Next-generation diagnostic test for dengue virus detection using an ultrafast plasmonic colorimetric RT-PCR strategy.

The current global COVID-19 pandemic once again highlighted the urgent need for a simple, cost-effective, and sensitive diagnostic platform that can be rapidly developed for distribution and easy access in resource-limited areas. Here, we present a simple and low-cost plasmonic photothermal (PPT)-reverse transcription-colorimetric polymerase chain reaction (RTcPCR) for molecular diagnosis of dengue virus (DENV) infection. The assay can be completed within 54 min with an estimated detection limit of 1.6 copies/µL of viral nucleic acid. The analytical sensitivity and specificity of PPT-RTcPCR were comparable to that of the reference RT-qPCR assay. Moreover, the clinical performance of PPT-RTcPCR was evaluated and validated using 158 plasma samples collected from patients suspected of dengue infection. The results showed a diagnostic agreement of 97.5% compared to the reference RT-qPCR and demonstrated a clinical sensitivity and specificity of 97.0% and 100%, respectively. The simplicity and reliability of our PPT-RTcPCR strategy suggest it can provide a foundation for developing a field-deployable diagnostic assay for dengue and other infectious diseases.

Plasmonic Cross-Linking Colorimetric PCR for Simple and Sensitive Nucleic Acid Detection.

Simple, low-cost, and accurate nucleic acid assay platforms hold great promise for point-of-care (POC) pathogen detection, disease surveillance, and control. Plasmonic photothermal polymerase chain reaction (PPT-PCR) is a powerful and efficient nucleic acid amplification technique, but it lacks a simple and convenient analysis method for POC applications. Herein, we propose a novel plasmonic cross-linking colorimetric PCR (PPT-ccPCR) assay by integrating plasmonic magnetic nanoparticle (PMN)-based PPT-PCR with gold nanoparticle (AuNP)-based cross-linking colorimetry. AuNPs form assembled structures with the PMNs in the presence of amplicons and collect in a magnetic field, resulting in color changes to the supernatant. Target DNA with concentrations as low as 5 copies/µL can be visually detected within 40 min. The achieved limit of detection was 1.8 copies/µL based on the absorption signals. This simple and sensitive strategy needs no expensive instrumentation and demonstrates high potential for POC detection while enabling further applications in clinical diagnostics.

Terahertz hybrid optical-plasmonic modes: tunable resonant frequency, narrow linewidth, and strong local field enhancement.

Hybrid optical-plasmonic modes have the characteristics of low loss and small mode volume, which will result in the strong localization and enhancement of electromagnetic field. Such advantages of hybrid optical-plasmonic mode are important for the enhancement of light-matter interactions. Here, terahertz (THz) hybrid modes of Fabry-Perot resonances (FPRs) and spoof surface plasmon polaritons (SSPPs) in the modified Otto scheme are investigated both in theoretical and experimental aspects. The device structure is composed of a metal grating silicon waveguide (MGSW) and a metal slit grating (MSG). The two components are vertically stacked with a variable air gap between them. The THz hybrid modes are originated from the far-field coupling of the FPRs and the SSPP supported by the air gap and the MSG, respectively. By changing the thickness of the air gap, the resonant frequency of the FPR-SSPP modes can be tuned in a frequency range of about 0.1 THz. An anti-crossing behavior between two reflection dips corresponding to the guided-mode resonance in the MGSW and the FPR-SSPP mode is observed, which leads to the narrowing of the reflection dips in the anti-crossing region. Numerical simulations show that at the resonant frequencies of FPR-SSPP mode, there is a huge volume-averaged electromagnetic energy enhancement of about 1600 times in the grooves of the MSG, which is around 8.7 times larger than that induced by the SSPP directly launched by free-space electromagnetic field. The hybrid FPR-SSPP modes can be used to construct THz sensors and detectors with high sensitivity.

A rapid and sensitive fluorescence biosensor based on plasmonic PCR.

Plasmonic PCR utilizing metallic nanoparticles has shown great advantages compared to the commercial thermocycler equipment in terms of cost, size and processing time. However, due to the strong fluorescence quenching, plasmonic nanoparticle-based PCR requires additional post-processing steps such as centrifugation and gel electrophoresis. This process increases the overall diagnostic time, offsetting the benefits of fast thermocycling. Here, we report a rapid and sensitive plasmonic photothermal PCR (PPT-PCR) assay method based on in situ end-point fluorescence detection. By using plasmonic magnetic bi-functional nanoparticles, PPT-PCR involving 30 thermocycles and fluorescence detection following magnetic separation has successfully shown that DNA targets can be detected within 5.5 minutes. The limit of detection (3.3 copies per µL) is comparable with that of the conventional real-time quantitative PCR; however, the assay time is about 5.5 times shorter for the PPT-PCR. The strategy of combining the photothermal effect and magnetic separation into a single particle will open new horizons in the development of fast and sensitive PCR-based biosensors for point-of care testing.

Hypothesis-based food, feed, and environmental safety assessment of GM crops: A case study using maize event DP-202216-6.

Event DP-2Ø2216-6 (referred to as DP202216 maize) was genetically modified to increase and extend the expression of the introduced zmm28 gene relative to endogenous zmm28 gene expression, resulting in plants with enhanced grain yield potential. The zmm28 gene expresses the ZMM28 protein, a MADS-box transcription factor. The safety assessment of DP202216 maize included an assessment of the potential hazard of the ZMM28 protein, as well as an assessment of potential unintended effects of the genetic insertion on agronomics, composition, and nutrition. The history of safe use (HOSU) of the ZMM28 protein was evaluated and a bioinformatics approach was used to compare the deduced amino acid sequence of the ZMM28 protein to databases of known allergens and toxins. Based on HOSU and the bioinformatics assessment, the ZMM28 protein was determined to be unlikely to be either allergenic or toxic to humans. The composition of DP202216 maize forage and grain was comparable to non-modified forage and grain, with no unintended effects on nutrition or food and feed safety. Additionally, feeding studies with broiler chickens and rats demonstrated a low likelihood of unintentional alterations in nutrition and low potential for adverse effects. Furthermore, the agronomics observed for DP202216 maize and non-modified maize were comparable, indicating that the likelihood of increased weediness or invasiveness of DP202216 maize in the environment is low. This comprehensive review serves as a reference for regulatory agencies and decision-makers in countries where authorization of DP202216 maize will be pursued, and for others interested in food, feed, and environmental safety.

Maize BIG GRAIN1 homolog overexpression increases maize grain yield.

The Zea Mays BIG GRAIN 1 HOMOLOG 1 (ZM-BG1H1) was ectopically expressed in maize. Elite commercial hybrid germplasm was yield tested in diverse field environment locations representing commercial models. Yield was measured in 101 tests across all 4 events, 26 locations over 2 years, for an average yield gain of 355 kg/ha (5.65 bu/ac) above control, with 83% tests broadly showing yield gains (range +2272 kg/ha to -1240 kg/ha), with seven tests gaining more than one metric ton per hectare. Plant and ear height were slightly elevated, and ear and tassel flowering time were delayed one day, but ASI was unchanged, and these traits did not correlate to yield gain. ZM-BG1H1 overexpression is associated with increased ear kernel row number and total ear kernel number and mass, but individual kernels trended slightly smaller and less dense. The ZM-BG1H1 protein is detected in the plasma membrane like rice OS-BG1. Five predominant native ZM-BG1H1 alleles exhibit little structural and expression variation compared to the large increased expression conferred by these ectopic alleles.

Overexpression of zmm28 increases maize grain yield in the field.

Increasing maize grain yield has been a major focus of both plant breeding and genetic engineering to meet the global demand for food, feed, and industrial uses. We report that increasing and extending expression of a maize MADS-box transcription factor gene, zmm28, under the control of a moderate-constitutive maize promoter, results in maize plants with increased plant growth, photosynthesis capacity, and nitrogen utilization. Molecular and biochemical characterization of zmm28 transgenic plants demonstrated that their enhanced agronomic traits are associated with elevated plant carbon assimilation, nitrogen utilization, and plant growth. Overall, these positive attributes are associated with a significant increase in grain yield relative to wild-type controls that is consistent across years, environments, and elite germplasm backgrounds.

Evaluation of the History of Safe Use of the Maize ZMM28 Protein.

The ZMM28 protein encoded by the zmm28 gene is endogenous to maize. DP202216 maize was genetically modified to increase and extend expression of the zmm28 gene relative to native zmm28 gene expression, resulting in plants with enhanced grain yield potential. Evaluation of the history of safe use (HOSU) is one component of the safety assessment framework for a newly expressed protein in a GM crop. The deduced amino acid sequence of the introduced ZMM28 protein in DP202216 maize is identical to the ZMM28 protein in nonmodified conventional maize. The ZMM28 protein has also been found in selected varieties of sweet corn kernels, and closely related proteins are found in other commonly consumed food crops. Concentrations of the ZMM28 protein in event DP202216 maize, conventional maize, and sweet corn are reported. This information supports, in part, the evaluation of HOSU, which can be leveraged in the safety assessment of the ZMM28 protein. Additional studies will be considered in the food and feed safety assessment of the DP202216 maize event.

Hollow hematite single crystals deposited with ultra-thin Al2O3 by atom layer deposition for improved photoelectrochemical performance.

Hematite (α-Fe2O3) is a red material with a band gap of about 2.0 eV, which indicates that it can absorb more solar light. It is a promising photocatalyst applied in many fields. In this paper, α-Fe2O3 single crystal hollow hexagonal bipyramids were synthesized by a simple one-pot hydrothermal method. The morphology and structure of the prepared α-Fe2O3 hollow hexagonal bipyramids were studied using scanning electron microscopy (SEM), transmission electron microscopy (TEM), and X-ray diffraction (XRD). The hollow single crystals show a good light absorption and performance in photodegradation of methylene blue (MB). Due to the strategy of depositing ultra-thin layers of Al2O3 by atomic layer deposition (ALD), the photoelectrochemical (PEC) performance of α-Fe2O3 under the simulated solar light irradiation is also improved.

Gene expression biomarkers provide sensitive indicators of in planta nitrogen status in maize.

Over the last several decades, increased agricultural production has been driven by improved agronomic practices and a dramatic increase in the use of nitrogen-containing fertilizers to maximize the yield potential of crops. To reduce input costs and to minimize the potential environmental impacts of nitrogen fertilizer that has been used to optimize yield, an increased understanding of the molecular responses to nitrogen under field conditions is critical for our ability to further improve agricultural sustainability. Using maize (Zea mays) as a model, we have characterized the transcriptional response of plants grown under limiting and sufficient nitrogen conditions and during the recovery of nitrogen-starved plants. We show that a large percentage (approximately 7%) of the maize transcriptome is nitrogen responsive, similar to previous observations in other plant species. Furthermore, we have used statistical approaches to identify a small set of genes whose expression profiles can quantitatively assess the response of plants to varying nitrogen conditions. Using a composite gene expression scoring system, this single set of biomarker genes can accurately assess nitrogen responses independently of genotype, developmental stage, tissue type, or environment, including in plants grown under controlled environments or in the field. Importantly, the biomarker composite expression response is much more rapid and quantitative than phenotypic observations. Consequently, we have successfully used these biomarkers to monitor nitrogen status in real-time assays of field-grown maize plants under typical production conditions. Our results suggest that biomarkers have the potential to be used as agronomic tools to monitor and optimize nitrogen fertilizer usage to help achieve maximal crop yields.

Plant nuclear factor Y (NF-Y) B subunits confer drought tolerance and lead to improved corn yields on water-limited acres.

Commercially improved crop performance under drought conditions has been challenging because of the complexity of the trait and the multitude of factors that influence yield. Here we report the results of a functional genomics approach that identified a transcription factor from the nuclear factor Y (NF-Y) family, AtNF-YB1, which acts through a previously undescribed mechanism to confer improved performance in Arabidopsis under drought conditions. An orthologous maize transcription factor, ZmNF-YB2, is shown to have an equivalent activity. Under water-limited conditions, transgenic maize plants with increased ZmNF-YB2 expression show tolerance to drought based on the responses of a number of stress-related parameters, including chlorophyll content, stomatal conductance, leaf temperature, reduced wilting, and maintenance of photosynthesis. These stress adaptations contribute to a grain yield advantage to maize under water-limited environments. The application of this technology has the potential to significantly impact maize production systems that experience drought.