Project description:Micro RNA (miR) are regulatory non-coding RNA molecules, which contain a small number of nucleotides ~18-28 nt. There are many various miR sequences found in plants and animals that perform important functions in developmental, metabolic, and disease processes. miRs can bind to complementary sequences within mRNA molecules thus silencing mRNA. Other functions include cardiovascular and neural development, stem cell differentiation, apoptosis, and tumors. In tumors, some miRs can function as oncogenes, others as tumor suppressors. Levels of certain miR molecules reflect cellular events, both normal and pathological. Therefore, miR molecules can be used as biomarkers for disease diagnosis and prognosis. One of these promising molecules is miR-21, which can serve as a biomarker with high potential for early diagnosis of various types of cancer. Here, we present a novel design of miR detection and demonstrate its efficacy on miR-21. The design employs emissive properties of DNA-silver nanoclusters (DNA/AgNC). The detection probe is designed as a hairpin DNA structure with one side of the stem complimentary to miR molecule. The binding of target miR-21 opens the hairpin structure, dramatically modulating emissive properties of AgNC hosted by the C12 loop of the hairpin. "Red" fluorescence of the DNA/AgNC probe is diminished in the presence of the target miR. At the same time, "green" fluorescence is activated and its intensity increases several-fold. The increase in intensity of "green" fluorescence is strong enough to detect the presence of miR-21. The intensity change follows the concentration dependence of the target miR present in a sample, which provides the basis of developing a new, simple probe for miR detection. The detection strategy is specific, as demonstrated using the response of the DNA/AgNC probe towards the scrambled miR-21 sequence and miR-25 molecule. Additionally, the design reported here is very sensitive with an estimated detection limit at ~1 picomole of miR-21.
Project description:We observed silver nanocolloids (SNCs) exposure to medaka embryo induced morphological deformities like shortened body and undeveloped head and eyes, and our gene expression analysis using medaka oligo DNA microarray suggested that glycan/glycosylation genes, such as gns and alg2, might be the targets of SNCs' toxicity. Validation studies of gns and alg2 gene expressions by qPCR showed stage-dependent and altered mRNA expression patterns, and their altered expressions were more severe in earlier stages of embryogenesis. A result from structural analysis of glycan revealed that stage 11 was the most sensitive stage to SNCs exposure, and stress seemed to be imposed upon endplasmic reticulum (ER) by SNCs. In fact, qPCR analysis of ER mannosidase demonstrated that N-glycan synthesis in ER was inhibited by SNCs. Overall, this study revealed that SNCs exposure caused embryonic malformations through disruption of glycan biosynthetic pathway in ER.
Project description:All over the world, one of the major challenges is the green synthesis of potential materials against antimicrobial resistance and viruses. This study demonstrates a simple method like chemistry lab titration to synthesize green, facile, scalable, reproducible, and stable synergistic silver chloride/benzyldimethylhexadecyl-ammonium chloride (AgCl/BAC) colloidal Nanoantimicrobials (NAMs). Nanocolloidal dispersions of AgCl in an aqueous medium are prepared by using silver nitrate (AgNO3) as precursor and BAC as both sources of chloride and stabilizer, holding an asymmetric molecular structure. The synthetic approach is scalable and green. Both the morphology and stability of AgCl/BAC nanocolloids (NCs) were investigated as a function of different molar fractions of the reagents. AgCl/BAC NCs were characterized by transmission electron microscopy (TEM) and X-ray photoelectron and UV-vis spectroscopies. Zeta potential measurements revealed increasing positive potential values at every stage of the synthesis. Size distribution and hydrodynamic diameter of the particles were measured by dynamic light scattering (DLS), which predicted the formation of BAC layered structures associated with the AgCl nanoparticles (NPs). Small-angle X-ray scattering (SAXS) experiments verify the thickness of the BAC bilayer around AgCl. The produced AgCl/BAC NCs probably have synergistic antimicrobial properties from the AgCl core and the biocide BAC shell. AgCl/BAC NCs stability over months was investigated. The experimental evidence supports the morphological stability of the AgCl/BAC NCs, while higher positive zeta potential values anticipate a long-term antimicrobial effect: a higher surface charge causes NPs to be potentially more lethal to bacteria. AgCl/BAC antimicrobial aqueous colloidal suspensions will be used as additives for the industrial production of antimicrobial coatings.
Project description:Xanthine is a nucleobase, deriving from adenine and guanine by deamination and oxidation processes, which may deposit in the human body causing diseases, similar to uric acid. Here, we have investigated the adsorption of xanthine on silver colloidal nanoparticles by means of surface-enhanced Raman scattering (SERS) with an exciting radiation in the near-infrared spectral region, where interference due to fluorescence does not occur, along with density functional theory calculations of molecule/metal model systems. By adopting a combined experimental and computational approach, we have identified the "marker" SERS bands of xanthine and the tautomer that preferentially binds the silver particles, as well as the molecular group involved in the interaction with metal. This investigation allows using the FT-SERS spectroscopy for biosensory and diagnostic purposes in body fluids, detecting abnormal levels of xanthine, and preventing metabolic diseases.
Project description:Acid-sensing ion channels (ASICs) are trimeric cation-selective ion channels activated by protons in the physiological range. Recent reports have revealed that postsynaptically localized ASICs contribute to the excitatory postsynaptic current by responding to the transient acidification of the synaptic cleft that accompanies neurotransmission. In response to such brief acidic transients, both recombinant and native ASICs show extremely rapid deactivation in outside-out patches when jumping from a pH 5 stimulus to a single resting pH of 8. Given that the resting pH of the synaptic cleft is highly dynamic and depends on recent synaptic activity, we explored the kinetics of ASIC1a and 1a/2a heteromers to such brief pH transients over a wider [H+] range to approximate neuronal conditions better. Surprisingly, the deactivation of ASICs was steeply dependent on the pH, spanning nearly three orders of magnitude from extremely fast (<1 ms) at pH 8 to very slow (>300 ms) at pH 7. This study provides an example of a ligand-gated ion channel whose deactivation is sensitive to agonist concentrations that do not directly activate the receptor. Kinetic simulations and further mutagenesis provide evidence that ASICs show such steeply agonist-dependent deactivation because of strong cooperativity in proton binding. This capacity to signal across such a large synaptically relevant bandwidth enhances the response to small-amplitude acidifications likely to occur at the cleft and may provide ASICs with the ability to shape activity in response to the recent history of the synapse.
Project description:Conceptual and commercial examples of implantable sensors have been limited to a relatively small number of target analytes, with a strong focus on glucose monitoring. Recently, surface-enhanced Raman spectroscopy (SERS) pH sensors were demonstrated to track acid-producing enzymatic reactions targeting specific analytes. We show here that SERS pH tracking in the basic regime is also possible, and can be used to monitor urea concentration. To accomplish this, we developed a hydrogel consisting of polyelectrolyte multilayer microcapsules containing a SERS-sensitive pH reporter (4-mercapopyridine capped silver nanoparticles modified with bovine serum albumin). This pH sensing material exhibited a sensitive Raman scattering response to a wide range of pH from 6.5-9.7. By incorporating urease into the hydrogel matrix, the new sensor was capable of distinguishing urea concentrations of 0, 0.1, 1, and 10 mM. We also found that bovine serum albumin (BSA) prevented severe aggregation of the nanoparticle-based pH sensor, which improved sensing range and sensitivity. Furthermore, BSA safeguarded the pH sensor during the encapsulation procedure. Together, the combination of materials represents a novel approach to enabling optical sensing of reactions that generate pH changes in the basic range.
Project description:Microsatellite loci were developed for the rare, Himalayan, endemic haploid lichen fungus, Lobaria pindarensis, to study its population subdivision and the species' response to forest disturbance and fragmentation. • We developed 18 polymorphic microsatellite markers using 454 pyrosequencing data and assessed them in 109 individuals. The number of alleles per locus ranged from three to 11 with an average of 6.9. Nei's unbiased gene diversity, averaged over loci, ranged from 0.514 to 0.685 in the three populations studied. The cross-amplification success with related species (L. chinensis, L. gyrophorica, L. isidiophora, L. orientalis, L. pulmonaria, L. spathulata, and Lobaria sp.) was generally high and decreased with decreasing relationship to L. pindarensis. • The new markers will allow the study of genetic diversity and differentiation within L. pindarensis across its distribution. Moreover, they will enable us to study the effects of forest management on the genetic population structure of this tree-colonizing lichen and to carry out population genetic studies of related species in East Asia.
Project description:Surface encapsulation of metal nanoparticles (NPs) is fundamental to achieve sufficient dispersion stability of metal nanocolloids, or metal nanoink. However, the feature is incompatible with surface reactive nature of the metal NPs, although these features are both essential to realizing the functional applications into printed electronics technologies. Here we show that two different kinds of encapsulation for silver NPs (AgNPs) by alkylamine and alkylacid together are the key to achieve unique compatibility between the high dispersion stability as dense nanoclolloids and the AgNP chemisorption printing on activated patterned polymer surfaces. Advanced confocal dynamic light scattering study reveals that an additive trace amount of oleic acid is the critical parameter for controlling the dispersion and coagulative (or surface-reactive) characteristics of the silver nanocolloids. The composition of the disperse media is also important for obtaining highly concentrated but low-viscosity silver nanocolloids that show very stable dispersion. The results demonstrate that the high-resolution AgNP chemisorption printing is possible only by using unique silver nanocolloids composed of an exceptional balance of ligand formulation and dispersant composition.
Project description:Silver nanomaterials have been mainly developed as antibacterial healthcare products worldwide, because of their antibacterial activity. However, there is little data regarding the potential risks and effects of large amounts of silver nanomaterials on plants. In contrast, N-glycans play important roles in various biological phenomena, and their structures and expressions are sensitive to ambient environmental changes. Therefore, to assesse the effects of silver nanomaterials, we focused on the correlation between N-glycans and the effects of silver nanomaterials in plants and analyzed N-glycan structures in Oryza sativa seedlings exposed to silver nanocolloids (SNCs). The phenotype analysis showed that the shoot was not affected by any SNC concentrations, whereas the high SNC exposed root was seriously damaged. Therefore, we performed comparative N-glycan analysis of roots. As a result, five of total N-glycans were significantly increased in SNC exposed roots, of which one was a free-N-glycan with one beta-N-acetylglucosamine residue at the reducing end. Our results suggest that the transition of plant complex type N-glycans, including free-N-glycans, was caused by abnormalities in O. sativa development, and free-N-glycan itself has an important role in plant development. This study originally adapted glycome transition analysis to environmental toxicology and proposed a new category called "Environmental glycobiology".
Project description:Planar and lithography-free metal-insulator-metal (MIM) resonators based on the Fabry-Pérot etalon are attractive for biochemical sensing applications because of their acceptable optical performance and cost-effectiveness. However, injecting analytes into the insulating layer where the optical field is localized (high light-matter interaction) is difficult. Here, planar and lithography-free MIM resonators interacting with their environment are reported. In the MIM, molecules of a liquid can infiltrate the inherent nanopores in the deposited silver nanomembrane and be absorbed into the silk protein hydrogel membrane. The silk layer is swollen when water molecules are absorbed, inducing a large shift in the resonance wavelength. Thus, in this study, the proposed MIM resonator was applied as a highly sensitive water sensor, and a water content as low as 0.008% in organic solvents could be determined by reading the shift in the transmission peak. This limit can be lowered further by using a high-resolution spectrometer and a thicker silk layer. In addition, the area of interaction can be artificially selected by applying an elastomer stamp and a patterned photoresist window.