Aptamers for nanobodies: A nontoxic alternative to toxic ochratoxin A in immunoassays.

To measure toxins using immunoassays, hazardous toxin standards need to be added for quantification. To solve this problem, we propose to use aptamers as competitors to replace toxin standards. In this work, aptamers specific for ochratoxin A (OTA) nanobodies were selected using a DNA library containing a 36 nucleotide random region. The obtained sequences were highly aligned and the best competitor was identified to be a sequence named apt2-OT based on an indirect competitive enzyme-linked immunosorbent assay (ELISA). The Kd of apt2-OT was measured to be 2.86 µM using local surface plasmon resonance spectroscopy. The optimal apt2-OT was identified to substitute the OTA standard with a concentration needed for 50% inhibition of binding (IC50) of 3.26 µM based on a nontoxic direct competitive ELISA. The equivalence relationship between the aptamer and OTA was established in a flour sample, and a recovery experiment was performed. The detection limit for this method was 0.23 ng/mL, with a linear range from 0.25 to 10.50 ng/mL. The recovery rate was 97.5%-115.5%. This study provides a low-cost, rapid and environmentally friendly alternative to the development of immunoassays for toxins.

Zebrafish as model organisms for toxicological evaluations in the field of food science.

Food safety has long been an area of concern. The selection of stable and efficient model organisms is particularly important for food toxicology studies. Zebrafish (Danio rerio) are small model vertebrates, and 70% of human genes have at least one zebrafish ortholog. Zebrafish have advantages as model organisms due to their short life cycle, strong reproductive ability, easy rearing, and low cost. Zebrafish embryos have the advantage of being sensitive to the breeding environment and thus have been used as biosensors. Zebrafish and their embryos have been widely used for food toxicology assessments. This review provides a systematic and comprehensive summary of food toxicology studies using zebrafish as model organisms. First, we briefly introduce the multidimensional mechanisms and structure-activity relationship studies of food toxicological assessment. Second, we categorize these studies according to eight types of hazards in foods, including mycotoxins, pesticides, antibiotics, heavy metals, endocrine disruptors, food additives, nanoparticles, and other food-related ingredients. Finally, we list the applications of zebrafish in food toxicology studies in line with future research prospects, aiming to provide a valuable reference for researchers in the field of food science.

Selection of a Novel DNA Aptamer Specific for 5-Hydroxymethylfurfural Using Capture-SELEX.

A capture systematic evolution of ligands by exponential enrichment (Capture-SELEX) was described to discover novel aptamers specific for 5-hydroxymethylfurfural (5-HMF), and a biosensor based on molecular beacon was constructed to detect 5-HMF. The ssDNA library was immobilized to streptavidin (SA) resin to select the specific aptamer. The selection progress was monitored using real-time quantitative PCR (Q-PCR), and the enriched library was sequenced by high-throughput sequencing (HTS). Candidate and mutant aptamers were selected and identified by Isothermal Titration Calorimetry (ITC). The FAM-aptamer and BHQ1-cDNA were designed as the quenching biosensor to detect 5-HMF in milk matrix. After the 18th round selection, the Ct value decreased from 9.09 to 8.79, indicating that the library was enriched. The HTS results indicated that the total sequence numbers for 9th, 13th, 16th, and 18th were 417054, 407987, 307666, and 259867, but the number of sequences for the top 300 sequences was gradually increased from 9th to 18th, and the ClustalX2 analysis showed that there were four families with high homology rate. ITC results indicated that the Kd values of H1 and its mutants H1-8, H1-12, H1-14, and H1-21 were 2.5 µM, 1.8 µM, 1.2 µM, 6.5 µM, and 4.7 µM. The linear range of the quenching biosensor was from 0 µM to 75 µM, and it had a similar linear range in the 0.1% milk matrix. This is the first report to select a novel aptamer specific for 5-HMF and develop quenching biosensor for the rapid detection of 5-HMF in milk matrix.

Comparison of the Effects of 5-Hydroxymethylfurfural in Milk Powder Matrix and Standard Water on Oxidative Stress System of Zebrafish.

5-Hydroxymethylfurfural (5-HMF) and furfural (FF) are products of the maillard reaction (MR) in milk powder and their safety is controversial. The concentration changes of 5-HMF and FF after a period of cold storage were determined by high-performance liquid chromatography (HPLC). Then, we compared the toxicity effects of 5-HMF (2, 20, or 200 µM) in milk powder matrix and standard water on the oxidative stress system of zebrafish embryos. The results showed that the concentration of 5-HMF was stable, and the concentration of FF degraded over time. 5-HMF-exposed zebrafish embryos had a LC50 value of 961 µM for 120 h. High-concentration of 5-HMF exposure resulted in developmental toxicity and induced oxidative stress. 5-HMF exposure resulted in low expression of gstr gene at 200 µM in both matrices. Moreover, sod, cat, gstr, and gpxla genes were differentially highly expressed in other groups or showed no significant difference. Residual levels in all groups were well below the exposed dose, with a maximum value of only 0.4‱. These results provided a theoretical basis for understanding the effects of 5-HMF exposure in milk powder matrix on the oxidative stress system and suggested that the presence of 5-HMF in our daily consumption of milk powder does not produce significant toxic effects and need not be overstressed.

[Climatic suitability model for spring maize in Northeast China].

Crop growth is a continuous and dynamic process, and thus, the optimal meteorological indices for crop growth also change continuously. In this paper, the estimation algorithms of the daily available precipitation and the temporal interpolation methods of meteorological indices for crop growth were developed, and the climatic suitability model at daily scale was established, which was tested by the spring maize data of agro-meteorological stations in Northeast China. The simulated climatic suitability had significantly positive correlations with the height and 100-grain mass of spring maize. The correlation coefficient (R2) between the climatic suitability and the spring maize height was over 0.58 at vegetative growth stage, and over 0.45 at reproductive growth stage. The established climatic suitability model at daily scale could objectively reflect the interactions between crop growth and meteorological factors at vegetative growth stage and reproductive growth stage.

[An operational remote sensing algorithm of land surface evapotranspiration based on NOAA PAL dataset].

Based on the time series 10-day composite NOAA Pathfinder AVHRR Land (PAL) dataset (8 km x 8 km), and by using land surface energy balance equation and "VI-Ts" (vegetation index-land surface temperature) method, a new algorithm of land surface evapotranspiration (ET) was constructed. This new algorithm did not need the support from meteorological observation data, and all of its parameters and variables were directly inversed or derived from remote sensing data. A widely accepted ET model of remote sensing, i. e., SEBS model, was chosen to validate the new algorithm. The validation test showed that both the ET and its seasonal variation trend estimated by SEBS model and our new algorithm accorded well, suggesting that the ET estimated from the new algorithm was reliable, being able to reflect the actual land surface ET. The new ET algorithm of remote sensing was practical and operational, which offered a new approach to study the spatiotemporal variation of ET in continental scale and global scale based on the long-term time series satellite remote sensing images.

[Dynamic assessment of grassland degradation in Naqu of northern Tibet].

Based on the past years vegetation cover, annual maximal grass yield and June-September mean modified soil-adjusted vegetation index (MSAVI) that were inversely deduced from the 10-day composite data of NOAA/AVHRR channels 1 and 2 and NDVI in 1982-2000, this paper analyzed the recent 20 years dynamics of grassland desertification in Naqu of northern Tibet. The results showed that in recent 20 years, the area of degraded grassland in Naqu was averagely 43.1% of the total land area. It was decreased in the former ten years while increased in the latter ten years, but overall, had a decreasing trend. The degraded area was larger in west part of Naqu than in its other regions. Among the eight climatic factors including temperature, precipitation, potential evapotranspiration, vapor pressure, wind velocity, sunshine hour, ratio of precipitation to evapotranspiration, and ratio of temperature to precipitation, the most remarkable factor affecting the dynamic of grassland degradation was the potential evapotranspiration.

[Classification and dynamic changes of grasslands in northern Tibet based on recent 20 years satellite data].

Based on the NOAA/AVHRR 10-day composite NDVI data from 1982 to 2000 and by the methods of principal component analysis and unsupervised classification, the vegetations in Naqu District of northern Tibet were classified, and the annual and inter-annual variations of NDVI on the pixels selected from different grassland types were analyzed. The regions where the mean NDVI values during the main growth season of grass were equal to or great than 0.1 were defined as vegetation regions, while those where the mean NDVI values were less than 0.1 were defined as non-vegetation regions. The spatiotemporal changes of NDVI in each pixel in vegetation regions were analyzed, and the results showed that the grasslands in northern Tibet could be classified into four types, i. e., high-cold meadow, high-cold meadow steppe, high-cold steppe, and high-cold desert. This classification was accorded with practical status. The annual changes of the NDVI in the four grassland types showed one peak value, with the maximum in August. In recent 20 years, the mean NDVI from July to August in vegetation regions decreased gradually from southeast to northwest, ranging from 0.1 to 0.6, and the variation coefficient changed between 0.05 and 0.40, which was smaller in the place of high NDVI than in the place of low NDVI. The annual variability of NDVI changed between -0.005 and 0.008. In recent 20 years, the vegetations had no observable change, but the vegetation development in 20% of vegetation regions, which mainly located in Nima in western Naqu and in Jiali, Biru, Suoxian and Baqing in eastern Naqu, was weakened.

[Variation trends of China terrestrial vegetation net primary productivity and its responses to climate factors in 1982-2000].

A new estimation model of vegetation net primary production (NPP) based on remote sensing data and climatic data was presented, with which, the NPP of China terrestrial vegetation in 1982-2000 was estimated, and the intra- and inter- annual variation patterns of the NPP and its responses to climate factors were studied. The results showed that there was an obvious seasonal regularity in the intra-annual variation of the NPP. In 1982-2000, all the terrestrial vegetation types presented an increasing annual NPP, with the greatest increment for deciduous needle leaf forests and the smallest one for grasses. Evergreen broadleaf forests had the largest inter-annual variation, while grasses had the smallest one. Comparing with temperature, precipitation played a stronger driving role in the intra-annual variation of the NPP, and the effects of precipitation and temperature were more obvious in North China than in South China. The driving roles of the climate factors varied with season and latitude.