Graphene Oxide-Upconversion Nanoparticle Based Portable Sensors for Assessing Nutritional Deficiencies in Crops.

The development of innovative technologies to rapidly detect biomarkers associated with nutritional deficiencies in crops is highly relevant to agriculture and thus could impact the future of food security. Zinc (Zn) is an important micronutrient in plants, and deficiency leads to poor health, quality, and yield of crops. We have developed portable sensors, based on graphene oxide and upconversion nanoparticles, which could be used in the early detection of Zn deficiency in crops, sensing mRNAs encoding members of the ZIP-transporter family in crops. ZIPs are membrane transport proteins, some of which are up-regulated at the early stages of Zn deficiency, and they are part of the biological mechanism by which crops respond to nutritional deficiency. The principle of these sensors is based on the intensity of the optical output resulting from the interaction of oligonucleotide-coated upconversion nanoparticles and graphene oxide in the absence or presence of a specific oligonucleotide target. The sensors can reliably detect mRNAs in RNA extracts from plants using a smartphone camera. Our work introduces the development of accurate and highly sensitive sensors for use in the field to determine crop nutrient status and ultimately facilitate economically important nutrient management decisions.

Multiplexed mRNA Sensing and Combinatorial-Targeted Drug Delivery Using DNA-Gold Nanoparticle Dimers.

The design of nanoparticulate systems which can perform multiple synergistic functions in cells with high specificity and selectivity is of great importance in applications. Here we combine recent advances in DNA-gold nanoparticle self-assembly and sensing to develop gold nanoparticle dimers that are able to perform multiplexed synergistic functions within a cellular environment. These dimers can sense two mRNA targets and simultaneously or independently deliver one or two DNA-intercalating anticancer drugs (doxorubicin and mitoxantrone) in live cells. Our study focuses on the design of sophisticated nanoparticle assemblies with multiple and synergistic functions that have the potential to advance sensing and drug delivery in cells.

Different chemical strategies to aminate oxidised multi-walled carbon nanotubes for siRNA complexation and delivery.

In this work, we have investigated the preparation of amino-functionalised multi-walled carbon nanotubes (MWCNTs) as potential carriers for the delivery of siRNA. Several studies have shown promising results exploiting functionalised CNTs for the delivery of genetic material in vitro and in vivo. Our groups have previously observed that the type of surface functionalisation used to modify oxidised MWCNTs (oxMWCNTs) can lead to significant differences in nanotube cellular uptake and delivery capability. In those studies, amino-functionalised CNTs were obtained by cycloaddition reactions. Here, we focused on the direct conversion of the carboxylic groups present on oxMWCNTs into amines, and we attempted different synthetic strategies in order to directly tether the amines onto the CNTs, without extending the lateral chain. The functionalised material was characterised by X-ray photoelectron spectroscopy, Fourier transform infra-red spectroscopy and transmission electron microscopy, and the most water-dispersible CNTs were selected for siRNA complexation and cellular uptake studies. The aminated conjugates are demonstrated to be promising vectors to achieve intracellular transport of genetic information.

[60]Fullerene derivative modulates adenosine and metabotropic glutamate receptors gene expression: a possible protective effect against hypoxia.

BACKGROUND: Glutamate, the main excitatory neurotransmitter, is involved in learning and memory processes but at higher concentration results excitotoxic causing degeneration and neuronal death. Adenosine is a nucleoside that exhibit neuroprotective effects by modulating of glutamate release. Hypoxic and related oxidative conditions, in which adenosine and metabotropic glutamate receptors are involved, have been demonstrated to contribute to neurodegenerative processes occurring in certain human pathologies. RESULTS: Human neuroblastoma cells (SH-SY5Y) were used to evaluate the long time (24, 48 and 72 hours) effects of a [60]fullerene hydrosoluble derivative (t3ss) as potential inhibitor of hypoxic insult. Low oxygen concentration (5% O2) caused cell death, which was avoided by t3ss exposure in a concentration dependent manner. In addition, gene expression analysis by real time PCR of adenosine A1, A2A and A2B and metabotropic glutamate 1 and 5 receptors revealed that t3ss significantly increased A1 and mGlu1 expression in hypoxic conditions. Moreover, t3ss prevented the hypoxia-induced increase in A2A mRNA expression. CONCLUSIONS: As t3ss causes overexpression of adenosine A1 and metabotropic glutamate receptors which have been shown to be neuroprotective, our results point to a radical scavenger protective effect of t3ss through the enhancement of these neuroprotective receptors expression. Therefore, the utility of these nanoparticles as therapeutic target to avoid degeneration and cell death of neurodegenerative diseases is suggested.

Modulation of gene expression of adenosine and metabotropic glutamate receptors in rat's neuronal cells exposed to L-glutamate and [60]fullerene.

L-Glutamate (L-Glu) has been often associated not only to fundamental physiological roles, as learning and memory, but also to neuronal cell death and the genesis and development of important neurodegenerative diseases. Herein we studied the variation in the adenosine and metabotropic glutamate receptors expression induced by L-Glu treatment in rat's cortical neurons. The possibility to have structural alteration of the cells induced by L-Glu (100 nM, 1 and 10 microM) has been addressed, studying the modulation of microtubule associated protein-2 (MAP-2) and neurofilament heavy polypeptide (NEFH), natively associated proteins to the dendritic shape maintenance. Results showed that the proposed treatments were not destabilizing the cells, so the L-Glu concentrations were acceptable to investigate fluctuation in receptors expression, which were studied by RT-PCR. Interestingly, C60 fullerene derivative t3ss elicited a protective effect against glutamate toxicity, as demonstrated by MTT assay. In addition, t3ss compound exerted a different effect on the adenosine and metabotropic glutamate receptors analyzed. Interestingly, A(2A) and mGlu1 mRNAs were significantly decreased in conditions were t3ss neuroprotected cortical neurons from L-Glu toxicity. In summary, t3ss protects neurons from glutamate toxicity in a process that appears to be associated with the modulation of the gene expression of adenosine and metabotropic glutamate receptors.

[60]Fullerene-based monolayers as neuroprotective biocompatible hybrid materials.

Here we report on the surface immobilization of redox-active [60]fullerene derivatives and the consequent neuroprotective effects toward l-glutamate induced excitotoxicity in human derived undifferentiated neuroblastoma cells.

Modulation of adenosine receptors by [60]fullerene hydrosoluble derivative in SK-N-MC cells.

The most known fullerenes are spherical carbon compounds composed of 60 carbon atoms. C(60) fullerenes have shown biochemical and biomedical properties in the last years such as as blockade of apoptosis and neuroprotection. The nucleoside adenosine has a neuroprotective role mainly due to inhibition of glutamate release, which is a neurotransmitter related to excitotoxicity and cell death. In the present work, we have determined the presence of adenosine receptors in SK-N-MC cells, a neuroepithelioma human cell line, and analyzed the effect of fullerenes in these receptors by using radioligand binding, immunoblotting, and quantitative real time PCR assays. Results demonstrated that SK-N-MC cells endogenously express adenosine receptors. Fullerene exposure of these cells did not affect cell viability measured by MTT reduction assay. However, adenosine A(1) and A(2A) receptors were both increased in SK-N-MC cells after treatment. These results suggest for the first time the modulation of adenosine receptors after C(60) fullerenes exposure.