Purified fluorescent nanohybrids based on quantum dot-HER2-antibody for breast tumor target imaging.

Quantum dots (QDs) have been widely used for bioimaging in vivo because of their excellent optical properties. As part of the preparation process of QD-based nanohybrids, purification is an important step for minimizing contaminants and improving the quality of the product. In this work, we describe high-performance size exclusion chromatography (HPSEC) used to purify nanohybrids of CdSe/ZnS QDs and anti-human epidermal growth factor receptor 2 antibodies (QD-HER2-Ab). The unbound antibody and suspended agglomerates were removed from freshly prepared QD-HER2-Ab via HPSEC. Pure and homogeneous QD-HER2-Ab were then used as immunofluorescence target imaging bioprobes in vivo. The QD-HER2-Ab did not cause any obvious acute toxicity in mice one week after a single intravenous injection of 15 nmol/kg. The purified QD-HER2-Ab bioprobes showed high tumor targeting ability in a human breast tumor xenograft nude mouse model (24 h after injected) with the possibility of in vivo immunofluorescence tumor imaging. The immunofluorescence imaging background signal and acute toxicity in vivo were minimized because of the reduction of residual QDs. HPSEC-purified QD-HER2-Ab is an accurate and convenient tool for in vivo tumor target imaging and HER2 detection, thus providing a basis for the purification of other QD-based bioprobes.

Effects of Chitosan Molecular Weight and Degree of Deacetylation on Chitosan-Cellulose Nanocrystal Complexes and Their Formation.

This study was conducted to determine the effects of chitosan molecular weight and degree of deacetylation (DD) on chitosan-cellulose nanocrystal (CNC) polyelectrolyte-macroion complexes (PMCs) and their formation. Chitosan samples with three different molecular weights (81, 3 · 103, 6 · 103 kDa) and four different DDs (77, 80, 85, 89%) were used. The effects on PMC formation were determined by turbidimetric titration. An effect of the molecular weight of chitosan was not observed in turbidimetric titrations. Turbidity levels were higher for CNCs with lower sulfate group density and larger hydrodynamic diameter than for CNCs with higher sulfate group density and smaller hydrodynamic diameter. Conversely, turbidity levels were higher for chitosans with higher DD (higher charge density) than for chitosans with lower DD (lower charge density). PMC particles from chitosans with different molecular weights were characterized by scanning electron microscopy, laser Doppler electrophoresis, and dynamic light scattering. PMCs from high-molecular-weight chitosan were more spherical and those from medium-molecular-weight chitosan had a slightly larger hydrodynamic diameter than PMCs from the respective other two chitosans. The molecular weight of the chitosan was concluded to have no effect on the formation of chitosan-CNC PMC particles and only a minor effect on the shape and size of the particles. The higher turbidity levels for CNCs with lower sulfate group density and larger hydrodynamic diameter and for chitosans with higher DD were attributed to a larger number of CNCs being required for charge compensation.

Mode of action of nanochitin whisker against Fusarium pseudograminearum.

Nanochitin whisker (NC) is an advanced nanobiomaterial with novel physicochemical and biological properties. Fusarium pseudograminearum (Fpg) is an important pathogenic fungus causing wheat crown rot disease. This study explored the mode of action of NC against Fpg as a target microorganism. The effects of different treatments and concentrations of NC on the fungal growth and conidial germination were investigated by in vitro bioassay. The impacts of NC on cell structure integrity, membrane permeability, pathogenesis related key enzymes activity, and the mycotoxin production were examined by electron microscopy, fluorescence spectroscopy, IR spectroscopy, conductometry, and spectrophotometry, respectively. The results showed that NC significantly reduced hyphal growth, and the spore germination rate of Fpg declined by 33.0 % and 23.2 % when Fpg was treated with 30 and 300 µg/mL of NC, respectively. NC vigorously influenced structural stability of cell wall by destroying dextran structure, and strongly stimulated ergosterol production altering membrane integrity of the fungus. It reduced the activities of enzymes related to energy-supply like nicotinamide adenine dinucleotide oxidase and succinate dehydrogenase remarkably. The production of fungal mycotoxin deoxynivalenol was also decreased by NC. These findings provide an important basis for fully understanding the mechanism of nanobiomaterial in plant fungal disease control.

Quaternary ammonium iminofullerenes improve root growth of oxidative-stress maize through ASA-GSH cycle modulating redox homeostasis of roots and ROS-mediated root-hair elongation.

BACKGROUND: Various environmental factors are capable of oxidative stress to result in limiting plant development and agricultural production. Fullerene-based carbon nanomaterials can enable radical scavenging and positively regulate plant growth. Even so, to date, our knowledge about the mechanism of fullerene-based carbon nanomaterials on plant growth and response to oxidative stress is still unclear. RESULTS: 20 or 50 mg/L quaternary ammonium iminofullerenes (IFQA) rescued the reduction in root lengths and root-hair densities and lengths of Arabidopsis and maize induced by accumulation of endogenous hydrogen peroxide (H2O2) under 3-amino-1,2,4-triazole or exogenous H2O2 treatment, as well as the root active absorption area and root activity under exogenous H2O2 treatment. Meanwhile, the downregulated contents of ascorbate acid (ASA) and glutathione (GSH) and the upregulated contents of dehydroascorbic acid (DHA), oxidized glutathione (GSSG), malondialdehyde (MDA), and H2O2 indicated that the exogenous H2O2 treatment induced oxidative stress of maize. Nonetheless, application of IFQA can increase the ratios of ASA/DHA and GSH/GSSG, as well as the activities of glutathione reductase, and ascorbate peroxidase, and decrease the contents of H2O2 and MDA. Moreover, the root lengths were inhibited by buthionine sulfoximine, a specific inhibitor of GSH biosynthesis, and subsequently rescued after addition of IFQA. The results suggested that IFQA could alleviate exogenous-H2O2-induced oxidative stress on maize by regulating the ASA-GSH cycle. Furthermore, IFQA reduced the excess accumulation of ROS in root hairs, as well as the NADPH oxidase activity under H2O2 treatment. The transcript levels of genes affecting ROS-mediated root-hair development, such as RBOH B, RBOH C, PFT1, and PRX59, were significantly induced by H2O2 treatment and then decreased after addition of IFQA. CONCLUSION: The positive effect of fullerene-based carbon nanomaterials on maize-root-hair growth under the induced oxidative stress was discovered. Application IFQA can ameliorate oxidative stress to promote maize-root growth through decreasing NADPH-oxidase activity, improving the scavenging of ROS by ASA-GSH cycle, and regulating the expressions of genes affecting maize-root-hair development. It will enrich more understanding the actual mechanism of fullerene-based nanoelicitors responsible for plant growth promotion and protection from oxidative stress.

Quaternary ammonium iminofullerenes promote root growth and osmotic-stress tolerance in maize via ROS neutralization and improved energy status.

In the present study, the role of quaternary ammonium iminofullerenes (IFQA) on the root growth of plant seedlings was investigated. The root elongation of Arabidopsis and maize exposed to 20 and 50 mg/L of IFQA was promoted under normal and osmotic stress conditions, respectively. In the meantime, the root active absorption area and adenosine triphosphate content in roots of maize seedlings were enhanced by IFQA treatment, however, the contents of hydrogen peroxide (H2O2) and malondialdehyde in roots were down-regulated. IFQA application improved glutathione transferase and glutathione reductase activities and the ratios of glutathione/oxidized glutathione and ascorbic acid/dehydroascorbic acid, and restored the inhibition of root elongation caused by the excess accumulation of H2O2 in roots of maize seedlings under osmotic stress. Furthermore, the expression of 14 proteins involved in cell growth, energy metabolism, and stress response in maize roots was upregulated by two-dimensional electrophoresis combined with mass spectrometry. This analysis revealed that IFQA stimulated the redox pathway to maintain balance levels of reactive oxygen species to ensure normal cell metabolism, promote energy production for root growth, and enhance osmotic-stress tolerance. It provided crucial information to elucidate the mechanism of the root growth of crop seedlings enhanced by water-soluble fullerene-based nanomaterials.

Nanochitin whisker enhances insecticidal activity of chemical pesticide for pest insect control and toxicity.

BACKGROUND: Nanomaterials in plant protection promise many benefits over conventional pesticide products. Nano-enabled pesticides may alter the functionality or risk profile of active ingredients. Cationic nanochitin whiskers (NC) possess strong biological activity against wheat aphids. However, toxicity and synergistic effects of NC with chemical pesticides against pest insects has not been systemically reported. This study investigated the insecticidal enhancement by NC with Omethoate (40% EC), Imidacloprid (10% WP), and Acetamiprid (40% WG) for pest control using wheat aphid as piercing-sucking mouthparts insect. Fluorescein isothiocyanate labelled NC was used to monitor the uptake and transportation pathway of NC inside the target insects. Toxicity of NC was tested with Sprague-Dawley (SD) rat. Our findings provide a theoretical basis for future application of NC in plant protection against pest insects. RESULTS: NCs synthesized by acidic hydrolysis were rod-like nanoparticles in a range of 50-150 nm in length and 30-50 nm in width, which examined by electron microscopy and dynamic light scattering methods. The charge density and zeta potential were about 63 mmol/kg and + 36.4 mV, respectively. By absorption and/or contact action of 30-50 mg/L of NC suspension, the corrected mortality of wheat aphids reached up to 80% or above after 12 h treatment, NC could be distributed through digestive system and relocated from mouth to other tissues inside the insect body. When associated with dilutions of conventional pesticides, the corrected mortality were significantly increased up to 95% or above. The dosage of the chemical pesticide and nanochitin in the mixtures (1:1 by volume) were all reduced to half. The acute oral toxicity Lethal Dose 50% (LD50) to SD rat is greater than 5000 mg/kg BW (body weight) in male and female, acute dermal toxicity LD50 is greater than 2000 mg/kg BW of NC. CONCLUSIONS: NC has a strong promotive effect on insecticidal effectiveness of chemical insecticides. It was easily absorbed by plant, transported and distributed from mouth to other tissues of the insects while sucking plant fluid. Low acute oral and dermal toxicity to SD rat indicated that it is safe to apply in agriculture and food industry. NCs has a great potential for water-based nanopesticide formulation to reduce chemical pesticide use for future agro-environmental sustainability.

Molecular mechanism of nanochitin whisker elicits plant resistance against Phytophthora and the receptors in plants.

Rod-like nanochitin (NC) whisker with cationic nature has a strong synergistic effect with fungicides on inhibition of tobacco root rot disease. This study we explored the activity of NC against Phytophthora and the mechanism for eliciting plant defense response and the receptors in planta. P. capsici isolates, model Nicotiana benthamiana plants and Arabidopsis thaliana were treated with 0.005% of NC suspension and 1 µM of flg22. Infection control efficacy against P. capsici isolates, biosynthetic enzyme activities and the PR genes expression were determined at different hours post treatment in plant. The infection control efficacy, ROS generation, and PTI maker gene expression were re-analyzed in A. thaliana Col-0, bak1 and cerk1 mutants. The results showed that NC did not exhibit inhibitory effect on vegetative growth of P. capsici, but enhanced the resistance against P. capsici by systemically enhanced phenylalanine ammonia-lyase activity and PR gene expression. P. capsici resistance, PTI maker gene promotion, and ROS production in A. thaliana induced by NC depended not only on chitin receptor CERK1, but also BAK1. NC and flg22 induced oomycete immunity through a mechanism of a cross-microbe protection via the BAK1-CERK1 pathway in plant, pointing to the complexity of the plant immunity system.

Quaternary Ammonium Salts of Iminofullerenes: Fabrication and Effect on Seed Germination.

In this study, novel water-soluble quaternary ammonium salts of iminofullerenes (IFQA) were synthesized by nitrene chemistry in combination with quaternization and identified as [C60(NCH2CH2NH3+·CF3COO-)4·10H2O]n by various spectroscopies. Maize and Arabidopsis seeds were used to test the bioactivity of IFQA in seed germination. Compared with the control, maize seed exposure to 50 mg/L IFQA (normal: 73.1% vs 58.7%; drought: 66.7% vs 50.0% at the second day) and Arabidopsis seed exposure to 20 mg/L IFQA (normal: 77.5% vs 58.8%; drought: 63.3% vs 36.7% at the second day) had higher germination rates and quicker germination. The results of two-dimensional gel electrophoresis combined with mass spectroscopy showed that the abundance of 21 proteins in embryo proteome of maize seeds was significantly changed (>1.5 fold). The downregulated six storage proteins and upregulated four proteins induced by IFQA for energy production and sugar metabolism indicated a faster metabolic activity of maize seed germination. The upregulated eight stress-related proteins and antioxidant enzymes suggested that the role of IFQA was to activate the metabolic processes in seed germination and also increase seed stress response. The results provide important information to understand the mechanism of seed germination enhancement by carbon nanomaterials.

The Stimulatory Effects of Nanochitin Whisker on Carbon and Nitrogen Metabolism and on the Enhancement of Grain Yield and Crude Protein of Winter Wheat.

Nanochitin whisker (NC) with a cationic nature could enhance plant photosynthesis, grain yield, and quality of wheat, but have not been systematically studied. This study was designed to investigate the stimulatory effects of NC on dry matter (DM) and nitrogen (N) accumulation and translocation, and on the metabolism of carbon (C) and N in later growth stages of winter wheat to reveal the enhancement mechanism of grain yield and crude protein concentration. Different parts of NC-treated plants from pot grown experiments were collected at the pre- and post-anthesis stages. The accumulation, translocation, and contributions of DM and N from pre-anthesis vegetation organs to grains, as well as key metabolic enzyme activities, including sucrose phosphate synthase (SPS) and phosphoenolpyruvate carboxylase (PEPC), were examined. The results showed that, at an application rate of 6 mg·kg-1 of NC in the soil, the accumulation of DM and N were significantly enhanced by 16.2% and 38.8% in pre-anthesis, and by 15.4% and 30.0% in post-anthesis, respectively. Translocation of N and DM in the post-anthesis periods were enhanced by 38.4% and 50.9%, respectively. NC could also stimulate enzyme activities, and increased 39.8% and 57.1% in flag leaves, and by 36.0% and 58.8% in spikes, respectively, at anthesis. SPS and PEPC increased by 28.2% and 45.1% in flag leaves, and by 42.2% and 56.5% in spikes, respectively, at 15 days after anthesis. The results indicated that the NC promoted N metabolism more than C metabolism, and resulted in the enhancement of grain yield by 27.56% and of crude protein concentration in grain by 13.26%, respectively.

Antifungal Activity of Nanochitin Whisker against Crown Rot Diseases of Wheat.

Nanochitin whisker (NC) is a rodlike particle with a cationic nature and high biological activities. Crown rot, caused by soil-borne fungal pathogens including Fusarium pseudograminearum( Fp) and Fusarium graminearum( Fg), is one of the most damaging diseases in wheat. To explore the antifungal activity of NC against wheat diseases, the effects of nanochitin whisker suspension (NCs) on fungal growth and conidial production of Fp and Fg were studied in vitro. NCs and its mixture with chemical fungicide as a seeds coating agent for crown rot disease control were also investigated using Fp as a model microorganism in a pot test. The results showed that NCs had significant inhibitory effects on mycelial growth and conidial production of Fp and Fg at concentrations of 30 and 300 ppm in the growth medium. Particularly, 300 ppm of NCs was capable of reducing conidial formation 89.25% and 82.28% for Fp and Fg, respectively. When seeds were treated with a mixture of NCs and tebuconazole, the disease control efficiencies increased to 79.30% and 90.02% for NCs at concentrations of 10 and 30 ppm, respectively. Greener and shorter seedlings were also observed in the pot experiment. This indicates that NCs have strong antifungal activity against the soil-borne pathogens of wheat and reduce use of chemical fungicide in wheat plantation.

Effects of Nanochitin on the Enhancement of the Grain Yield and Quality of Winter Wheat.

This study investigated the effects of different rates of nanochitin in soil on the grain yield and quality of winter wheat. Nanochitin obtained by acidic hydrolysis of shrimp chitin was a rod-like whisker possessing a hydrodynamic diameter of 143 nm and ζ potential of 55.7 mV. Two varieties of winter wheat, multi-spike wheat (MSW) and large spike wheat (LSW), were treated with the nanochitin suspension in outside pot experiments. The results showed that 0.006 g kg-1 of nanochitin in soil could significantly enhance the yield by 23.0% for MSW and 33.4% for LSW, with significant increases of net photosynthesis rate, stomatal conductance, intercellular CO2 concentrations, and transpiration rate in flag leaf at the grain filling stage. Grain protein, iron, and zinc contents in wheat treated with nanochitin were also increased by 5.0, 10.3, and 22.1% for MSW and 33.4, 32.0, and 27.0% for LSW, respectively. This indicated that utilization of nanochitin has a great potential in future agriculture sustainability and crop production.

Synergistic effects of nanochitin on inhibition of tobacco root rot disease.

Nanomaterials have great potential for use in various fields, due to their unique properties. In order to explore the bioactivity of nanochitin on tobacco, the effects of nanochitin suspensions on tobacco seed germination, seedling growth, and synergistic effects with fungicides were studied in indoor and field trials. Results showed that 0.004% (w/v) of nanochitin improved tobacco seed germination and shortened mean time to germination significantly; 0.005% (w/v) of nanochitin increased tobacco stem length, stem girth, leaf number and leaf area, and 0.001% (w/v) of nanochitin had synergistic effects on inhibition of tobacco root rot when mixed with metalaxyl mancozeb and thiophanate methyl fungicides. This indicates that nanochitin suspensions have a strong potential to protect tobacco from tobacco root rot diseases and reduce the use of chemical fungicides in tobacco plantations.

Femoral neck fractures: prognosis based on a new classification after superselective angiography.

BACKGROUND: Vascular injury after femoral neck fracture can determine its prognosis. This study aimed to determine methods of evaluating femoral neck fracture and accurately predicting the prognosis. METHODS: Forty-five patients with a single femoral neck fracture all underwent digital subtraction angiography (DSA) and were divided into three types by DSA. DSA showing three to six supporting band vascular images crossing the fracture line was classified as type I. DSA showing one to two supporting band vascular images crossing the fracture line was classified as type II. When DSA did not show vascular images crossing the fracture line, it was classified as type III. The 45 patients were divided according to age into elderly, middle-aged and youth groups. All cases were given internal fixation operations by a hollow screw under a C-brachial machine. After a follow-up of 6-60 months, avascular necrosis of the femoral head (ANFH) and fracture healing of different type and different ages of patients were evaluated according to symptoms and imaging examinations. RESULTS: (1) For DSA types I, II and III, the rates of ANFH were 0, 7.14 and 100 %, respectively, and the rates of fracture disunion were 13.3, 7.15 and 0 %, respectively. Therefore, the rate of ANFH is negatively related to the visible supporting band vascular amount on DSA. (2) In the young group, the proportions of types I, II and III were 6.7, 26.67 and 66.7 %, respectively, 18.18, 36.36 and 45.45 % in the middle-aged group and 63.16, 31.58 and 5.27 % in the elderly group. The rates of necrosis for elderly, middle-aged and youth were 10.53, 45.45 and 66.67 %, respectively, and the rates of fracture disunion were 0, 0 and 6.67 %, respectively. So we can draw the conclusion that the amount of supporting band vascular images is inversely proportional to age and the union is directly related to age, but independent of the supporting band vascular amount of DSA showing. CONCLUSION: The new classification after superselective angiography is valuable for predicting the prognosis of femoral neck fractures.

Quantitative genetic background of the host influences gut microbiomes in chickens.

Host genotype and gender are among the factors that influence the composition of gut microbiota. We studied the population structure of gut microbiota in two lines of chickens maintained under the same husbandry and dietary regimes. The lines, which originated from a common founder population, had undergone 54 generations of selection for high (HW) or low (LW) 56-day body weight, and now differ by more than 10-fold in body weight at selection age. Of 190 microbiome species, 68 were affected by genotype (line), gender, and genotype by gender interactions. Fifteen of the 68 species belong to Lactobacillus. Species affected by genotype, gender, and the genotype by gender interaction, were 29, 48, and 12, respectively. Species affected by gender were 30 and 17 in the HW and LW lines, respectively. Thus, under a common diet and husbandry host quantitative genotype and gender influenced gut microbiota composite.

Effects of pH and salt concentration on the formation and properties of chitosan-cellulose nanocrystal polyelectrolyte-macroion complexes.

This study examines the effects of pH and salt concentration on the formation and properties of chitosan-cellulose nanocrystal (CNC) polyelectrolyte-macroion complexes (PMCs). The components' pK values, determined by potentiometric titration, were 6.40 for chitosan and 2.46 for the CNCs. The turbidity of PMC particle suspensions was measured as a function of chitosan-CNC ratio, pH, and salt concentration. The maximum turbidity values in titrations of a chitosan solution with a CNC suspension and vice versa occurred at charge ratios of 0.47 ± 0.11 (SO(3)(-)/NH(3)(+)) and 1.16 ± 0.06 (NH(3)(+)/SO(3)(-)), respectively. A pH increase caused a turbidity decrease due to shrinking of the PMC particles upon changes in their components' degrees of ionization. An increase in salt concentration caused a decrease in turbidity due to charge-screening-related shrinking of the PMC particles. The effects of pH and salt concentration on particle size were confirmed by scanning electron microscopy.

Formation and properties of chitosan-cellulose nanocrystal polyelectrolyte-macroion complexes for drug delivery applications.

This study examines a novel polyelectrolyte-macroion complex (PMC) between chitosan, a cationic polysaccharide, and cellulose nanocrystals (CNCs), anionic, cylindrical nanoparticles, for potential applications in drug delivery. CNCs were prepared by H(2)SO(4) hydrolysis of wood pulp. The formation of PMCs was monitored by turbidimetric titration. In titrations of a chitosan solution with a CNC suspension, the turbidity reached a plateau, but it had a maximum and then decreased when the direction of titration was reversed. PMC particles were characterized by Fourier transform infrared spectroscopy, scanning electron microscopy, dynamic light scattering, and laser Doppler electrophoresis. The particles were composed primarily of CNCs and ranged in size from a few hundred nanometers to several micrometers, depending on the cellulose/chitosan ratio. Particles formed at amino/sulfate group molar ratios >1 were nearly spherical in shape and positively charged, whereas particles formed at ratios <1 had well-defined nonspherical shapes and were negatively charged.