Development and evaluation of a meat mitochondrial metagenomic (3MG) method for composition determination of meat from fifteen mammalian and avian species.

BACKGROUND: Bioassessment and biomonitoring of meat products are aimed at identifying and quantifying adulterants and contaminants, such as meat from unexpected sources and microbes. Several methods for determining the biological composition of mixed samples have been used, including metabarcoding, metagenomics and mitochondrial metagenomics. In this study, we aimed to develop a method based on next-generation DNA sequencing to estimate samples that might contain meat from 15 mammalian and avian species that are commonly related to meat bioassessment and biomonitoring. RESULTS: In this project, we found the meat composition from 15 species could not be identified with the metabarcoding approach because of the lack of universal primers or insufficient discrimination power. Consequently, we developed and evaluated a meat mitochondrial metagenomics (3MG) method. The 3MG method has four steps: (1) extraction of sequencing reads from mitochondrial genomes (mitogenomes); (2) assembly of mitogenomes; (3) mapping of mitochondrial reads to the assembled mitogenomes; and (4) biomass estimation based on the number of uniquely mapped reads. The method was implemented in a python script called 3MG. The analysis of simulated datasets showed that the method can determine contaminant composition at a proportion of 2% and the relative error was < 5%. To evaluate the performance of 3MG, we constructed and analysed mixed samples derived from 15 animal species in equal mass. Then, we constructed and analysed mixed samples derived from two animal species (pork and chicken) in different ratios. DNAs were extracted and used in constructing 21 libraries for next-generation sequencing. The analysis of the 15 species mix with the method showed the successful identification of 12 of the 15 (80%) animal species tested. The analysis of the mixed samples of the two species revealed correlation coefficients of 0.98 for pork and 0.98 for chicken between the number of uniquely mapped reads and the mass proportion. CONCLUSION: To the best of our knowledge, this study is the first to demonstrate the potential of the non-targeted 3MG method as a tool for accurately estimating biomass in meat mix samples. The method has potential broad applications in meat product safety.

Detection of porcine-derived ingredients from adulterated meat based on real-time loop-mediated isothermal amplification.

Increasingly globalized and complex food supply chains contribute to a growing problem of meat fraud. Meat adulteration with pork is especially exceptionable to the global population for health concern and religious faith reasons. To prevent unfair competition and protect consumer rights, an efficient and rapid assay to identify the species of meat products is crucial. In this study, a real-time loop-mediated isothermal amplification (real-time LAMP) assay was developed for the detection of a porcine gene in meat products. The designed primers were highly selective for the porcine gene. The amplification showed no cross-reactivity with 11 other meats. The established method required 20 min with an initial amplification curve of approximately 10 min and demonstrated a detection limit of 1.76 pg/µL porcine DNA, which is 1000 times more sensitive than PCR. This study is the first attempt at detecting porcine-derived ingredients using a real-time LAMP assay in commercial products. This method meets specificity, rapidness, robustness, and sensitivity criteria; its practical application will greatly aid in battling adulteration in the food industry.

Whole-lesion histogram analysis of multiple diffusion metrics for differentiating lung cancer from inflammatory lesions.

Background: Whole-lesion histogram analysis can provide comprehensive assessment of tissues by calculating additional quantitative metrics such as skewness and kurtosis; however, few studies have evaluated its value in the differential diagnosis of lung lesions. Purpose: To compare the diagnostic performance of conventional diffusion-weighted imaging (DWI), intravoxel incoherent motion (IVIM) magnetic resonance imaging (MRI) and diffusion kurtosis imaging (DKI) in differentiating lung cancer from focal inflammatory lesions, based on whole-lesion volume histogram analysis. Methods: Fifty-nine patients with solitary pulmonary lesions underwent multiple b-values DWIs, which were then postprocessed using mono-exponential, bi-exponential and DKI models. Histogram parameters of the apparent diffusion coefficient (ADC), true diffusivity (D), pseudo-diffusion coefficient (D*), and perfusion fraction (f), apparent diffusional kurtosis (Kapp) and kurtosis-corrected diffusion coefficient (Dapp) were calculated and compared between the lung cancer and inflammatory lesion groups. Receiver operating characteristic (ROC) curves were constructed to evaluate the diagnostic performance. Results: The ADCmean, ADCmedian, D mean and D median values of lung cancer were significantly lower than those of inflammatory lesions, while the ADCskewness, Kapp mean, Kapp median, Kapp SD, Kapp kurtosis and Dapp skewness values of lung cancer were significantly higher than those of inflammatory lesions (all p < 0.05). ADCskewness (p = 0.019) and D median (p = 0.031) were identified as independent predictors of lung cancer. D median showed the best performance for differentiating lung cancer from inflammatory lesions, with an area under the ROC curve of 0.777. Using a D median of 1.091 × 10-3 mm2/s as the optimal cut-off value, the sensitivity, specificity, positive predictive value and negative predictive value were 69.23%, 85.00%, 90.00% and 58.62%, respectively. Conclusions: Whole-lesion histogram analysis of DWI, IVIM and DKI parameters is a promising approach for differentiating lung cancer from inflammatory lesions, and D median shows the best performance in the differential diagnosis of solitary pulmonary lesions.

Rhodamine-based highly sensitive colorimetric off-on fluorescent chemosensor for Hg2+ in aqueous solution and for live cell imaging.

A novel rhodamine-based highly sensitive and selective colorimetric off-on fluorescent chemosensor for Hg(2+) ions is designed and prepared by using the well-known thiospirolactam rhodamine chromophore and furfural hydrazone as signal-reporting groups. The photophysical characterization and Hg(2+)-binding properties of sensor RS1 in neutral N, N-dimethylformamide (DMF) aqueous solution are also investigated. The signal change of the chemosensor is based on a specific metal ion induced reversible ring-opening mechanism of the rhodamine spirolactam. The response of the chemosensor for Hg(2+) ions is instantaneous and reversible. And it successfully exhibits a remarkably "turn on" response toward Hg(2+) over other metal ions (even those that exist in high concentration). Moreover, this sensor is applied for in vivo imaging in Rat Schwann cells to confirm that RS1 can be used as a fluorescent probe for monitoring Hg(2+) in living cells with satisfying results, which further demonstrates its value of practical applications in environmental and biological systems.

Carbonyl-rich Poly(pyrene-4,5,9,10-tetraone Sulfide) as Anode Materials for High-Performance Li and Na-Ion Batteries.

Organic carbonyl electrode materials are widely employed for alkali metal-ion secondary batteries in terms of their sustainability, structure designability and abundant resources. As a typical redox-active organic electrode materials, pyrene-4, 5, 9, 10-tetraone (PT) shows high theoretical capacity due to the rich carbonyl active sites. But its electrochemical behavior in secondary batteries still needs further exploration. Herein, PT-based linear polymers (PPTS) is synthesized with thioether bond as bridging group and then employed as an anode material for lithium-ion batteries (LIBs) and sodium-ion batteries (SIBs). As expected, PPTS shows improved conductivity and insolubility in the non-aqueous electrolyte. When used as an anode material for LIBs, PPTS delivers a high reversible specific capacity of 697.1 mAh g-1 at 0.1 A g-1 and good rate performance (335.4 mAh g-1 at 1 A g-1 ). Moreover, a reversible specific capacity of 205.2 mAh g-1 at 0.05 A g-1 could be obtained as an anode material for SIBs.

Real-time loop-mediated isothermal amplification for rapid detection of Enterocytozoon hepatopenaei.

BACKGROUND: Enterocytozoon hepatopenaei (EHP) is a newly emerged microsporidian parasite that causes retarded shrimp growth in many countries. But there are no effective approaches to control this disease to date. The EHP could be an immune risk factor for increased dissemination of other diseases. Further, EHP infection involves the absence of obvious clinical signs and it is difficult to identify the pathogen through visual examination, increasing the risk of disease dissemination. It is urgent and necessary to develop a specific, rapid and sensitive EHP-infected shrimp diagnostic method to detect this parasite. In the present study, we developed and evaluated a rapid real-time loop-mediated isothermal amplification (real-time LAMP) for detection of EHP. METHODS: A rapid and efficient real-time LAMP method for the detection of EHP has been developed. Newly emerged EHP pathogens in China were collected and used as the sample, and three sets of specificity and sensitivity primers were designed. Three other aquatic pathogens were used as templates to test the specificity of the real-time LAMP assay. Also, we compared the real-time LAMP with the conventional LAMP by the serial dilutions of EHP DNA and their amplification curves. Application of real-time LAMP was carried out with clinical samples. RESULTS: Positive products were amplified only from EHP, but not from other tested species, EHP was detected from the clinical samples, suggesting a high specificity of this method. The final results of this assay were available within less than 45 min, and the initial amplification curve was observed at about 6 min. We found that the amplification with an exponential of sixfold dilutions of EHP DNA demonstrated a specific positive signal by the real-time LAMP, but not for the LAMP amplicons from the visual inspection. The real-time LAMP amplification curves demonstrated a higher slope than the conventional LAMP. DISCUSSION: In this study, pathogen virulence impacts have been increased in aquaculture and continuous observation was predominantly focused on EHP. The present study confirmed that the real-time LAMP assay is a promising and convenient method for the rapid identification of EHP in less time and cost. Its application greatly aids in the detection, surveillance, and prevention of EHP.

Quantitative analysis on collagen morphology in aging skin based on multiphoton microscopy.

Multiphoton microscopy was employed for monitoring the structure changes of mouse dermis collagen in the intrinsic- or the extrinsic-age-related processes in vivo. The characteristics of textures in different aging skins were uncovered by fast Fourier transform in which the orientation index and bundle packing of collagen were quantitatively analyzed. Some significant differences in collagen-related changes are found in different aging skins, which can be good indicators for the statuses of aging skins. The results are valuable to the study of aging skin and also of interest to biomedical photonics.

Preparation of quantum dots encoded microspheres by electrospray for the detection of biomolecules.

In this paper, a novel method based on the electrospray technique has been developed for preparation of quantum dot (QD)-encoded microspheres for the fist time. By electrospraying the mixture of polymer solution and quantum dots solution (single-color QDs or multi-color QDs), it is accessible to obtain a series of composite microspheres containing the functional nanoparticle. Poly(styrene-acrylate) was utilized as the electrospray polymer materials in order to obtain the microsphere modified with carboxyl group on the surface. Moreover, to test the performance of the QD-encoded microsphere in bioapplication, it is carried out that immunofluorescence analysis between antigens of mouse IgG immobilized on the functional microsphere and FITC labeled antibodies of goat-anti-mouse IgG in experiment. To the best of our knowledge, this is the first report of QD-encoded microspheres prepared by electrospray technology. This technology can carry out the one-pot preparation of different color QD-encoded microspheres with multiple intensities. This technology could be also suitable for encapsulating other optical nanocrystals and magnetic nanoparticles for obtaining multifunctional microspheres. All of the results in this paper show that the fluorescence beads made by electrospray technique can be well applied in multiplex analysis. These works provide a good foundation to accelerate application of preparing microspheres by electrospray technique in practice.

Electrospun novel bifunctional magnetic-photoluminescent nanofibers based on Fe2O3 nanoparticles and europium complex.

Novel bifunctional magnetic-photoluminescent nanofibers based on Fe(2)O(3) nanoparticles and europium complex Eu(DBM)(3)(Bath) (DBM=dibenzoylmethanate, Bath=bathophenanthroline) have been prepared by electrospinning. Extensive characterizations of the resulting bifunctional nanofibers have been performed using X-ray diffraction (XRD), field-emission scanning electron microscopy (FESEM), energy dispersive X-ray (EDX) analysis, transmission electron microscopy (TEM), and vibrating sample magnetometer (VSM). The influence on photoluminescence properties of bifunctional nanofibers of the addition of Fe(2)O(3) nanoparticles has also been studied. The results indicate that due to decreased symmetry in the composite nanofibers the excitation bands of the composite nanofibers are split into two different components. Furthermore, the existence of the Fe(2)O(3) and polymer hybrid matrixes can improve the thermal and photo stability of the europium complex and elongate the fluorescence lifetime of the europium complex.

Synthesis of magnetic and fluorescent multifunctional hollow silica nanocomposites for live cell imaging.

In this paper, we report a synthesis of multifunctional core/shell silica nanocomposites in mixed water-ethanol solvents at room temperature. Water-soluble CTAB-stabilized nanoparticles (Fe(3)O(4) and quantum dots) are used as templates and tetraethoxysilane (TEOS) is used as a precursor to fabricate multifunctional hollow silica nanocomposites. Owing to the high abundance of folate receptors in many cancer cells, folic acid is used as the targeting ligand. By coupling with folic acids, the multifunctional silica nanocomposites conjugates are successfully used for tumor cell imaging. In vitro cellular uptakes of such SiO(2) nanocomposites are investigated with fluorescence microscope, which demonstrate much higher internalization of the folate-decorated SiO(2) nanocomposites by Hela cancer cells which are of over-expression of folate receptors than the cellular uptake by NIH 3T3 fibroblast cells which are of low expression of folate receptors. Magnetic manipulation, fluorescence imaging, hollow structure, and cell targeting are simultaneously possible using a multifunctional silica nanocomposite. Our results demonstrate a robust hydrophobic nanoparticles-based approach for preparing multifunctional and biocompatible hollow silica composites, which could be also suitable for silica coating of other kinds of nanoparticles.

Aggregation states of rhodamine 6G in electrospun nanofibrous films.

Novel fluorescent composite nanofibrous films of rhodamine 6G (Rh6G) and polyacrylonitrile (PAN) are first prepared by electrospinning. The aggregation states of Rh6G in electruspun nanofibrous films are studied as a function of concentrations and characterized by UV-vis absorption spectroscopy and emission and excitation fluorescence spectroscopy. We have also used casting films as reference material to compare the effect of incorporation of Rh6G in electrospun nanofibrous films and casting films. The large specific surface area of the nanofibers and fast evaporation of the solvents in the electrospinning process reduced the aggregation of Rh6G. The appearance of fluorescent J-type dimers, even at higher dye concentration in elctrospun films, demonstrates that the electrospun films are an ideal material for incorporation of fluorescent dyes.