Aldehyde N,N-dimethylhydrazone-based fluorescent substrate for peroxidase-mediated assays.

Numerous assays based on peroxidase activity have been developed for the detection of analytes due to the various optical peroxidase substrates. However, most substrates are sensitive to light and pH and are over-oxidized in the presence of excess H2O2. In this study, 2-((6-methoxynaphthalen-2-yl)methylene)-1,1-dimethylhydrazine (MNDH), a fluorescent peroxidase substrate prepared from naphthalene-based aldehyde N,N-dimethylhydrazone, was developed. MNDH showed quantitative fluorescence changes with respect to the H2O2 concentration in the presence of horseradish peroxidase (HRP), and the MNDH/HRP assay showed no changes in fluorescence caused by over-oxidation in the presence of excess H2O2. Further, MNDH was thermo- and photostable. Additionally, the assay could be operated over a considerably wide pH range, from acidic to neutral. Moreover, MNDH can be used to detect glucose quantitatively in human serum samples by using an enzyme cascade assay system.

Supra-threshold vibration applied to the foot soles enhances jump height under maximum effort.

Previous studies have shown that absence or reduction of cutaneous sensory feedback can diminish human motor performance under maximum effort. However, it has not been explored whether any appropriate intervention in the cutaneous sensory input can augment the output motor performance, particularly in motor tasks such as jumping that involve the kinematic chain of the entire body. Using shoes with active vibrating insoles, we applied mechanical vibration to the soles of 20 young and healthy adults and evaluated the change in the jump height and muscle activation using within-participants repeated measures. The noise-like vibration having an amplitude of 130% of the sensory threshold of each participant led to an average increase of 0.38 cm in the jump height (p = 0.008) and activation of the rectus femoris of the dominant leg (p = 0.011). These results indicate that application of a properly designed cutaneous stimulus to the soles, the distal end effectors of motor tasks, can augment the output performance by involving the prime movers distant from the end effector.

Gold nanoparticle-assisted delivery of small, highly structured RNA into the nuclei of human cells.

Previous studies have shown that functionalized gold nanoparticles (AuNPs) can be used as a general platform for loading and delivering DNA oligonucleotides and short hairpin RNA to living systems. Here, we report the ability of functionalized AuNP to deliver RNA aptamers into the nuclei of human cells. An in vitro-synthesized RNA aptamer specific to the ß-catenin protein was delivered into the HepG2 human cell line more efficiently via functionalized AuNP than liposome-based delivery, and resulted in nearly complete inhibition of ß-catenin binding to the p50 subunit of NF-κB in the nucleus. This inhibition led to repression of NF-κB p50-dependent transcription of CRP. Also, the ß-catenin aptamer in the nucleus led to down-regulation of ß-catenin-mediated transcriptional activity through the TCF complex and resulted in decrease in the levels of cyclin D, and c-myc mRNA by ~47% and ~57%, respectively. In addition, we used functionalized AuNP to deliver another RNA aptamer targeted to the p50 subunit of NF-κB into the A549 human cell line, and this was sufficient to induce apoptosis of the cells. Our findings demonstrate that AuNP GDS can be used to deliver small, highly structured RNA aptamers into the nucleus of human cells where they modulate the activity of transactivators by interacting with target proteins.

Delivery of shRNA using gold nanoparticle-DNA oligonucleotide conjugates as a universal carrier.

The efficient delivery of nucleic acids into mammalian cells is a central aspect of research involving cell biology and medical applications, including the clinical treatment of genetic disorders. We report an efficient small hairpin RNA (shRNA) delivery system that utilizes a single species of gold nanoparticle-DNA oligonucleotide conjugate (AuNP-DNA oligo) as a universal carrier. In vitro synthesized shRNA that is specific to the p53 gene was efficiently delivered into HEK293 and HeLa human cell lines using an AuNP-DNA oligo. The delivery resulted in an 80-90% knockdown of p53 expression. The same AuNP-DNA oligo was also efficient for the delivery of another shRNA, which is specific to the Mcl-1 gene, as well as the repression of MCL-1 expression. The knockdown efficiency of shRNA that was delivered using an AuNP-DNA oligo was comparable with that of a liposome-based shRNA delivery method. Our results offer an alternate delivery system for shRNA that can be used on any gene of interest.

Shoes with active insoles mitigate declines in balance after fatigue.

Fatigue can induce postural instability and even lead to falls. However, most current methods to delay or reduce fatigue require long preparatory time, or large and expensive equipment. We propose a convenient method to alleviate postural instability due to fatigue. We paid attention to that fatigue and aging share similar neurophysiological deterioration of sensory-motor function. Considering that stochastic resonance via sub-sensory mechanical vibration increases postural stability in the elderly, we propose that sub-sensory insole vibration reduces the negative effect of fatigue on postural control. We performed experiments with 21 young and healthy adult participants, and demonstrated that insole vibration compensates for the loss of balance ability due to fatigue. The sub-sensory insole vibration restored both the area of center of pressure and the complexity of the time series of the motor output after fatigue to the pre-fatigue levels. The insole units generating the vibration were completely concealed in shoes and controlled by a smart phone. This compact implementation contrasts with the cumbersome procedure of current solutions to fatigue-induced postural instability.

RGO-Coated TiO2 Microcones for High-Rate Lithium-Ion Batteries.

Reduced graphene oxide (RGO)-coated TiO2 microcones have been synthesized via simple anodization and cyclic voltammetry for use in lithium-ion batteries (LIBs). Microcones had a perpendicularly oriented hollow core, an anatase structure, and a high surface area, allowing higher capacity than other nanosized TiO2 structures. TiO2 has low electrical conductivity, leading to the limitation of fast charging and high capacity; however, this was improved by the application of an RGO coating in this work. As anode materials of LIB, the obtained RGO microcone showed a capacity of 157 mAh g-1 at 10C (fully charged within ∼360 s) and sustained 1000 cycles with only 0.02% capacity fading per cycle. The capacity was 1.5 times higher than that of conventional microcone. We speculated that the decrease in the charge-transfer resistance (R ct) played a crucial role in increasing the capacity with fast charging.

A highly sensitive gold nanoparticle-based colorimetric probe for pyrophosphate using a competition assay approach.

In this study, a mixture of [Zn(2)(1,3-bis[bis(2-pyridylmethyl)aminomethyl]benzene)](4+) ([Zn(2)(BBPAB)](4+)) and 11-mercaptoundecylphosphoric acid functionalized gold nanoparticles (Phos-AuNPs) is shown to be a highly sensitive colorimetric probe that can easily detect pyrophosphate (PPi) at less than 200 nM with the naked eye.

A simple, fast, and easy assay for transition metal-catalyzed coupling reactions using a paper-based colorimetric iodide sensor.

A paper-based colorimetric iodide sensor (PBCIS) that consists of filter paper treated with starch and an oxidant is developed. It has been employed as a protocol to obtain the extent of conversion of aryl iodides in C-C, C-N, C-O and C-S bond formations, including polymer-supported Heck reactions, by transition metal catalysts such as palladium, nickel and copper.

A colorimetric selective sensing probe for calcium ions with tunable dynamic ranges using cytidine triphosphate stabilized gold nanoparticles.

Cytidine triphosphate (CTP) stabilized-Au nanoparticles (CTP-AuNPs) allow a quantitative assay of Ca(2+) down to a concentration of 10(-4) M with high selectivity towards Ca(2+) ions over various metal ions including Mg(2+) and the detection range of this probe also can be easily tuned by changing the concentration of CTP.

Modulation of biological processes in the nucleus by delivery of DNA oligonucleotides conjugated with gold nanoparticles.

The development of a method that can efficiently deliver nucleic acids into the nucleus of living systems remains one of the key challenges for experimental and therapeutic use of nonbiological gene delivery agents. In the current study, we demonstrate a functionalized gold nanoparticle (AuNP) that can serve as a universal carrier for the delivery of DNA oligonucleotides (oligos) into the nucleus. We designed various types of DNA oligos to redirect alternative splicing of pre-mRNAs, such as MCL-1 and BCL-6, and to sequester transcriptional factors, including estrogen receptor α and p53. We successfully delivered the oligos into the nucleus, resulting in the targeted effects. In addition, injection of the antisense DNAs into a xenograft tumor in a mouse model system resulted in inhibited development of the tumor by redirecting the alternative splicing of the pre-mRNA. Our findings show that these nanoconjugates efficiently load and deliver antisense DNAs to redirect gene splicing or double-stranded DNAs to decoy gene transcription by transcriptional factors into mammalian cells and in vivo animals. Therefore, our lego-like AuNP gene delivery system can be used universally to control different biological processes by modulating nuclear gene expression events in living systems.

Inhibition of xenograft tumor growth in mice by gold nanoparticle-assisted delivery of short hairpin RNAs against Mcl-1L.

A prerequisite for the therapeutic use of small RNAs is the development of a method that can deliver them into animals. Previous studies have shown the capability of functionalized gold nanoparticles to serve as a general platform for loading and delivering DNA oligonucleotides and short hairpin RNAs (shRNAs) into cultured human cells. Here, we report the ability of the gold nanoparticle-assisted gene delivery system (AuNP-GDS) to deliver shRNA to a xenograft tumor in a mouse model. AuNP-GDS delivery of in vitro synthesized shRNA targeted to the Mcl-1L gene knocked down levels of Mcl-1L mRNA and protein by ~36% and ~26%, respectively, which were sufficient to induce apoptosis of the xenograft tumor cells and consequently inhibited the development of the tumor. We demonstrated that our lego-like AuNP-GDS, which can easily load and deliver shRNAs targeted to any gene of interest into living systems, can deliver shRNAs into xenograft tumors, leading to antitumor activity in an animal model.

A functionalized gold nanoparticles-assisted universal carrier for antisense DNA.

In this study, single-stranded DNA functionalized gold nanoparticles (AuNP GDS) were proved to be efficient gene delivery systems for oligo antisense DNAs specific to any gene of interest without affecting normal cell physiology.