-treated group. Increased GFAP expression in the retina was reduced greatly in ON-crushed, oroxylin A-treated group. Furthermore, administration of oroxylin A significantly attenuated ON crush insult-induced iNOS and COX-2 expression in the retinas.
These results demonstrated that oroxylin A hasss neuroprotective effects on RGC survival with preserved visual function and a decrease in microglial infiltration in the ONs after ON crush injury.
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