Control of Neuronal Survival and Development Using Conductive Diamond.

This study demonstrates the control of neuronal survival and development using nitrogen-doped ultrananocrystalline diamond (N-UNCD). We highlight the role of N-UNCD in regulating neuronal activity via near-infrared illumination, demonstrating the generation of stable photocurrents that enhance neuronal survival and neurite outgrowth and foster a more active, synchronized neuronal network. Whole transcriptome RNA sequencing reveals that diamond substrates improve cellular-substrate interaction by upregulating extracellular matrix and gap junction-related genes. Our findings underscore the potential of conductive diamond as a robust and biocompatible platform for noninvasive and effective neural tissue engineering.

Periodic Rayleigh streaming vortices and Eckart flow arising from traveling-wave-based diffractive acoustic fields.

Recent studies have demonstrated that periodic time-averaged acoustic fields can be produced from traveling surface acoustic waves (SAWs) in microfluidic devices. This is caused by diffractive effects arising from a spatially limited transducer. This permits the generation of acoustic patterns evocative of those produced from standing waves, but instead with the application of a traveling wave. While acoustic pressure fields in such systems have been investigated, acoustic streaming from diffractive fields has not. In this work we examine this phenomenon and demonstrate the appearance of geometry-dependent acoustic vortices, and demonstrate that periodic, identically rotating Rayleigh streaming vortices result from the imposition of a traveling SAW. This is also characterized by a channel-spanning flow that bridges between adjacent vortices along the channel top and bottom. We find that the channel dimensions determine the types of streaming that develops; while Eckart streaming has been previously presumed to be a distinguishing feature of traveling-wave actuation, we show that Rayleigh streaming vortices also results. This has implications for microfluidic actuation, where traveling acoustic waves have applications in microscale mixing, separation, and patterning.

Single-Step Fabrication Method toward 3D Printing Composite Diamond-Titanium Interfaces for Neural Applications.

Owing to several key attributes, diamond is an attractive candidate material for neural interfacing electrodes. The emergence of additive-manufacturing (AM) of diamond-based materials has addressed multiple challenges associated with the fabrication of diamond electrodes using the conventional chemical vapor deposition (CVD) approach. Unlike the CVD approach, AM methods have enabled the deposition of three-dimensional diamond-based material at room temperature. This work demonstrates the feasibility of using laser metal deposition to fabricate diamond-titanium hybrid electrodes for neuronal interfacing. In addition to exhibiting a high electrochemical capacitance of 1.1 mF cm-2 and a low electrochemical impedance of 1 kΩ cm2 at 1 kHz in physiological saline, these electrodes exhibit a high degree of biocompatibility assessed in vitro using cortical neurons. Furthermore, surface characterization methods show the presence of an oxygen-rich mixed-phase diamond-titanium surface along the grain boundaries. Overall, we demonstrated that our unique approach facilitates printing biocompatible titanium-diamond site-specific coating-free conductive hybrid surfaces using AM, which paves the way to printing customized electrodes and interfacing implantable medical devices.

Mechanisms and Applications of Neuromodulation Using Surface Acoustic Waves-A Mini-Review.

The study of neurons is fundamental for basic neuroscience research and treatment of neurological disorders. In recent years ultrasound has been increasingly recognized as a viable method to stimulate neurons. However, traditional ultrasound transducers are limited in the scope of their application by self-heating effects, limited frequency range and cavitation effects during neuromodulation. In contrast, surface acoustic wave (SAW) devices, which are producing wavemodes with increasing application in biomedical devices, generate less self-heating, are smaller and create less cavitation. SAW devices thus have the potential to address some of the drawbacks of traditional ultrasound transducers and could be implemented as miniaturized wearable or implantable devices. In this mini review, we discuss the potential mechanisms of SAW-based neuromodulation, including mechanical displacement, electromagnetic fields, thermal effects, and acoustic streaming. We also review the application of SAW actuation for neuronal stimulation, including growth and neuromodulation. Finally, we propose future directions for SAW-based neuromodulation.

Highly multiplexed quantifications of 299 somatic mutations in colorectal cancer patients by automated MALDI-TOF mass spectrometry.

BACKGROUND: Detection of somatic mutations in tumor tissues helps to understand tumor biology and guide treatment selection. Methods such as quantitative PCR can analyze a few mutations with high efficiency, while next generation sequencing (NGS) based methods can analyze hundreds to thousands of mutations. However, there is a lack of cost-effective method for quantitatively analyzing tens to a few hundred mutations of potential biological and clinical significance. METHODS: Through a comprehensive database and literature review we selected 299 mutations associated with colorectal cancer. We then designed a highly multiplexed assay panel (8-wells covering 299 mutations in 109 genes) based on an automated MADLI-TOF mass spectrometry (MS) platform. The multiplex panel was tested with a total of 319 freshly frozen tissues and 92 FFPE samples from 229 colorectal cancer patients, with 13 samples also analyzed by a targeted NGS method covering 532 genes. RESULTS: Multiplex somatic mutation panel based on MALDI-TOF MS detected and quantified at least one somatic mutation in 142 patients, with KRAS, TP53 and APC being the most frequently mutated genes. Extensive validation by both capillary sequencing and targeted NGS demonstrated high accuracy of the multiplex MS assay. Out of 35 mutations tested with plasmid constructs, sensitivities of 5 and 10% mutant allele frequency were achieved for 19 and 16 mutations, respectively. CONCLUSIONS: Automated MALDI-TOF MS offers an efficient and cost-effective platform for highly multiplexed quantitation of 299 somatic mutations, which may be useful in studying the biological and clinical significance of somatic mutations with large numbers of cancer tissues.

Dynamics of Plasma EGFR T790M Mutation in Advanced NSCLC: A Multicenter Study.

BACKGROUND: Droplet digital polymerase chain reaction (ddPCR) is an emerging technology for quantitative cell-free DNA oncology applications. However, a ddPCR assay for the epidermal growth factor receptor (EGFR) p.Thr790Met (T790M) mutation suitable for clinical use remains to be established with analytical and clinical validations. OBJECTIVE: We aimed to develop and validate a new ddPCR assay to quantify the T790M mutation in plasma for monitoring and predicting the progression of advanced non-small-cell lung cancer (NSCLC). METHODS: Specificity of the ddPCR assay was evaluated with genomic DNA samples from healthy individuals. The inter- and intraday variations of the assay were evaluated using mixtures of plasmid DNA containing wild-type EGFR and T790M mutation sequences. We assessed the clinical utility of the T790M assay in a multicenter prospective study in patients with advanced NSCLC receiving tyrosine kinase inhibitor (TKI) treatment by analyzing longitudinal plasma DNA samples. RESULTS: We set the criteria for a positive call when the following conditions were satisfied: (1) T790M mutation frequency > 0.098% (3 standard deviations above the background signal); (2) at least two positive droplets in duplicate ddPCR reactions. Among the 62 patients with advanced NSCLC exhibiting resistance to TKI treatment, 15 had one or more serial plasma samples that tested positive for T790M. T790M mutation was detected in the plasma as early as 205 days (median 95 days) before disease progression, determined by imaging analysis. Plasma T790M concentrations also correlated with intervention after disease progression. CONCLUSIONS: We developed a ddPCR assay to quantify the T790M mutation in plasma. Quantification of longitudinal plasma T790M mutation may allow noninvasive assessment of drug resistance and guide follow-up treatment in TKI-treated patients with NSCLC. TRIAL REGISTRATION: Clinical Trials.gov identifier: NCT02804100.

Effect of EDTA and citric acid on absorption of heavy metals and growth of Moso bamboo.

The effect of EDTA and citric acid on accumulation, toxicity of heavy metals (Cu, Zn, Cd, and Pb), and growth of Moso bamboo was investigated in current experiment. The availability of heavy metals in soil and its uptake by plants has indicated toxicity. The results revealed that EDTA and citric acid has reduced biomass of Moso bamboo but non-significant difference in biomass was observed compared with control. Application of EDTA (10 mmol kg-1) has significantly improved copper (Cu) by 56.5 and 84.9% in roots and above ground parts of plants. Application of EDTA (10 mmol kg-1) has significantly enhanced lead (Pb) by 51.8 and 210.8% in roots and above ground parts of Moso bamboo. Furthermore, treatment of EDTA has significantly improved activities of water-soluble Cd, Cu, and Pb in soil by 98.9, 70.1, and 73.1 times compared with control. In case of contents of diethylenetriaminepentaacetic acid (DTPA)-extractable metals, the treatment of EDTA (10 mmol kg-1) has produced maximum increase of 244.5 mg kg-1 Zn and 157.9 mg kg-1 Pb, respectively. It is concluded that effect of EDTA was superior compared with citric acid for improvement of phytoremediation potential of Moso bamboo.

Regulatory effects of saponins from Panax japonicus on colonic epithelial tight junctions in aging rats.

BACKGROUND: Saponins from Panax japonicus (SPJ) are the most abundant and main active components of P. japonicus, which replaces ginseng roots in treatment for many kinds of diseases in the minority ethnic group in China. Our previous studies have demonstrated that SPJ has the effects of anti-inflammation through the mitogen-activated protein kinase (MAPK) and nuclear factor kappa B (NF-κB) signaling pathways. The present study was designed to investigate whether SPJ can modulate intestinal tight junction barrier in aging rats and further to explore the potential mechanism. METHODS: Aging rats had been treated with different doses (10 mg/kg, 30 mg/kg, and 60 mg/kg) of SPJ for 6 mo since they were 18 mo old. After the rats were euthanized, the colonic samples were harvested. Levels of tight junctions (claudin-1 and occludin) were determined by immunohistochemical staining. Levels of proinflammatory cytokines (interleukin-1ß and tumor necrosis factor-α) were examined by Western blot. NF-κB and phosphorylation of MAPK signaling pathways were also determined by Western blot. RESULTS: We found that SPJ increased the expression of the tight junction proteins claudin-1 and occludin in the colon of aging rats. Treatment with SPJ decreased the levels of interleukin-1ß and tumor necrosis factor-α, reduced the phosphorylation of three MAPK isoforms, and inhibited the expression of NF-κB in the colon of aging rats. CONCLUSION: The studies demonstrated that SPJ modulates the damage of intestinal epithelial tight junction in aging rats, inhibits inflammation, and downregulates the phosphorylation of the MAPK and NF-κB signaling pathways.

Cadmium-induced oxidative stress, response of antioxidants and detection of intracellular cadmium in organs of moso bamboo (Phyllostachys pubescens) seedlings.

Moso bamboo (Phyllostachys pubescens (Pradelle) Mazel ex J.Houz.) is recognized as a potential phytoremediation plant due to its huge biomass and high tolerance to environmental stresses. The objectives of this study were to investigate mechanism related to cadmium (Cd) tolerance and to evaluate Cd accumulation capacity of moso bamboo. The results of the pot experiment showed that Cd accumulation by bamboo increased with increasing the Cd levels in soil and the values in stem ranged from 28.51 to 132.13 mg kg(-1). Meanwhile chlorophyll in leaves and total biomass showed a decreasing trend. The bioaccumulation factors (BAF) for roots and stem in all the treatments were more than 1.0 and the translocation factor (TF) ranged from 0.70 to 1.06. In hydroponics experiment, the concentrations of malondialdehyde (MDA) in the leaves were significantly increased in Cd treated plants as compared with control. The activities of superoxide dismutase (SOD) and peroxidase (POD) were enhanced at initial stage and then decreased consistently with the increase of Cd addition. The proline concentrations were also increased due to the presence of Cd, particularly at 25 µM Cd treatment. According to TEM-EDX analysis, the cytoplasm was the main site for accumulation of Cd in moso bamboo. On the basis of overall results, it is suggested that moso bamboo could be successfully used for the remediation of low Cd (no more than 5 mg kg(-1)) contaminated soils.

Copper induced oxidative stresses, antioxidant responses and phytoremediation potential of Moso bamboo (Phyllostachys pubescens).

Moso bamboo is recognized as phytoremediation plant due to production of huge biomass and high tolerance in stressed environment. Hydroponics and pot experiments were conducted to investigate mechanism of copper tolerance and to evaluate copper accumulation capacity of Moso bamboo. In hydroponics experiment there was non significant variation in MDA contents of leaves compared with control. SOD and POD initially indicated enhancing trend with application of 5 µM Cu and then decreased consistently with application of 25 and 100 µM Cu. Application of each additional increment of copper have constantly enhanced proline contents while maximum increase of proline was observed with application of 100 µM copper. In pot experiment chlorophyll and biomass initially showed increasing tendency and decreased gradually with application of each additional increment of Cu. Normal growth of Moso bamboo was observed with application of 100 mg kg(-1) copper. However, additional application of 300 or 600 mg kg(-1) copper had significantly inhibited growth of Moso bamboo. The concentration of Cu in Moso bamboo has attained the levels of 340, 60, 23 mg kg(-1) in roots, stems and leaves respectively. The vacuoles were the main organs which accumulated copper and reduced toxicity of copper as studied by TEM-DEX technology.

Effect of Zn stresses on physiology, growth, Zn accumulation, and chlorophyll of Phyllostachys pubescen s.

The effects of Zinc (Zn) on lipid peroxidation, antioxidative enzymes, growth, Zn accumulation, and leaf chlorophyll of Phyllostachys pubescens (Pradelle) Mazel ex J.Houz. were investigated in two greenhouse experiments. Hydroponics experiment with Zn application of 0, 20, 100, and 400 µM revealed that lower concentration of Zn in solution led to increased malondialdehyde (MDA) and proline contents but inhibited SOD activity in all treatments. P. pubescens had showed strong ability to accumulate Zn in stems and reached maximum level at 100 µM with 7.91-fold increase compared with control. In pot experiment, treatment with Zn ranged from 0, 200, 400, 800, 1,600, to 3,200 mg kg(-1). Application of 800 mg kg(-1) revealed 116, 24.6, and 28.3 times increase in Zn concentration of roots, stems, and leaves, respectively. Growth and chlorophyll contents of plants in pots were better promoted at 400 mg kg(-1) Zn, with 60.5 and 30.9 % enhanced roots and shoot compared with control. The bioaccumulation factor (BAF) was in the sequence of stem > roots > leaves. The translocation factor (TF) of stem was higher than leaves.

Lead accumulation and tolerance of Moso bamboo (Phyllostachys pubescens) seedlings: applications of phytoremediation.

A hydroponics experiment was aimed at identifying the lead (Pb) tolerance and phytoremediation potential of Moso bamboo (Phyllostachys pubescens) seedlings grown under different Pb treatments. Experimental results indicated that at the highest Pb concentration (400 µmol/L), the growth of bamboo seedlings was inhibited and Pb concentrations in leaves, stems, and roots reached the maximum of 148.8, 482.2, and 4282.8 mg/kg, respectively. Scanning electron microscopy revealed that the excessive Pb caused decreased stomatal opening, formation of abundant inclusions in roots, and just a few inclusions in stems. The ultrastructural analysis using transmission electron microscopy revealed that the addition of excessive Pb caused abnormally shaped chloroplasts, disappearance of endoplasmic reticulum, shrinkage of nucleus and nucleolus, and loss of thylakoid membranes. Although ultrastructural analysis revealed some internal damage, even the plants exposed to 400 µmol/L Pb survived and no visual Pb toxicity symptoms such as necrosis and chlorosis were observed in these plants. Even at the highest Pb treatment, no significant difference was observed for the dry weight of stem compared with controls. It is suggested that use of Moso bamboo as an experimental material provides a new perspective for remediation of heavy metal contaminated soil owing to its high metal tolerance and greater biomass.

Effect of Zn toxicity on root morphology, ultrastructure, and the ability to accumulate Zn in Moso bamboo (Phyllostachys pubescens).

The effects of zinc (Zn) on seed germination and growth of Moso bamboo (Phyllostachys pubescens) were investigated. Under zinc stress, the seed germination rate did not show significant difference from that of the control. Hydroponics experiments indicated that Moso bamboo had a strong ability to accumulate Zn in the shoot and it reached its maximum value in the shoot at 100 µM Zn. The root Zn concentration ranged from 2,329.29 to 8,642.51 mg kg(-1), with the root Zn concentration at 10 µM Zn being 58.23 times that of the control. The root morphology parameters slightly increased at the lower Zn treatments, while growth restriction was evident at higher Zn treatments. Root ultrastructural studies revealed that the cell structure, root tips, and organelles were significantly changed under Zn stress as compared to those of the control. Some abnormalities were evident in the cell walls, vacuoles, mitochondria, plasmalemma, tonoplast, and xylem parenchyma of root cells. While Moso bamboo seems a suitable candidate for phytoremediation, its metal remediation ability should be further explored in future investigations.

Effect of copper toxicity on root morphology, ultrastructure, and copper accumulation in Moso bamboo (Phyllostachys pubescens).

A hydroponic culture experiment was conducted to study the effect of copper toxicity on root morphology, ultrastructure, and copper accumulation in Moso bamboo (Phyllostachys pubescens). Root ultrastructure of Moso bamboo was studied by transmission electron microscopy and scanning electron microscopy. Application of 200 µM Cu resulted in an accumulation of 810 mg kg(-1) dry weight and 91 mg kg(-1) dry weight Cu in roots and shoots, respectively. The majority of the plants did not survive the application of 400 µM Cu. Biomass production declined consistently with application of each additional increment of Cu. Root growth was more severely inhibited than shoot growth. Cu adversely affected the root morphology of the plants, however, root surface area and number of root tips increased slightly at low levels of Cu. Root cell ultrastructure and organelles changed significantly under Cu stress, in particular, cell walls, mitochondria, and xylem parenchyma were affected.