Direct transformation of dinitrogen: synthesis of N-containing organic compounds via N−C bond formation

N-containing organic compounds are of vital importance to lives. Practical synthesis of valuable N-containing organic compounds directly from dinitrogen (N₂), not through ammonia (NH₃), is a holy-grail in chemistry and chemical industry. An essential step for this transformation is the functionalization of the activated N₂ units/ligands to generate N−C bonds. Pioneering works of transition metal-mediated direct conversion of N₂ into organic compounds via N−C bond formation at metal-dinitrogen [N₂-M] complexes have generated diversified coordination modes and laid the foundation of understanding for the N−C bond formation mechanism. This review summarizes those major achievements and is organized by the coordination modes of the [N₂-M] complexes (end-on, side-on, end-on-side-on, etc.) that are involved in the N−C bond formation steps, and each part is arranged in terms of reaction types (N-alkylation, N-acylation, cycloaddition, insertion, etc.) between [N₂-M] complexes and carbon-based substrates. Additionally, earlier works on one-pot synthesis of organic compounds from N₂ via ill-defined intermediates are also briefed. Although almost all of the syntheses of N-containing organic compounds via direct transformation of N₂ so far in the literature are realized in homogeneous stoichiometric thermochemical reaction systems and are discussed here in detail, the sporadically reported syntheses involving photochemical, electrochemical, heterogeneous thermo-catalytic reactions, if any, are also mentioned. This review aims to provide readers with an in-depth understanding of the state-of-the-art and perspectives of future research particularly in direct catalytic and efficient conversion of N₂ into N-containing organic compounds under mild conditions, and to stimulate more research efforts to tackle this long-standing and grand scientific challenge.     

Disruption of splicing-regulatory elements using CRISPR/Cas9 to rescue spinal muscular atrophy in human iPSCs and mice

We here report a genome-editing strategy to correct spinal muscular atrophy (SMA). Rather than directly targeting the pathogenic exonic mutations, our strategy employed Cas9 and guide-sgRNA for the targeted disruption of intronic splicing-regulatory elements. We disrupted intronic splicing silencers (ISSs, including ISS-N1 and ISS + 100) of survival motor neuron (SMN) 2, a key modifier gene of SMA, to enhance exon 7 inclusion and full-length SMN expression in SMA iPSCs. Survival of splicing-corrected iPSC-derived motor neurons was rescued with SMN restoration. Furthermore, co-injection of Cas9 mRNA from Streptococcus pyogenes (SpCas9) or Cas9 from Staphylococcus aureus (SaCas9) alongside their corresponding sgRNAs targeting ISS-N1 into zygotes rescued 56% and 100% of severe SMA transgenic mice (Smn^−/−, SMN2^tg/−). The median survival of the resulting mice was extended to >400 days. Collectively, our study provides proof-of-principle for a new strategy to therapeutically intervene in SMA and other RNA-splicing-related diseases.     

Multiple-prespecified-dictionary sparse representation for compressive sensing image reconstruction with nonconvex regularization

Multiple-prespecified-dictionary sparse representation (MSR) has shown powerful potential in compressive sensing (CS) image reconstruction, which can exploit more sparse structure and prior knowledge of images for minimization. Due to the popular L₁ regularization can only achieve the suboptimal solution of L₀ regularization, using the nonconvex regularization can often obtain better results in CS reconstruction. This paper proposes a nonconvex adaptive weighted L_p regularization CS framework via MSR strategy. We first proposed a nonconvex MSR based L_p regularization model, then we propose two algorithms for minimizing the resulting nonconvex L_p optimization problem. According to the fact that the sparsity levels of each regularizers are varying with these prespecified-dictionaries, an adaptive scheme is proposed to weight each regularizer for optimization by exploiting the difference of sparsity levels as prior knowledge. Simulated results show that the proposed nonconvex framework can make a significant improvement in CS reconstruction than convex L₁ regularization, and the proposed MSR strategy can also outperforms the traditional nonconvex L_p regularization methodology.     

Diet high in branched-chain amino acid promotes PDAC development by USP1-mediated BCAT2 stabilization

BCAT2-mediated branched-chain amino acid (BCAA) catabolism is critical for pancreatic ductal adenocarcinoma (PDAC) development, especially at an early stage. However, whether a high-BCAA diet promotes PDAC development in vivo, and the underlying mechanism of BCAT2 upregulation, remain undefined. Here, we find that a high-BCAA diet promotes pancreatic intraepithelial neoplasia (PanIN) progression in LSL-Kras^G12D/+; Pdx1-Cre (KC) mice. Moreover, we screened with an available deubiquitylase library which contains 31 members of USP family and identified that USP1 deubiquitylates BCAT2 at the K229 site. Furthermore, BCAA increases USP1 protein at the translational level via the GCN2-eIF2α pathway both in vitro and in vivo. More importantly, USP1 inhibition recedes cell proliferation and clone formation in PDAC cells and attenuates pancreas tumor growth in an orthotopic transplanted mice model. Consistently, a positive correlation between USP1 and BCAT2 is found in KC; LSL-Kras^G12D/+; p53^flox/+; Pdx1-Cre mice and clinical samples. Thus, a therapeutic targeting USP1-BCAT2-BCAA metabolic axis could be considered as a rational strategy for treatment of PDAC and precisive dietary intervention of BCAA has potentially translational significance.     

Mixed H∞ and passive control for a class of nonlinear switched systems with average dwell time via hybrid control approach

This paper investigates the mixed H_∞ and passive control problem for a class of nonlinear switched systems based on a hybrid control strategy. To solve this problem, firstly, using the Takagi–Sugeno (T–S) fuzzy model to approximate every nonlinear subsystem, the nonlinear switched systems are modeled as the switched T–S fuzzy systems. Secondly, the hybrid controllers are used to stabilize the switched T–S fuzzy systems. The hybrid controllers consist of dynamic output-feedback controllers for every subsystem and state updating controllers at the switching instant. Thirdly, a new performance index is proposed for switched systems. This new performance index can be viewed as the mixed weighted H_∞ and passivity performance. Based on this new performance index, the weighted H_∞ control problem and the passive control problem for switched T–S fuzzy systems via the hybrid control strategy are solved in a unified framework. Together the multiple Lyapunov functions (MLFs) approach with the average dwell time (ADT) technique, new design conditions for the hybrid controllers are obtained. Under these conditions, the closed-loop switched T–S fuzzy systems are globally uniformly asymptotically stable with a prescribed mixed H_∞ and passivity performance index. Moreover, the desired hybrid controllers can be constructed by solving a set of linear matrix inequalities (LMIs). Finally, the effectiveness of the obtained results is illustrated by a numerical example.     

Inhibitory and bactericidal activity of the calmodulin antagonist,trifluoperazine, againstMycobacterium avium in vitro and within human monocyte-derived macrophages

The activity of a calmodulin antagonist, trifluoperazine (TFP), was testedin vitro againstMycobacterium avium (ATCC 25291). The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of this compound forM. avium were 20 and 30 μg/ml, respectively. TFP was also found to completely inhibit the growth of 10 isolates ofM. avium (5 patient isolates and 5 environmental isolates) at 30 μg/ml. At near neutral pH (6.8), the MIC of TFP was found to be 20 μg/ml. However, at pH 5.5 (intracellular pH of macrophages), there was a decrease in the inhibitory activity of the compound against this organism. Interestingly, 99.6% ofM. avium within human monocyte-derived macrophages were killed at a drug concentration of 30 μg/ml, which correlates well with the MBC of TFP againstM. avium in vitro. Although the MIC for TFP appears to be higher than that forMycobacterium tuberculosis H₃₇R_v, our studies suggest that calmodulin antagonists might be useful as drugs against infection due toM. avium. It is suggested that administration of TFP in combination with other known drugs may enhance the overall bactericidal effect.     

Rapid magnetic heating treatment by highly charged maghemite nanoparticles on Wistar rats exocranial glioma tumors at microliter volume

One of the most significant challenges implementing colloidal magnetic nanoparticles in medicine is the efficient heating of microliter quantities by applying a low frequency alternating magnetic field. The ultimate goal is to accomplish nonsurgically the treatment of millimeter size tumors. Here, we demonstrate the synthesis, characterization, and the in vitro as well as in vivo efficiency of a dextran coated maghemite (γ-Fe₂O₃) ferrofluid with an exceptional response to magnetic heating. The difference to previous synthetic attempts is the high charge of the dextran coating, which according to our study maintains the colloidal stability and good dispersion of the ferrofluid during the magnetic heating stage. Specifically, in vitro 2 μl of the ferrofluid gives an outstanding temperature rise of 33 °C within 10 min, while in vivo treatment, by infusing 150 μl of the ferrofluid in animal model (rat) glioma tumors, causes an impressive cancer tissue dissolution.     

Periodic solutions to systems of reaction-diffusion equations

In this paper, necessary and sufficient conditions are derived for the existence of temporally periodic “dissipative structure” solutions in cases of weak diffusion with the reaction rate terms dominant in a generic system of reaction-diffusion equations ?c_i/?t = D_i?²c_i+Q_i(c), where the enumerator index i runs 1 to n, c_i = c_i(x, t) denotes the concentration or density of the ith participating molecular or biological species, D_i is the diffusivity constant for the ith species and Q_i(c), an algebraic function of the n-tuple c = (c₁,\3., c_n), expresses the local rate of production of the ith species due to chemical reactions or biological interactions.     

Au23(CR)14 nanocluster restores fibril Aβ’s unfolded state with abolished cytotoxicity and dissolves endogenous Aβ plaques

The misfolding of amyloid-β (Aβ) peptides from the natural unfolded state to β-sheet structure is a critical step, leading to abnormal fibrillation and formation of endogenous Aβ plaques in Alzheimer''s disease (AD). Previous studies have reported inhibition of Aβ fibrillation or disassembly of exogenous Aβ fibrils in vitro. However, soluble Aβ oligomers have been reported with increased cytotoxicity; this might partly explain why current clinical trials targeting disassembly of Aβ fibrils by anti-Aβ antibodies have failed so far. Here we show that Au₂₃(CR)₁₄ (a new Au nanocluster modified by Cys-Arg (CR) dipeptide) is able to completely dissolve exogenous mature Aβ fibrils into monomers and restore the natural unfolded state of Aβ peptides from misfolded β-sheets. Furthermore, the cytotoxicity of Aβ₄₀ fibrils when dissolved by Au₂₃(CR)₁₄ is fully abolished. More importantly, Au₂₃(CR)₁₄ is able to completely dissolve endogenous Aβ plaques in brain slices from transgenic AD model mice. In addition, Au₂₃(CR)₁₄ has good biocompatibility and infiltration ability across the blood–brain barrier. Taken together, this work presents a promising therapeutics candidate for AD treatment, and manifests the potential of nanotechnological approaches in the development of nanomedicines.     

Synthesis of silver nanoparticles using a biosurfactant produced in low-cost medium as stabilizing agent

BackgroundA biosurfactant produced by Pseudomonas aeruginosa cultivated in a low-cost medium formulated with 2.5% vegetable oil refinery residue and 2.5% corn steep liquor and distilled water was employed to stabilize silver nanoparticles in the liquid phase. The particles were initially synthesized using NaBH₄ as reducing agent in biosurfactant reverse micelles and were extracted from the micellar solution to disperse in heptane.ResultsA silver particle size in the range of 1.13 nm was observed. The UV–vis absorption spectra proposed that silver nanoparticles could be formed in the reverse micelles and relatively stabilized for at least 3 months without passivator addition. The Transmission Electron Microscope (TEM) shows that the silver nanoparticles are of spherical form and relatively uniform.ConclusionsThis process provided a simpler route for nanoparticle synthesis compared to existing systems using whole organisms or partially purified biological extracts, showing that the low-cost biosurfactant can be used for nanoparticle synthesis as a non-toxic and biodegradable stabilizing agent.     

Modeling Arabic subjectivity and sentiment in lexical space

In spite of the vast amount of work on subjectivity and sentiment analysis (SSA), it is not yet particularly clear how lexical information can best be modeled in a morphologically-richness language. To bridge this gap, we report successful models targeting lexical input in Arabic, a language of very complex morphology. Namely, we measure the impact of both gold and automatic segmentation on the task and build effective models achieving significantly higher than our baselines. Our models exploiting predicted segments improve subjectivity classification by 6.02% F₁-measure and sentiment classification by 4.50% F₁-measure against the majority class baseline surface word forms. We also perform in-depth (error) analyses of the behavior of the models and provide detailed explanations of subjectivity and sentiment expression in Arabic against the morphological richness background in which the work is situated.     

Effects of shear stresses and antioxidant concentrations on the production of reactive oxygen species in lung cancer cells

Reactive oxygen species (ROS) are known to be a key factor in the development of cancer, and many exogenous sources are supposed to be related to the formation of ROS. In this paper, a microfluidic chip was developed for studying the production of ROS in lung cancer cells under different chemical and physical stimuli. This chip has two unique features: (1) five relative concentrations of 0, 1/8, 1/2, 7/8, and 1 are achieved in the culture regions; (2) a shear stress gradient is produced inside each of the five culture areas. Lung cancer cells were seeded inside this biocompatible chip for investigating their response to different concentrations of H₂O₂, a chemical stimulus known to increase the production of ROS. Then the effect of shear stress, a physical stimulus, on lung cancer cells was examined, showing that the production of ROS was increased in response to a larger shear stress. Finally, two antioxidants, α-tocopherol and ferulic acid, were used to study their effects on reducing ROS. It was found that high-dose α-tocopherol was not able to effectively eliminate the ROS produced inside cells. This counter effect was not observed in cells cultured in a traditional chamber slide, where no shear stress was present. This result suggests that the current microfluidic chip provides an in vitro platform best mimicking the physiological condition where cells are under circulating conditions.     

Improvement of anther cultures conditions using the Taguchi method in three cereal crops

BackgroundPlant tissue cultures have the potential to reprogram the development of microspores from normal gametophytic to sporophytic pathway resulting in the formation of androgenic embryos. The efficiency of this process depends on the genotype, media composition and external conditions. However, this process frequently results in the regeneration of albino instead of green plants. Successful regeneration of green plants is affected by the concentration of copper sulfate (CuSO₄) and silver nitrate (AgNO₃) and the length of induction step. In this study, we aimed at concurrent optimization of these three factors in barley (Hordeum vulgare L.), wheat (Triticum aestivum L.), and triticale (x Triticosecale spp. Wittmack ex A. Camus 1927) using the Taguchi method. We evaluated uniform donor plants under varying experimental conditions of in vitro anther culture using the Taguchi approach, and verified the optimized conditions.ResultsOptimization of the regeneration conditions resulted in an increase in the number of green regenerants compared with the control. Statistic Taguchi method for optimization of the in vitro tissue culture plant regeneration via anther cultures allowed reduction of the number of experimental designs from 27 needed if full factorial analysis is used to 9. With the increase in the number of green regenerants, the number of spontaneous doubled haploids decreased. Moreover, in barley and triticale, the number of albino regenerants was reduced.ConclusionThe statistic Taguchi approach could be successfully used for various factors (here components of induction media, time of incubation on induction media) at a one time, that may impact on cereals anther cultures to improve the regeneration efficiency.How to cite: Orłowska R, Pachota KA, Machczyńska J, et al. Improvement of anther cultures conditions using the Taguchi method in three cereal crops. Electron J Biotechnol 2020;43. https://doi.org/10.1016/j.ejbt.2019.11.001.     

An in vitro model mimicking the complement system to favor directed phagocytosis of unwanted cells

BackgroundOpsonization, is the molecular mechanism by which target molecules promote interactions with phagocyte cell surface receptors to remove unwanted cells by induced phagocytosis. We designed an in vitro system to demonstrate that this procedure could be driven to eliminate adipocytes, using peptides mimicking regions of the complement protein C3b to promote opsonization and enhance phagocytosis. Two cell lines were used: (1) THP-1 monocytes differentiated to macrophages, expressing the C3b opsonin receptor CR1 in charge of the removal of unwanted coated complexes; (2) 3T3-L1 fibroblasts differentiated to adipocytes, expressing AQP7, to evaluate the potential of peptides to stimulate opsonization. (3) A co-culture of the two cell lines to demonstrate that phagocytosis could be driven to cell withdrawal with high efficiency and specificity.ResultsAn array of peptides were designed and chemically synthesized p3691 and p3931 joined bound to the CR1 receptor activating phagocytosis (p < 0.033) while p3727 joined the AQP7 protein (p < 0.001) suggesting that opsonization of adipocytes could occur. In the co-culture system p3980 and p3981 increased lipid uptake to 91.2% and 89.0%, respectively, as an indicator of potential adipocyte phagocytosis.ConclusionsThis in vitro model could help understand the receptor–ligand interaction in the withdrawal of unwanted macromolecules in vivo. The adipocyte-phagocytosis discussed may help to control obesity, since peptides of C3b stimulated the CR1 receptor, promoting opsonisation and phagocytosis of lipid-containing structures, and recognition of AQP7 in the differentiated adipocytes, favored the phagocytic activity of macrophages, robustly supported by the co-culture strategy.How to cite: Bartsch IM, Perelmuter K, Bollati-Fogolin M, et al. An in vitro model mimicking the complement system to favor directed phagocytosis of unwanted cells. Electron J Biotechnol 2021;49. https://doi.org/10.1016/j.ejbt.2020.09.010.     

Stability, L1-gain analysis and asynchronous L1-gain control of uncertain discrete-time switched positive linear systems with dwell time

In this paper, the stability, L₁-gain analysis and asynchronous L₁-gain control problems of uncertain discrete-time switched positive linear systems (DSPLSs) with dwell time are investigated. First, several convex and non-convex conditions on dwell time stability of DSPLSs with interval and polytopic uncertainties are presented, and the relation between these conditions is revealed. Then, via a switched dwell-time-dependent co-positive Lyapunov functions (SDTLFs) approach, convex sufficient conditions on L₁-gain analysis and asynchronous L₁-gain control of DSPLSs with interval uncertainties are derived. Meanwhile, via the switched parameter-dwell-time-dependent co-positive Lyapunov functions (SPDTLFs) approach, the L₁-gain analysis and asynchronous L₁-gain controller design problems of DSPLSs with polytopic uncertainties are also solved. The stability and L₁-gain analysis results are given in terms of linear programming (LP). The controller design results are presented in terms of bilinear programming (BP), which can be solved with the help of iterative algorithm. At last, both numerical and practical examples are provided to show the effectiveness of the results.     

Spheroid-on-chip microfluidic technology for the evaluation of the impact of continuous flow on metastatic potential in cancer models in vitro

The majority of cancer deaths are linked to tumor spread, or metastasis, but 3D in vitro metastasis models relevant to the tumor microenvironment (including interstitial fluid flow) remain an area of unmet need. Microfluidics allows us to introduce controlled flow to an in vitro cancer model to better understand the relationship between flow and metastasis. Here, we report new hybrid spheroid-on-chip in vitro models for the impact of interstitial fluid flow on cancer spread. We designed a series of reusable glass microfluidic devices to contain one spheroid in a microwell under continuous perfusion culture. Spheroids derived from established cancer cell lines were perfused with complete media at a flow rate relevant to tumor interstitial fluid flow. Spheroid viability and migratory/invasive capabilities were maintained on-chip when compared to off-chip static conditions. Importantly, using flow conditions modeled in vitro, we are the first to report flow-induced secretion of pro-metastatic factors, in this case cytokines vascular endothelial growth factor and interleukin 6. In summary, we have developed a new, streamlined spheroid-on-chip in vitro model that represents a feasible in vitro alternative to conventional murine in vivo metastasis assays, including complex tumor environmental factors, such as interstitial fluid flow, extracellular matrices, and using 3D models to model nutrient and oxygen gradients. Our device, therefore, constitutes a robust alternative to in vivo early-metastasis models for determination of novel metastasis biomarkers as well as evaluation of therapeutically relevant molecular targets not possible in in vivo murine models.     

Expression,purification and biological effect of a novel single chain Fv antibody and protamine fusion protein for the targeted delivery of siRNAs to FGFR3 positive cancer cells

BackgroundGain-of-function of fibroblast growth factor receptor 3 (FGFR3) is involved in the pathogenesis of many tumors. More and more studies have focused on the potential usage of therapeutic single-chain Fv (ScFv) antibodies against FGFR3. RNA interference (RNAi) has been considered as a promising therapeutic method against cancer. A tool which can deliver small interference RNAs (siRNAs) into FGFR3 positive cancer cells is very promising for anti-tumor therapy.ResultsIn this study, a novel fusion protein R3P, which consists of FGFR3-ScFv and protamine, was generated in Escherichia coli by inclusion body expression strategy and Ni-NTA chromatography. Its yield reached 10 mg per liter of bacterial culture and its purity was shown to be higher than 95%. 1 μg of R3P could efficiently bind to about 2.5 pmol siRNAs and deliver siRNAs into FGFR3 positive RT112 and K562 cells. Annexin V staining results showed that R3P can deliver the amplified breast cancer 1 (AIB1) siRNAs to induce RT112 cell apoptosis.ConclusionThese results indicated that R3P was a promising carrier tool to deliver siRNAs into FGFR3 positive cancer cells and to exert anti-tumor effect.     

A green,efficient and precise hydrogen therapy of cancer based on in vivo electrochemistry

By combined use of traditional Chinese acupuncture Fe needle electrode and in vivo electrochemistry, we achieved in vivo H₂ generation in tumors in a controllable manner and exploited it for effective and green therapy of tumors for the first time. The cathodic acupuncture electrodes working under an applied voltage of ∼3 V (with minimal damage to the living body) undergo effective electrochemical reactions in the acidic tumor area that produce sufficient H₂ locally to cause cancer cells to burst and die. Due to puncture positioning, the acidic tumor microenvironment and gas diffusion effect, the developed H₂ generation electrochemotherapy (H₂-ECT) strategy enables precise and large-scale tumor therapy, as demonstrated by in vivo treatment of diseased mice (glioma and breast cancers). Such green H₂-ECT is simple, highly efficient and minimally invasive, requiring no expensive medical equipment or nano materials and medication, and is therefore very promising for potential clinical applications.     

A new perspective on in vitro assessment method for evaluating quantum dot toxicity by using microfluidics technology

In this study, we demonstrate a new perspective on in vitro assessment method for evaluating quantum dot (QD) toxicity by using microfluidics technology. A new biomimetic approach, based on the flow exposure condition, was applied in order to characterize the cytotoxic potential of QD. In addition, the outcomes obtained from the flow exposure condition were compared to those of the static exposure condition. An in vitro cell array system was established that used an integrated multicompartmented microfluidic device to develop a sensitive flow exposure condition. QDs modified with cetyltrimethyl ammonium bromide∕trioctylphosphine oxide were used for the cytotoxicity assessment. The results suggested noticeable differences in the number of detached and deformed cells and the viability percentages between two different exposure conditions. The intracellular production of reactive oxygen species and release of cadmium were found to be the possible causes of QD-induced cytotoxicity, irrespective of the types of exposure condition. In contrast to the static exposure, the flow exposure apparently avoided the gravitational settling of particles and probably assisted in the homogeneous distribution of nanoparticles in the culture medium during exposure time. Moreover, the flow exposure condition resembled in vivo physiological conditions very closely, and thus, the flow exposure condition can offer potential advantages for nanotoxicity research.