Synergistic production of TNF\u3b1 and IFN\u3b1 by human pDCs incubated with IFN\u3bb3 and IL-3

In this study, we investigated whether IFN\u3bb3 and IL-3 reciprocally influence their capacity to activate various functions of human plasmacytoid dendritic cells (pDCs). In fact, we preliminarily observed that IFN\u3bb3 upregulates the expression of the IL-3R\u3b1 (CD123), while IL-3 augments the expression of IFN\u3bbR1 in pDCs. As a result, we found that combination of IFN\u3bb3 and IL-3 induces a strong potentiation in the production of TNF\u3b1, IFN\u3b1, as well as in the expression of Interferon-Stimulated Gene (ISG) mRNAs by pDCs, as compared to either IFN\u3bb3 or IL-3 alone. In such regard, we found that endogenous IFN\u3b1 autocrinally promotes the expression of ISG mRNAs in IL-3-, but not in IFN\u3bb3 plus IL-3-, treated pDCs. Moreover, we uncovered that the production of IFN\u3b1 by IFN\u3bb3 plus IL-3-treated pDCs is mostly dependent on endogenously produced TNF\u3b1. Altogether, our data demonstrate that IFN\u3bb3 and IL-3 collaborate to promote, at maximal levels, discrete functional responses of human pDCs

Efficient Delivery of MicroRNA and AntimiRNA Molecules Using an Argininocalix[4]arene Macrocycle

MicroRNAs (miRNAs) are short non-coding RNA molecules acting as gene regulators by repressing translation or by inducing degradation of the target RNA transcripts. Altered expression of miRNAs may be involved in the pathogenesis of many severe human diseases, opening new avenues in the field of therapeutic strategies, i.e., miRNA targeting or miRNA mimicking. In this context, the efficient and non-toxic delivery of premiRNA and antimiRNA molecules might be of great interest. The aim of the present paper is to determine whether an argininocalix[4]arene is able to efficiently deliver miRNA, premiRNA, and antimiRNA molecules to target cells, preserving their biological activity. This study points out that (1) the toxicity of argininocalix[4]arene 1 is low, and it can be proposed for long-term treatment of target cells, being that this feature is a pre-requisite for the development of therapeutic protocols; (2) the delivery of premiRNA and antimiRNA molecules is efficient, being higher when compared with reference gold standards available; and (3) the biological activity of the premiRNAs and antimiRNAs is maintained. This was demonstrated using the argininocalix[4]arene 1 in miRNA therapeutic approaches performed on three well-described experimental model systems: (1) the induction of apoptosis by antimiR-221 in glioma U251 cells; (2) the induction of apoptosis by premiR-124 in U251 cells; and (3) the inhibition of pro-inflammatory IL-8 and IL-6 genes in cystic fibrosis IB3-1 cells. Our results demonstrate that the argininocalix[4]arene 1 should be considered a very useful delivery system for efficient transfer to target cells of both premiRNA and antimiRNA molecules, preserving their biological activity

Tuning the Loading and Release Properties of MicroRNA-Silencing Porous Silicon Nanoparticles by Using Chemically Diverse Peptide Nucleic Acid Payloads

Peptide nucleic acids (PNAs) are a class of artificial oligonucleotide mimics that have garnered much attention as precision biotherapeutics for their efficient hybridization properties and their exceptional biological and chemical stability. However, the poor cellular uptake of PNA is a limiting factor to its more extensive use in biomedicine; encapsulation in nanoparticle carriers has therefore emerged as a strategy for internalization and delivery of PNA in cells. In this study, we demonstrate that PNA can be readily loaded into porous silicon nanoparticles (pSiNPs) following a simple salt-based trapping procedure thus far employed only for negatively charged synthetic oligonucleotides. We show that the ease and versatility of PNA chemistry also allows for producing PNAs with different net charge, from positive to negative, and that the use of differently charged PNAs enables optimization of loading into pSiNPs. Differently charged PNA payloads determine different release kinetics and allow modulation of the temporal profile of the delivery process. In vitro silencing of a set of specific microRNAs using a pSiNP-PNA delivery platform demonstrates the potential for biomedical applications

Child Neurology: A Case Series of Heterogeneous Neuropsychiatric Symptoms and Outcome in Very Early-Onset Narcolepsy Type 1

Narcolepsy type 1 is a central disorder of hypersomnolence characterized by excessive daytime sleepiness, cataplexy (i.e., sudden loss of muscle tone during wakefulness triggered by emotions), and REM sleep-related manifestations that can present with a peculiar phenotype when arising at a pediatric age. Several features of childhood-onset narcolepsy type 1 are also common in neuropsychiatric conditions; discrete neuropsychiatric comorbidity has also been demonstrated. Here, we report on 3 children with very early narcolepsy type 1. All 3 patients had psychiatric features at the time of symptom onset coupled with peculiar motor disturbances. The course of narcolepsy symptoms also paralleled neuropsychiatric symptoms, suggesting a possible intrinsic link between sleep and psychological features. Multidisciplinary management is mandatory for pediatric narcolepsy type 1 since prompt disease management addressing neuropsychiatric symptoms could lead to better clinical outcomes and quality of life

An active feedback recovery technique from disruption events induced by m=2 n=1 tearing modes in ohmically heated tokamak plasmas

We present experimental results of magnetic feedback control on the m=2, n=1 tearing mode in RFX-mod operated as a circular ohmically heated tokamak. The feedback suppression of the non-resonant m=2, n=1 Resistive Wall Mode (RWM) in q(a)<2 plasmas is a well-established result of RFX-mod. The control of the tearing counterpart, which develops in q(a)>2 equilibrium, is instead a more difficult issue. In fact, the disruption induced by a growing amplitude m=2, n=1 tearing mode can be prevented by feedback only when the resonant surface q=2 is close to the plasma edge, namely 2<q(a)<2.5, and the electron density does not exceed approximately half of the Greenwald limit. A combined technique of tearing mode and q(a) control has been therefore developed to recover the discharge from the most critical conditions: the potentially disruptive tearing mode is converted into the relatively benign RWM by suddenly decreasing q(a) below 2. The experiments demonstrate the concept with 100% of successful cases. The q(a) control has been performed through the plasma current, given the capability of the toroidal loop-voltage power supply of RFX-mod. We also propose a path for controlling q(a) by acting on the plasma shape, which could be applied to medium size elongated tokamaks

A peptide-nucleic acid targeting miR-335-5p enhances expression of cystic fibrosis transmembrane conductance regulator (CFTR) gene with the possible involvement of the CFTR scaffolding protein NHERF1

(1) Background: Up-regulation of the Cystic Fibrosis Transmembrane Conductance Regulator gene (CFTR) might be of great relevance for the development of therapeutic protocols for cystic fibrosis (CF). MicroRNAs are deeply involved in the regulation of CFTR and scaffolding proteins (such as NHERF1, NHERF2 and Ezrin). (2) Methods: Content of miRNAs and mRNAs was analyzed by RT-qPCR, while the CFTR and NHERF1 production was analyzed by Western blotting. (3) Results: The results here described show that the CFTR scaffolding protein NHERF1 can be upregulated in bronchial epithelial Calu-3 cells by a peptide-nucleic acid (PNA) targeting miR-335-5p, predicted to bind to the 3′-UTR sequence of the NHERF1 mRNA. Treatment of Calu-3 cells with this PNA (R8-PNA-a335) causes also up-regulation of CFTR. (4) Conclusions: We propose miR-335-5p targeting as a strategy to increase CFTR. While the efficiency of PNA-based targeting of miR-3355p should be verified as a therapeutic strategy in CF caused by stop-codon mutation of the CFTR gene, this approach might give appreciable results in CF cells carrying other mutations impairing the processing or stability of CFTR protein, supporting its application in personalized therapy for precision medicine

Corilagin Induces High Levels of Apoptosis in the Temozolomide-Resistant T98G Glioma Cell Line

Glioblastoma multiforme (GBM), a malignant tumor of the central nervous system, has a high mortality rate; no curativetreatment is presently available and the most commonly used chemiotherapeutic drug, the alkylating agent temozolomide (TMZ), is only able to increase life expectancy and is often associated with drugresistance. Therefore, an urgent need does exist for novel drugs aimed at treating gliomas. In the present study we obtained three major results using corliagin: (a) demonstrate that it inhibits the growth of U251 glioma cells through activation of the apoptotic pathway; (b) demonstrate that it is also active on temozolomideresistant T98G glioma cells; (c) demonstrate that when used in combination with temozolomide on T98G glioma cells a higher level of pro-apototic and antiproliferative effects are observed. Our study indicates that corilagin should be investigated in more detail in order to determine if it can be developed as a potential therapeutic agent. In addition, our results suggest that corilagin could be used in combination with low dosages of other standard anticancer chemotherapeutic drugs against gliomas (such as temozolomide) with the aim of obtaining enhanced anticancer effects

Enzymatic spermine metabolites induce apoptosis associated with increase of p53, caspase-3 and mir-34a in both neuroblastoma cells, SJNKP and the N-Myc-amplified form IMR5

Neuroblastoma (NB) is a common malignant solid tumor in children and accounts for 15% of childhood cancer mortality. Amplification of the N-Myc oncogene is a well-established poor prognostic marker in NB patients and strongly correlates with higher tumor aggression and resistance to treatment. New therapies for patients with N-Myc-amplified NB need to be developed. After treating NB cells with BSAO/SPM, the detection of apoptosis was determined after annexin V-FITC labeling and DNA staining with propidium iodide. The mitochondrial membrane potential activity was checked, labeling cells with the probe JC-1 dye. We analyzed, by real-time RT-PCR, the transcript of genes involved in the apoptotic process, to determine possible down-or upregulation of mRNAs after the treatment on SJNKP and the N-Myc-amplified IMR5 cell lines with BSAO/SPM. The experiments were carried out considering the proapoptotic genes Tp53 and caspase-3. After treatment with BSAO/SPM, both cell lines displayed increased mRNA levels for all these proapoptotic genes. Western blotting analysis with PARP and caspase-3 antibody support that BSAO/SPM treatment induces high levels of apoptosis in cells. The major conclusion is that BSAO/SPM treatment leads to antiproliferative and cytotoxic activity of both NB cell lines, associated with activation of apoptosis