Enhancing the optical path inside a capillary without sacrificing light intensity: application to a compact and highly sensitive photometer.

In order to increase the optical path and related sensitivity of photometers, multiple axial-reflection of parallel light-beam inside a capillary cavity is one of the most effective ways. However, there is a non-optimum trade-off between optical path and light intensity, e.g., smaller aperture on cavity mirror can increase multiple axial-reflection times (i.e., longer optical path) due to the lower cavity-loss, but it would also reduce coupling efficiency, light intensity, and related signal-to-noise ratio. Herein, an optical beam shaper, which is composed of two optical lenses with an apertured mirror, was proposed to focus the light beam (i.e., increasing coupling efficiency) without deteriorating beam parallelism and related multiple axial-reflection. Thus, by combining the optical beam shaper with a capillary cavity, large optical path enhancement (10-fold of capillary length) and high coupling efficiency (>65%) can be realized simultaneously, where the coupling efficiency was improved 50-fold. An optical beam shaper photometer (with a 7 cm long capillary) was fabricated and applied to detect water in ethanol with a detection limit of 12.5 ppm, which is 800-fold and 32∼80 fold lower than that of the commercial spectrometer (1 cm cuvette) and previous reports, respectively.

Carboxymethyl Chitosan and Gelatin Hydrogel Scaffolds Incorporated with Conductive PEDOT Nanoparticles for Improved Neural Stem Cell Proliferation and Neuronal Differentiation.

Tissue engineering scaffolds provide biological and physiochemical cures to guide tissue recovery, and electrical signals through the electroactive materials possess tremendous potential to modulate the cell fate. In this study, a novel electroactive hydrogel scaffold was fabricated by assembling poly(3,4-ethylenedioxythiophene) (PEDOT) nanoparticles on a carboxymethyl chitosan/gelatin (CMCS/Gel) composite hydrogel surface via in situ chemical polymerization. The chemical structure, morphology, conductivity, porosity, swelling rate, in vitro biodegradation, and mechanical properties of the prepared hydrogel samples were characterized. The adhesion, proliferation, and differentiation of neural stem cells (NSCs) on conductive hydrogels were investigated. The CMCS/Gel-PEDOT hydrogels exhibited high porosity, excellent water absorption, improved thermal stability, and adequate biodegradability. Importantly, the mechanical properties of the prepared hydrogels were similar to those of brain tissue, with electrical conductivity up to (1.52 ± 0.15) × 10-3 S/cm. Compared to the CMCS/Gel hydrogel, the incorporation of PEDOT nanoparticles significantly improved the adhesion of NSCs, and supported long-term cell growth and proliferation in a three-dimensional (3D) microenvironment. In addition, under the differentiation condition, the conductive hydrogel also significantly enhanced neuronal differentiation with the up-regulation of ß-tubulin III expression. These results suggest that CMCS/Gel-PEDOT hydrogels may be an attractive conductive substrate for further studies on neural tissue repair and regeneration.

Highly efficient genome editing via CRISPR-Cas9 in human pluripotent stem cells is achieved by transient BCL-XL overexpression.

Genome editing of human induced pluripotent stem cells (iPSCs) is instrumental for functional genomics, disease modeling, and regenerative medicine. However, low editing efficiency has hampered the applications of CRISPR-Cas9 technology in creating knockin (KI) or knockout (KO) iPSC lines, which is largely due to massive cell death after electroporation with editing plasmids. Here, we report that the transient delivery of BCL-XL increases iPSC survival by ∼10-fold after plasmid transfection, leading to a 20- to 100-fold increase in homology-directed repair (HDR) KI efficiency and a 5-fold increase in non-homologous end joining (NHEJ) KO efficiency. Treatment with a BCL inhibitor ABT-263 further improves HDR efficiency by 70% and KO efficiency by 40%. The increased genome editing efficiency is attributed to higher expressions of Cas9 and sgRNA in surviving cells after electroporation. HDR or NHEJ efficiency reaches 95% with dual editing followed by selection of cells with HDR insertion of a selective gene. Moreover, KO efficiency of 100% can be achieved in a bulk population of cells with biallelic HDR KO followed by double selection, abrogating the necessity for single cell cloning. Taken together, these simple yet highly efficient editing strategies provide useful tools for applications ranging from manipulating human iPSC genomes to creating gene-modified animal models.

The effect of glutamate-induced excitotoxicity on DNA methylation in astrocytes in a new in vitro neuron-astrocyte-endothelium co-culture system.

Glutamate-induced excitotoxicity is a contributer to many neurological diseases. Astrocytes may represent a new target for treating glutamate-induced excitotoxicity. However, the in vitro culture system that mimics the in vivo microenvironment is lacking. This study aimed to establish a new in vitro co-culture system including neurons, astrocytes, and endothelial cells (NAE), and to investigate the effect of glutamate-induced excitotoxicity on DNA methylation in astrocytes. A NAE co-culture method was created using a Transwell chamber, in which neurons were seeded on the bottom of the lower chamber, endothelial cells were plated on the top membrane, and astrocytes were plated on the bottom membrane of the insert. Glutamate-induced toxicity was induced using glutamate and glycine, and examined using immunofluorescence and lactate dehydrogenase release assay. Global methylation in astrocytes was analyzed, and the expression of DNMT1 and DNMT3a was examined using Western blot analysis. Glutamate treatment induced less neuronal damage in the NAE system compared with the control group in which neurons and astrocytes were cultured alone. Global DNA methylation was increased and the expression of DNMT1 and DNMT3a in astrocytes was increased after glutamate treatment, which was blocked by application of the NMDAR inhibitor MK-801 and the DNMT inhibitor 5-azaC from the endothelial cells. The in vitro ANE culture system is effective for studying glutamate-induced excitotoxicity, and may be used for testing the passage of drugs across the blood-brain barrier. Inhibition of DNA methylation in astrocytes may be a new therapeutic strategy for treating glutamate-induced excitotoxicity.

Correction: Liu, M., et al. Potent Effects of Flavonoid-Rich Extract from Rosa laevigata Michx Fruit against Hydrogen Peroxide-Induced Damage in PC12 Cells via Attenuation of Oxidative Stress, Inflammation and Apoptosis. Molecules 2014, 19, 11816-11832.

During the course of a review of our publication, an error in the title paper [...].

Genomic, expressional, protein-protein interactional analysis of Trihelix transcription factor genes in Setaria italia and inference of their evolutionary trajectory.

BACKGROUND: Trihelix transcription factors (TTF) play important roles in plant growth and response to adversity stress. Until now, genome-wide identification and analysis of this gene family in foxtail millet has not been available. Here, we identified TTF genes in the foxtail millet and its grass relatives, and characterized their functional domains. RESULTS: As to sequence divergence, TTF genes were previously divided into five subfamilies, I-V. We found that Trihelix family members in foxtail millet and other grasses mostly preserved their ancestral chromosomal locations during millions of years' evolution. Six amino acid sites of the SIP1 subfamily possibly were likely subjected to significant positive selection. Highest expression level was observed in the spica, with the SIP1 subfamily having highest expression level. As to the origination and expansion of the gene family, notably we showed that a subgroup of subfamily IV was the oldest, and therefore was separated to define a new subfamily O. Overtime, starting from the subfamily O, certain genes evolved to form subfamilies III and I, and later from subfamily I to develop subfamilies II and V. The oldest gene, Si1g016284, has the most structural changes, and a high expression in different tissues. What's more interesting is that it may have bridge the interaction with different proteins. CONCLUSIONS: By performing phylogenetic analysis using non-plant species, notably we showed that a subgroup of subfamily IV was the oldest, and therefore was separated to define a new subfamily O. Starting from the subfamily O, certain genes evolved to form other subfamilies. Our work will contribute to understanding the structural and functional innovation of Trihelix transcription factor, and the evolutionary trajectory.

Nε-(carboxymethyl)-lysine, White Matter, and Cognitive Function in Diabetes Patients.

OBJECTIVE: To study the relationship of Nε-(carboxymethyl)-lysine level (CML) with microstructure changes of white matter (WM), and cognitive impairment in patients with type 2 diabetes mellitus (T2DM) and to discuss the potential mechanism underlying T2DM-associated cognitive impairment. METHODS: The study was performed in T2DM patients (n=22) with disease course ≥5 years and age ranging from 65 to 75 years old. A control group consisted of 25 sex- and age-matched healthy volunteers. Fractional anisotropy (FA) of several WM regions was analyzed by diffusion tensor imaging scan. Plasma CML levels were measured by enzyme-linked immunosorbent assay, and cognitive function was assessed by Mini-Mental State Examination and Montreal cognitive assessment (MoCA). RESULTS: The total Mini-Mental State Examination score in the patient group (25.72±3.13) was significantly lower than the control group (28.16±2.45) (p<0.05). In addition, the total MoCA score in the patient group (22.15±3.56) was significantly lower than the control group 25.63±4.12) (p<0.01). In the patient group, FA values were significantly decreased in the corpus callosum, cingulate fasciculus, inferior fronto-occipital fasciculus, parietal WM, hippocampus, and temporal lobes relative to corresponding regions of healthy controls (p<0.05). Plasma CML level was negatively correlated with average FA values in the global brain (r=-0.58, p<0.01) and MoCA scores (r=-0.47, p<0.05). CONCLUSIONS: In T2DM, WM microstructure changes occur in older patients, and elevations in CML may play a role in the development of cognitive impairment.

A new method for evaluation of motor injury after acute brain ischemic damage.

PURPOSE: Motor impairment is an important index for assessing the extent of brain injury. The present study uses a new method, the movement capture analysis (MOCA) system, for assessing motor damage after acute ischemia. MATERIALS AND METHODS: Forty rats were divided into four groups: standard ischemia, sham-operated, Dizocilpine (MK-801), and Ginkgo biloba extract (GBE) groups. Brain ischemia was induced using the temporary right middle cerebral artery occlusion model. Longa score and MOCA were used to assess motor injury one day after ischemia. Infarct volume was delineated with 2% 2,3,5-triphenyltetrazolium chloride (TTC) staining. The correlation of infarct volume with Longa score and MOCA data was calculated. RESULTS: Compared with the sham-operated group (0.10 ± 0.31), Longa scores of MK-801 (2.33 ± 0.73), GBE (1.80 ± 0.58), and standard (2.88 ± 0.83) groups showed a statistical difference (p < 0.05); however, it was unable to discern the difference between MK-801 and standard groups. MOCA was able to clearly discern the differences in motor disparity among the four groups, standard (1.00 ± 0.19), sham-operated group (0.17 ± 0.02), MK-801 (0.79 ± 0.08), GBE (0.38 ± 0.05) (p < 0.05). Both MK-801 (18.03 ± 0.96%) and GBE (10.82 ± 1.93%) treatment reduced infarct size compared with the standard ischemia group (25.88 ± 1.16%) (p < 0.05). The MOCA data showed a more significant correlation with infarct size than Longa score (r = 0.85:0.53). CONCLUSIONS: MOCA system proved to be more sensitive than the Longa score. It may potentially be more accurate method for behavioral evaluation in clinical trials.

[Research on the dynamics of ion debris from Sn plasma by use of dual laser pulses].

Extreme ultraviolet lithography is one of the most promising technologies on the next generation of high-capacity integrated circuit manufacturing. However, techniques for ion debris mitigation have to be considered in the application of extreme ultraviolet source for lithography. In our paper the dynamics of ion debris from Sn plasma by using dual ns laser pulses were investigated. The results show that debris from plasma greatly depends on the energy of pre-pulse and the delay time between the two laser pulses. The energy of Sn ions debris was efficiently mitigated from 2. 47 to 0. 40 keV in the case of dual laser pulses, up to 6. 1 times lower than that by using single laser pulse. We also found that Sn ions debris can be mitigated at all angles by using the dual laser pulses method.

Roseomonas sediminicola sp. nov., isolated from fresh water.

A Gram-stain negative, strictly aerobic, non-motile, non-spore-forming, and rod-shaped bacterial strain designated FW-3(T) was isolated from fresh water and its taxonomic position was investigated by using a polyphasic approach. Strain FW-3(T) was found to grow at 10-37 °C and at pH 7.0 in the absence of NaCl on nutrient agar. On the basis of 16S rRNA gene sequence similarity, strain FW-3(T) was shown to belong to the family Acetobacteraceae and to be related to Roseomonas lacus TH-G33(T) (97.2 % sequence similarity) and Roseomonas terrae DS-48(T) (96.4 %). The G+C content of the genomic DNA was determined to be 68.0 %. The major menaquinone was determined to be Q-10 and the major fatty acids were identified as summed feature 7 (comprising C18:1 ω9c/ω12t/ω7c as defined by the MIDI system; 55.4 %), and C18:1 2OH (29.8 %). DNA and chemotaxonomic data supported the affiliation of strain FW-3(T) to the genus Roseomonas. Strain FW-3(T) could be differentiated genotypically and phenotypically from the recognized species of the genus Roseomonas. The novel isolate therefore represents a novel species, for which the name Roseomonas sediminicola sp. nov. is proposed, with the type strain FW-3(T) (=KACC 16616(T) = JCM 18210(T)).

Preparation of przewalskinic acid A from salvianolic acid B using a crude enzyme from an Aspergillus oryzae strain.

Przewalskinic acid A is a rare, water-soluble, and highly biologically active ingredient found, thus far, only in the Salvia przewalskii Maxim herb; however, the content in S. przewalskii herb is very low. In order to obtain useful quantities of przewalskinic acid A, the biotransformatin of salvianolic acid B from Salvia miltiorrhiza root (danshen in Chinese) into przewalskinic acid A was studied using a crude enzyme produced from Aspergillus oryzae D30s strain. The crude enzyme from the A. oryzae strain hydrolyzed salvianolic acid B into przewalskinic acid A and danshensu. The preparation afforded 31.3 g przewalskinic acid A (91.0 % purity) and 13.1 g danshensu (95 % purity) from 75 g salvianolic acid B. The preparation of przewalskinic acid A was therefore very successful with a yield of over 86 %, but the yield of danshensu was only 33 %. The product przewalskinic acid A was identified using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) and NMR.

Potent effects of flavonoid-rich extract from Rosa laevigata Michx fruit against hydrogen peroxide-induced damage in PC12 cells via attenuation of oxidative stress, inflammation and apoptosis.

Oxidative stress-induced neuronal death has an important role in the pathogenesis of neurodegenerative disorders. The effects and mechanisms of action of the total flavonoids (TFs) from Rosa laevigata Michx fruit against hydrogen peroxide (H2O2)-induced oxidative injury in PC12 cells were investigated in this study. The results demonstrated that the TFs protected against cell apoptosis, DNA and mitochondrial damage caused by H2O2 based on single cell gel electrophoresis, in situ terminal deoxynucleotidyltransferase dUTP nick end labeling (TUNEL), flow cytometry and transmission electron microscope (TEM) assays. In addition, the TFs notably decreased cytochrome C release from mitochondria into the cytosol and intracellular Ca2+ levels, and diminished intracellular generation of reactive oxygen species (ROS). Furthermore, the TFs inhibited the phosphorylation levels of JNK, ERK and p38 MAPK as well as down-regulated the expressions of IL-1, IL-6, TNF-α, Fas, FasL, CYP2E1, Bak, caspase-3, caspase-9, p53, COX-2, NF-κB, AP-1, and up-regulated the expressions of Bcl-2 and Bcl-xl. In conclusion, these results suggest that the TFs from R. laevigata Michx fruit show good effects against H2O2-induced oxidative injury in PC12 cells by adjusting oxidative stress, and suppression of apoptosis and inflammation, and could be developed as a potential candidate to prevent oxidative stress in the future.

[The LIBS experiment condition optimization of alloy steel].

Laser induced plasma spectroscopy of alloy steel was produced by Nd : YAG pulsed laser at 1 064 nm, and the spectral signal was detected by high resolution and width controlled ICCD. Several Fe atomic spectral lines such as 404.581, 414.387, 427.176 and 438.355 nm were chosen for analysis, and the effects of different experimental parameters on LIBS spectral signal intensity were investigated. It is shown that the experimental parameters such as pulse energy, laser focus location and laser delay time have great influence on the LIBS signal. LIBS signals with high spectral intensity and signal-background ratio (SBR) as well as the optimum experiment conditions were obtained by optimizing these experiment parameters so as to make composition analysis of the alloy steel.

Bioelectronic medicine potentiates endogenous NSCs for neurodegenerative diseases.

Neurodegenerative diseases (NDs) are commonly observed and while no therapy is universally applicable, cell-based therapies are promising. Stem cell transplantation has been investigated, but endogenous neural stem cells (eNSCs), despite their potential, especially with the development of bioelectronic medicine and biomaterials, remain understudied. Here, we compare stem cell transplantation therapy with eNSC-based therapy and summarize the combined use of eNSCs and developing technologies. The rapid development of implantable biomaterials has resulted in electronic stimulation becoming increasingly effective and decreasingly invasive. Thus, the combination of bioelectronic medicine and eNSCs has substantial potential for the treatment of NDs.

Electrical stimulation enhances the neuronal differentiation of neural stem cells in three-dimensional conductive scaffolds through the voltage-gated calcium ion channel.

Electrical stimulation (ES) or electroconductive scaffold has been proved to have the positive effects on the behavior of neural stem cells (NSCs). We previously developed a novel three-dimensional conductive composite scaffold of poly (3, 4-ethylenedioxythiophen)/chitosan/gelatin (PEDOT/Cs/Gel) for neural tissue engineering. In the present study, we further studied the effect of three-dimensional conductive scaffolds combined with ES on the neuronal differentiation of NSCs. The sandwiched ES device was designed to apply single-phase pulse voltage on NSCs cultured in conductive scaffold for 7 days (4 h/day). Proliferation and differentiation related proteins and genes were analyzed by immunofluorescence staining and RT-qPCR. The role of voltage-gated ion channels (VGICs) in regulating NSCs' neuronal differentiation by ES was investigated in presence of ion channels blockers. The results of protein and gene expression indicated that ES not only promoted the proliferation of NSCs cultured in the conductive scaffold, but also enhanced the differentiation of NSCs into neurons. Especially, the voltage-gated calcium channel (Cav2+) played an important role in the neuronal differentiation of NSCs under ES. Our findings demonstrated that ES combined with three-dimensional conductive scaffolds would be a promising strategy to regulate the neuronal differentiation of NSCs for neural regeneration.

Triggering and guiding high-voltage discharge in air by single and multiple femtosecond filaments.

The abilities to trigger and guide high-voltage discharge by using single and multiple filaments (MFs) are experimentally studied. It is shown that the discharge voltage threshold can be reduced significantly in both regimes of single and MF; however, the MF does not gain a larger reduction than a single filament. This behavior of the MF is attributed to the single discharge path rather than simultaneous multiple ones as one might expect during the discharge process.

Blocking glutamate-mediated signalling inhibits human melanoma growth and migration.

Glutamate is an excitatory neurotransmitter that has been shown to regulate the proliferation, migration and survival of neuronal progenitors in the central nervous system through its action on metabotropic and ionotropic glutamate receptors (GluRs). Antagonists of ionotropic GluRs have been shown to cause a rapid and reversible change in melanocyte dendritic morphology, which is associated with the disorganization of actin and tubulin microfilaments in the cytoskeleton. Intracellular expression of microtubule-associated protein (MAP) 2a affects the assembly, stabilization and bundling of microtubules in melanoma cells; stimulates the development of dendrites; and suppresses melanoma cell migration and invasion. In this study, we investigated the relationship between glutamate-mediated signalling and microtubules, cell dendritic morphology and melanoma cell motility. We found that metabotropic GluR1 and N-methyl-d-aspartate receptor antagonists increased dendritic branching and inhibited the motility, migration and proliferation of melanoma cells. We also demonstrated that the invasion and motility of melanoma cells are significantly inhibited by the combination of increased expression of MAP2a and either metabotropic GluR1 or N-methyl-d-aspartate receptor antagonists. Moreover, the blockade of glutamate receptors inhibited melanoma growth in vivo. Collectively, these results demonstrate the importance of glutamate signalling in human melanoma and suggest that the blockade of glutamate receptors is a promising novel therapy for treating melanoma.

Enhanced in vitro antitumor efficacy and strong anti-cell-migration activity of a hydroxycamptothecin-encapsulated magnetic nanovehicle.

A 10-hydroxycamptothecin-encapsulated magnetic nanovehicle (HEMN) was fabricated by coencapsulating Fe(3)O(4) nanoparticles and 10-hydroxycamptothecin (HCPT) into a micelle core self-assembled from the amphiphilic copolymer methoxy-poly(ethylene glycol)-poly(d,l-lactide-co-glycolide) through a facile dialysis method. A satisfactory drug-loading content of (9.03 ± 0.67) % and a relatively high encapsulation efficiency of (53.52 ± 6.46) % were achieved. In vitro drug release was performed by membrane dialysis and a pH-dependent release behavior was observed. In comparison with free HCPT dissolved in dimethylsulfoxide, HEMNs showed a greatly improved in vitro antitumor efficacy against three different human cancer cell lines-HeLa, A549, and HepG2-and lower IC(50) values were measured. The mechanism of cell death was investigated, and it was clearly demonstrated that the apoptosis process was triggered. An in vitro wound-healing assay and a transwell assay indicated that HEMNs exerted much stronger activity in inhibiting HeLa cell migration. The cellular uptake of HEMNs in a desired area can be significantly enhanced by an external magnetic field. These results demonstrate HCPT-encapsulated magnetic nanovehicles might have important potential in clinical applications for inhibiting tumor metastasis and for targeted drug delivery.

Synthesis and characterization of protocatechuic acid grafted carboxymethyl chitosan with oxidized sodium alginate hydrogel through the Schiff's base reaction.

Excessive accumulation of free radicals is closely related to the occurrence and development of various neurodegenerative diseases. In this study, a novel protocatechuic acid grafted carboxymethyl chitosan with oxidized sodium alginate (PCA-g-CMCS/OSA) hydrogel was developed to maintain the oxidation-antioxidation balance activities. By optimizing the pH-soluble range (pH > 6.4) of CMCS, PCA was grafted onto CMCS skeleton via EDC/NHS, and then conjugated with aldehyde group of OSA to form Schiff's base hydrogel at physiological temperature. The gelation time can be adjusted rapidly within 1-3 min by controlling the content of OSA. The shaped hydrogel exhibited porous network structure with high porosity (>90 %), swelling ratio (2000-3000 %) and rheological property, which is beneficial to cell growth and proliferation. The conjugates preserved excellent DPPH and ABTS radicals scavenging abilities and adequate biodegradability within 5 weeks. Moreover, with the release of PCA monomer due to degradation of the PCA-g-CMCS/OSA, the hydrogel also exhibited excellent biocompatibility and protective effect on H2O2-induced oxidative damage in PC12 cells. These results suggested that the PCA-g-CMCS/OSA hydrogel would appear to be a more attractive candidate for potential biomedical applications such as antioxidant drug release and tissue engineering implant material.

Learning Asynchronous Boolean Networks From Single-Cell Data Using Multiobjective Cooperative Genetic Programming.

Recent advances in high-throughput single-cell technologies provide new opportunities for computational modeling of gene regulatory networks (GRNs) with an unprecedented amount of gene expression data. Current studies on the Boolean network (BN) modeling of GRNs mostly depend on bulk time-series data and focus on the synchronous update scheme due to its computational simplicity and tractability. However, such synchrony is a strong and rarely biologically realistic assumption. In this study, we adopt the asynchronous update scheme instead and propose a novel framework called SgpNet to infer asynchronous BNs from single-cell data by formulating it into a multiobjective optimization problem. SgpNet aims to find BNs that can match the asynchronous state transition graph (STG) extracted from single-cell data and retain the sparsity of GRNs. To search the huge solution space efficiently, we encode each Boolean function as a tree in genetic programming and evolve all functions of a network simultaneously via cooperative coevolution. Besides, we develop a regulator preselection strategy in view of GRN sparsity to further enhance learning efficiency. An error threshold estimation heuristic is also proposed to ease tedious parameter tuning. SgpNet is compared with the state-of-the-art method on both synthetic data and experimental single-cell data. Results show that SgpNet achieves comparable inference accuracy, while it has far fewer parameters and eliminates artificial restrictions on the Boolean function structures. Furthermore, SgpNet can potentially scale to large networks via straightforward parallelization on multiple cores.