Effect of all-trans retinoic acid on the proliferation and differentiation of brain tumor stem cells

<p>Abstract</p> <p>Objective</p> <p>To investigate the effect of all-trans retinoic acid(ATRA) on the proliferation and differentiation of brain tumor stem cells(BTSCs) <it>in vitro</it>.</p> <p>Methods</p> <p>Limiting dilution and clonogenic assay were used to isolate and screen BTSCs from the fresh specimen of human brain glioblastoma. The obtained BTSCs, which were cultured in serum-free medium, were classified into four groups in accordance with the composition of the different treatments. The proliferation of the BTSCs was evaluated by MTT assay. The BTSCs were induced to differentiate in serum-containing medium, and classified into the ATRA group and control group. On the 10^thday of induction, the expressions of CD133 and glial fibrillary acidic protein (GFAP) in the differentiated BTSCs were detected by immunofluorescence. The differentiated BTSCs were cultured in serum-free medium, the percentage and the time required for formation of brain tumor spheres (BTS) were observed.</p> <p>Results</p> <p><b>BTSCs </b>obtained by limiting dilution were all identified as CD133-positive by immunofluorescence. In serum-free medium, the proliferation of BTSCs in the ATRA group was observed significantly faster than that in the control group, but slower than that in the growth factor group and ATRA/growth factor group, and the size of the BTS in the ATRA group was smaller than that in the latter two groups(<it>P </it>< 0.01). In serum-containing medium, the expression percentages of CD133 and GFAP in the differentiated BTSCs were (2.29% ± 0.27%) and (75.60% ± 4.03%) respectively in the ATRA group, and (7.05% ± 0.49%) and (12.51% ± 0.77%) respectively in the control group. The differentiation rate of BTSCs in the ATRA group was significantly higher than that in the control group (<it>P </it>< 0.05), but there was still CD133 expressed in the ATRA group. The differentiated BTSCs could re-form BTSs in serum-free medium. The percentage of BTS formation in the ATRA group was(4.84% ± 0.32%), significantly lower than that in the control group (17.71% ± 0.78%) (<it>P </it>< 0.05), and the time required for BTS formation in the ATRA group was (10.07 ± 1.03)d, significantly longer than that in the control group (4.08 ± 0.35)d (<it>P </it>< 0.05).</p> <p>Conclusion</p> <p><b>ATRA </b>can promote the proliferation and induce the differentiation of BTSCs, but the differentiation is incomplete, terminal differentiation cannot be achieved and BTSs can be formed again.</p

Production of proton-rich nuclei around Z=84-90 in fusion-evaporation reactions

Within the framework of the dinuclear system model, production cross sections of proton-rich nuclei with charged numbers of Z=84-90 are investigated systematically. Possible combinations with the $^{28}$Si, $^{32}$S, $^{40}$Ar bombarding the target nuclides $^{165}$Ho, $^{169}$Tm, $^{170-174}$Yb, $^{175,176}$Lu, $^{174,176-180}$Hf and $^{181}$Ta are analyzed thoroughly. The optimal excitation energies and evaporation channels are proposed to produce the proton-rich nuclei. The systems are feasible to be constructed in experiments. It is found that the neutron shell closure of N=126 is of importance during the evaporation of neutrons. The experimental excitation functions in the $^{40}$Ar induced reactions can be nicely reproduced. The charged particle evaporation is comparable with neutrons in cooling the excited proton-rich nuclei, in particular for the channels with $\alpha$ and proton evaporation. The production cross section increases with the mass asymmetry of colliding systems because of the decrease of the inner fusion barrier. The channels with pure neutron evaporation depend on the isotopic targets. But it is different for the channels with charged particles and more sensitive to the odd-even effect.Comment: 15 pages, 10 figures. arXiv admin note: text overlap with arXiv:0803.1117, arXiv:0707.258

N6-methyl-adenosine (m6A) in RNA: An Old Modification with A Novel Epigenetic Function

AbstractN6-methyl-adenosine (m6A) is one of the most common and abundant modifications on RNA molecules present in eukaryotes. However, the biological significance of m6A methylation remains largely unknown. Several independent lines of evidence suggest that the dynamic regulation of m6A may have a profound impact on gene expression regulation. The m6A modification is catalyzed by an unidentified methyltransferase complex containing at least one subunit methyltransferase like 3 (METTL3). m6A modification on messenger RNAs (mRNAs) mainly occurs in the exonic regions and 3′-untranslated region (3′-UTR) as revealed by high-throughput m6A-seq. One significant advance in m6A research is the recent discovery of the first two m6A RNA demethylases fat mass and obesity-associated (FTO) gene and ALKBH5, which catalyze m6A demethylation in an α-ketoglutarate (α-KG)- and Fe2+-dependent manner. Recent studies in model organisms demonstrate that METTL3, FTO and ALKBH5 play important roles in many biological processes, ranging from development and metabolism to fertility. Moreover, perturbation of activities of these enzymes leads to the disturbed expression of thousands of genes at the cellular level, implicating a regulatory role of m6A in RNA metabolism. Given the vital roles of DNA and histone methylations in epigenetic regulation of basic life processes in mammals, the dynamic and reversible chemical m6A modification on RNA may also serve as a novel epigenetic marker of profound biological significances

Activation of mammalian target of rapamycin mediates rat pain-related responses induced by BmK I, a sodium channel-specific modulator

The mammalian target of rapamycin (mTOR) is known to regulate cell proliferation and growth by controlling protein translation. Recently, it has been shown that mTOR signaling pathway is involved in long-term synaptic plasticity. However, the role of mTOR under different pain conditions is less clear. In this study, the spatiotemporal activation of mTOR that contributes to pain-related behaviors was investigated using a novel animal inflammatory pain model induced by BmK I, a sodium channel-specific modulator purified from scorpion venom. In this study, intraplantar injections of BmK I were found to induce the activation of mTOR, p70 ribosomal S6 protein kinase (p70 S6K) and eukaryotic initiation factor 4E-binding protein 1 (4E-BP1) in rat L5-L6 spinal neurons. In the spinal cord, mTOR, p70 S6K and 4E-BP1 were observed to be activated in the ipsilateral and contralateral regions, peaking at 1-2 h and recovery at 24 h post-intraplantar (i.pl.) BmK I administration. In addition, intrathecal (i.t.) injection of rapamycin - a specific inhibitor of mTOR - was observed to result in the reduction of spontaneous pain responses and the attenuation of unilateral thermal and bilateral mechanical hypersensitivity elicited by BmK I. Thus, these results indicate that the mTOR signaling pathway is mobilized in the induction and maintenance of pain-activated hypersensitivity

Evaluation of parameters affecting Agrobacterium-mediated transient expression in citrus

Abstract Agrobacterium-mediated transient expression assays are a convenient alternative to stable expression because they are simple, easy to perform, and achieve gene expression rapidly. This study investigated the factors affecting transient gene expression efficiency in citrus by observing the cryo-sectioning of leaf samples under a laser confocal microscope. These factors included the composition of the infiltration buffer, the Agrobacterium cell density, the leaf development stage, the incubation temperature, and plant genotype. The highest transient expression level of yellow fluorescent protein (YFP) was detected in Mexican lime (Citrus aurantifolia) on the third day after the intermediate-aged leaves were infiltrated with the improved infiltration buffer 1 (15 mmol L−1 2-(N-morpholino) ethanesulfonic acid, 10 mmol L−1 MgCI2, and 200 pmol L−1 acetosyringone), which had an optical density of 0.8 and was incubated at 22°C. Additionally, this transient expression assay was applied to other citrus genotypes. Of note, trifoliate orange (Poncirus trifoliata) and kumquat (Fortunella obovate) had higher expression efficiency than other six genotypes of the Citrus genus. Our study provides research basis for the selection of optimization strategies in transient gene expression and improves the method for available genome investigation in citrus

Zero-Bias Deep Learning for Accurate Identification of Internet of Things (IoT) Devices

The Internet of Things (IoT) provides applications and services that would otherwise not be possible. However, the open nature of IoT makes it vulnerable to cybersecurity threats. Especially, identity spoofing attacks, where an adversary passively listens to the existing radio communications and then mimic the identity of legitimate devices to conduct malicious activities. Existing solutions employ cryptographic signatures to verify the trustworthiness of received information. In prevalent IoT, secret keys for cryptography can potentially be disclosed and disable the verification mechanism. Noncryptographic device verification is needed to ensure trustworthy IoT. In this article, we propose an enhanced deep learning framework for IoT device identification using physical-layer signals. Specifically, we enable our framework to report unseen IoT devices and introduce the zero-bias layer to deep neural networks to increase robustness and interpretability. We have evaluated the effectiveness of the proposed framework using real data from automatic dependent surveillance-broadcast (ADS-B), an application of IoT in aviation. The proposed framework has the potential to be applied to the accurate identification of IoT devices in a variety of IoT applications and services