Accurate determination of the concentration and viability of the viral vaccine vectors is urgently needed for preventing the spread of the viral infections, but also supporting the development and ...assessment of recombinant virus-vectored vaccines. Herein, we describe a nanoplasmonic biosensor with nanoscale robot hand structure (Nano RHB) for the rapid, direct, and specific capture and quantification of adenovirus particles. The nanorobot allows simple operation in practical applications, such as real-time monitoring of vaccine quantity and quality, and evaluation of vaccine viability. Modification of the Nano RHB with branched gold nanostructures allow rapid and efficient assessment of human adenovirus viability, with ultrahigh detection sensitivity of only 100 copies/mL through one-step sandwich method. Nano RHB detection results were consistent with those from the gold standard median tissue culture infectious dose and real-time polymerase chain reaction assays. Additionally, the Nano RHB platform showed high detection specificity for different types of viral vectors and pseudoviruses. Altogether, these results demonstrate that the Nano RHB platform is a promising tool for efficient and ultrasensitive assessment of vaccines and gene delivery vectors.
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Previous studies showed that lanthanum hydroxide (LH) has a therapeutic effect on chronic kidney disease (CKD) and vascular calcification, which suggests that it might have clinical value. However, ...the target and mechanism of action of LH are unclear. Metabolomics of clinical samples can be used to predict the mechanism of drug action. In this study, metabolomic profiles in patients with end-stage renal disease (ESRD) were used to screen related signaling pathways, and we verified the influence of LH on the ROS-PI3K-AKT-mTOR-HIF-1α signaling pathway by western blotting and quantitative real-time RT-qPCR in vivo and in vitro. We found that ROS and SLC16A10 genes were activated in patients with ESRD. The SLC16A10 gene is associated with six significant metabolites (L-cysteine, L-cystine, L-isoleucine, L-arginine, L-aspartic acid, and L-phenylalanine) and the PI3K-AKT signaling pathway. The results showed that LH inhibits the ESRD process and its cardiovascular complications by inhibiting the ROS-PI3K-AKT-mTOR-HIF-1α signaling pathway. Collectively, LH may be a candidate phosphorus binder for the treatment of vascular calcification in ESRD.
Interactions between nucleic acids and proteins are essential to genetic information processing. The detection of size changes in nucleic acids is the key to mapping such interactions, and usually ...requires substrates with fluorescent, electrochemical or radioactive labels. Recently, methods have been developed to tether DNA to highly water-soluble Au nanoparticles, and nanoparticle pairs linked by DNA have been used to measure nanoscale distances. Here we demonstrate a molecular ruler in which double-stranded DNA is attached to a Au nanoparticle. The change in plasmon resonance wavelength of individual Au-DNA conjugates depends on the length of the DNA and can be measured with subnanometre axial resolution. An average wavelength shift of approximately 1.24 nm is observed per DNA base pair. This system allows for a label-free, quantitative, real-time measurement of nuclease activity and also serves as a new DNA footprinting platform, which can accurately detect and map the specific binding of a protein to DNA.
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•Zein films deposited on glass cover slips enhanced NIH3T3 cell spreading and viability.•Cellular response to zein surfaces is believed to involve the polyglutamine peptides in zein ...structure.•Zein is a substrate for transglutaminase where zein is a glutamine donor.•The mechanism for zein enhanced cell spreading may be cross-linking between zein and transglutaminase excreted from 3T3 cells.
Zein is a biocompatible corn protein potentially useful in the development of biomaterials. In this study, the deposition of zein on oxygen plasma treated glass cover slips significantly enhanced cell spreading and viability. The mechanism for cellular response to zein coated surfaces was thought to involve the polyglutamine peptides on the zein structure. We hypothesized that zein was a substrate for tissue transglutaminase (tTG), an extracellular enzyme involved in cell-surface interactions. SDS-PAGE results suggested an interaction between zein and tTG, where zein was the glutamine donor. Cross-linking between zein and tTG may be the first step in successful cell adhesion and spreading.
We present novel gold nanophotonic crescent moon structures with a sub-10 nm sharp edge, which can enhance local electromagnetic field at the edge area. The formation of unconventional nanophotonic ...crescent moon structure is accomplished by using a sacrificial nanosphere template and conventional thin film deposition method, which allows an effective batch nanofabrication and precise controls of nanostructure shapes. Unique multiple scattering peaks are observed in a single gold nanocrescent moon with dark-field white light illumination. A 785 nm near-infrared (NIR) diode laser was used as the excitation source to induce the amplified scattering field on the sharp edge of the single gold nanocrescent moon. The Raman scattering spectrum of Rhodamine 6G molecules adsorbed on the single gold nanocrescent moon are characterized, and the Raman enhancement factor of single gold nanocrescent moon is estimated larger than 1010, which suggests the potential applications of gold nanocrescent moons in ultrasensitive biomolecular detection and cellular imaging using surface enhanced Raman spectroscopy.
African swine fever (ASF) is one of the most serious transnational swine diseases in the world. The case fatality rate of susceptible pigs is up to 100%. Currently, no commercial vaccine is ...available, so the prevention and control of ASF mainly relies on early diagnosis and culling of infected pigs. As the ASF virus continues to evolve, develop, and diversify, nucleic acid testing becomes less efficient. Here, we developed a method for the rapid and direct optical measurement of African swine fever virus (ASFV) antibody
. This one-step procedure requires nearly no sample preparation and involves p30 protein-specific label-free integration into standard 96-well plates. Using a nanoplasmonic biosensor with extraordinary optical transmission (EOT) effect, one-step sample addition, ASFV antibody was detected within 20 min. The positive antibody showed a satisfactory sensitivity and linear relationship in the dilution ratio of 1:100-1:16000. It was used for the detection of clinical serum samples with a coincidence rate of 96.6%. The measurement results can be automatically analyzed and displayed on a conventional microplate meter computer and connected device. Our detection method can be widely applied in point-of-care testing (POCT) of ASFV antibody in pig farms.
African swine fever (ASF) is a serious transnational disease caused by the African swine fever virus (ASFV), which is highly contagious in wild boars and domestic pigs. There is currently no available vaccine for ASF; therefore, development efforts are a key priority as ASFV continues to evolve and diversify. The ASF antibody rapid detection platform comprising the nanoplasmonic biosensor with extraordinary optical transmission effect can greatly reduce the detection time and improve detection flux while maintaining detection sensitivity and specificity. The one-step sample addition can effectively avoid cross contamination of samples in the detection process. The detection method provides a solution for the rapid and accurate real-time monitoring of ASF in pig farms.
Understanding the characteristics of rice productivity is of great importance for achieving high yield formation. However, such traits have not yet been studied for different ages of hydroponically ...grown long-mat rice seedlings (HLMS), which constitutes a new method of seedling cultivation. Field experiments were conducted to evaluate the effects of seedling age on the growth stage, photosynthesis characteristics, dry matter production, and yield of HLMS. A conventional japonica rice cultivar (Wuyunjing 24) and an indica hybrid rice cultivar (6 Liangyou 9368) were used as test materials. The results showed that the whole phase was shortened by 13–15 days for young seedlings (13-day-old) compared with old seedlings (27-day-old), which occurred because the growth process accelerated with the transplantation of young seedlings. As seedling age increased, the dry matter weight of stems of individual plants and of the population increased at the transplanting stage but decreased at the maturity stage (MS). Compared with that of 27-day-old seedlings, the average ratio of panicle weight to total plant dry weight of 13-day-old seedlings during a 2-year period increased by 3.71% for Wuyunjing 24 and by 3.78% for 6 Liangyou 9368 at the MS. Moreover, as seedling age increased, the leaf area index and photosynthetic potential decreased for both cultivars, and the photosynthetic rate markedly decreased at the heading stage (HS). With the exception of that of Wuyunjing 24 from the jointing stage to the HS in 2014, the crop growth rate was higher for young seedlings than for old seedlings. Grain yield significantly decreased with seedling age, but no significant difference was detected between the 13- and 20-day-old seedlings for either cultivar. Therefore, equilibrious and high biological yield formation, vigorous growth in the late stages, and high photosynthetic production capacity are important characteristics and causes of the efficient and sustainable output of photosynthetic systems and for achieving high yield formation in young transplanted seedlings (13–20-day-old).
Lodging is an important factor limiting rice yield and quality by bending or breaking stem in japonica rice（Oryza sativa L.） production. The objectives of this study were to determine the mechanism ...of lodging resistance in japonica rice as affected by carbohydrate components, especially its related arrangement in culm tissue and response to top-dressing nitrogen（N） fertilizer. Field experiments were conducted in Danyang County, Jiangsu Province, China, by using two japonica rice varieties Wuyunjing 23（lodging-resistance variety） and W3668（lodging-susceptible variety） with three top-dressing N fertilizer rates（0, 135 and 270 kg N ha~（-1）） in 2013 and 2014. Lodging related physical parameters, morphological characteristics and stem carbohydrate components were investigated at 30 d after full heading stage. Results showed that with increasing N fertilizer rates, the lodging rate and lodging index increased rapidly primarily due to significant reduction of breaking strength in two japonica rice varieties. Correlation analysis revealed that breaking strength was significantly and positively correlated with bending stress, but negatively correlated with section modulus, except for significant correlation at W3668 in 2014. Higher stem plumpness status and structural carbohydrate contents significantly enhanced stem stiffness, despite of lower non-structural carbohydrate. With higher N fertilizer rate, the culm wall thickness was almost identical, and culm diameter increased slightly. The structural carbohydrates, especially for lignin content in culm, reduced significantly under high N rate. Further histochemical staining analysis revealed that high N treatments decreased the lignin deposition rapidly in the sclerenchyma cells of mechanical tissue, large vascular bundle and small vascular bundle region, which were consistent with reduction of bending stress, especially for W3668 and thus, resulted in poor stem strength and higher lodging index. These results suggested that structural carbohydrate plays a vital role for improving stem strength in japonica rice. N rate decreased lodging resistance primarily due to poor stem stiffness, by reducing structural carbohydrate content and lignin deposition in the secondary cell wall of lower internode culm tissue.
Detection and characterization of tumor cells are important for cancer diagnosis and therapy. Various surface attachment methods have been proposed for the capture and enumeration of cancer cells, ...including immunoaffinity and nanostructured surfaces. Zein, a corn protein, has shown good biocompatibility with human liver cells (HL-7702) and mice fibroblasts (NIH3T3). In previous work, it was found that tissue transglutaminase coated on zein substrates enhanced adhesion and spreading of NIH3T3 fibroblast cells. In the present study, cancer cell adhesion to nanostructured zein substrates was investigated. MDA-MB-231 and HeLa cells were used as cancer cell surrogates. MDA-MB-231 cell immobilization was enhanced on zein films, prepared with 80% ethanol, over glass surfaces. The application of tissue transglutaminase onto nanostructured zein substrates further increased cell spreading and adhesion. Cell immobilization increased linearly with the tissue transglutaminase content of the substrate. The effect of substrate morphology was also investigated by seeding cells on electrospun zein fibers. HeLa cell adhesion was also enhanced by zein substrates. This study provided preliminary supporting evidence for developing a zein platform for circulating tumor cells’ immobilization.
Cotton plants are recalcitrant with regards to transformation and induced regeneration. In the present study, 5-enolpyruvylshikimate-3-phosphate (EPSPS), a glyphosate resistant gene from the ...bacterium Agrobacterium sp. strain CP4, was introduced into an elite Bt transgenic cotton cultivar with a modified technique involving in planta Agrobacteriummediated transformation of shoot apex. Primary transformants were initially screened using a 0.26％ glyphosate spray and subsequently by PCR analysis. Five out of 4000 transformants from T1 seeds were obtained resulting in an in planta transformation rate of 0.125％. Four homozygous lines were produced by continuous self-fertilization and both PCR-based selection and glyphosate resistance. Transgene insertion was analyzed by Southern blot analysis. Gene transcription and protein expression levels in the transgenic cotton lines were further investigated by RT-PCR, Western blot, and ELISA methods. Transgenic T3 plants were resistant to as much as 0.4％ of glyphosate treatments in field trials. Our results indicate that the cotton shoot apex transformation technique which is both tissue-culture and genotype-independent would enable the exploitation of transgene technology in different cotton cultivars. Since this method does not require sterile conditions, the use of specialized growth media or the application of plant hormones, it can be conducted under the greenhouse condition.
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