Heterologous Expression and Characterization of a pH-Stable Chitinase from Micromonospora aurantiaca with a Potential Application in Chitin Degradation.

To promote the bioconversion of marine chitin waste into value-added products, we expressed a novel pH-stable Micromonospora aurantiaca-derived chitinase, MaChi1, in Escherichia coli and subsequently purified, characterized, and evaluated it for its chitin-converting capacity. Our results indicated that MaChi1 is of the glycoside hydrolase (GH) family 18 with a molecular weight of approximately 57 kDa, consisting of a GH18 catalytic domain and a cellulose-binding domain. We recorded its optimal activity at pH 5.0 and 55 °C. It exhibited excellent stability in a wide pH range of 3.0-10.0. Mg2+ (5 mM), and dithiothreitol (10 mM) significantly promoted MaChi1 activity. MaChi1 exhibited broad substrate specificity and hydrolyzed chitin, chitosan, cellulose, soluble starch, and N-acetyl chitooligosaccharides with polymerization degrees ranging from three to six. Moreover, MaChi1 exhibited an endo-type cleavage pattern, and it could efficiently convert colloidal chitin into N-acetyl-D-glucosamine (GlcNAc) and (GlcNAc)2 with yields of 227.2 and 505.9 mg/g chitin, respectively. Its high chitin-degrading capacity and exceptional pH tolerance makes it a promising tool with potential applications in chitin waste treatment and bioactive oligosaccharide production.

Biochemical and molecular-level effects of co-exposure to chlorpyrifos and lambda-cyhalothrin on the earthworm (Eisenia fetida).

Farmland soil organisms frequently encounter pesticide mixtures presented in their living environment. However, the underlying toxic mechanisms employed by soil animals to cope with such combined pollution have yet to be explored. This investigation aimed to reveal the changes in cellular and mRNA levels under chlorpyrifos (CPF) and lambda-cyhalothrin (LCT) co-exposures in earthworms (Eisenia fetida). Results exhibited that the combination of CPF and LCT triggered an acute synergistic influence on the animals. Most exposures resulted in significant alterations in the activities of total superoxide dismutase (T-SOD), copper/zinc superoxide dismutase (Cu/Zn-SOD), caspase 3, and carboxylesterase (CarE) compared to the basal level. Moreover, when exposed to chemical mixtures, the transcription levels of four genes [heat shock protein 70 (hsp70), gst, sod, and calreticulin (crt)] also displayed more pronounced changes compared with their individual exposures. These changes in determined parameters indicated the occurrence of oxidative stress, cell death, detoxification dysfunction, and endoplasmic reticulum damage after co-exposure to CPF and LCT in E. fetida. The comprehensive examination of mixture toxicities of CPF and LCT at different endpoints would help to understand the overall toxicity they cause to soil invertebrates. The augmented deleterious effect of these pesticides in a mixture suggested that mixture toxicity assessment was necessary for the safety evaluation and application of pesticide mixtures.

Dual Functional Ultrasensitive Point-of-Care Clinical Diagnosis Using Metal-Organic Frameworks-Based Immunobeads.

Sepsis is an acute systemic infectious syndrome with high fatality. Fast and accurate diagnosis, monitoring, and medication of sepsis are essential. We exploited the fluorescent metal-AIEgen frameworks (MAFs) and demonstrated the dual functions of protein detection and bacteria identification: (i) ultrasensitive point-of-care (POC) detection of sepsis biomarkers (100 times enhanced sensitivity); (ii) rapid POC identification of Gram-negative/positive bacteria (selective aggregation within 20 min). Fluorescent lateral flow immunoassays (LFAs) are convenient and inexpensive for POC tests. MAFs possess a large surface area, excellent photostability, high quantum yield (∼80%), and multiple active sites serving as protein binding domains for ultrasensitive detection of sepsis biomarkers (IL-6/PCT) on LFAs. The limit of detection (LOD) for IL-6/PCT is 0.252/0.333 pg/mL. Rapid appraisal of infectious bacteria is vital to guide the use of medicines. The dual-functional fluorescent MAFs have great potential in POC tests for the clinical diagnosis of bacterial infections.

Automated Microfluidic Nucleic Acid Detection Platform-Integrated RPA-T7-Cas13a for Pathogen Diagnosis.

There is a growing urgent need for point-of-care testing (POCT) devices that integrate sample pretreatment and nucleic acid detection in a rapid, economical, and non-labor-intensive way. Here, we have developed an automated, portable nucleic acid detection system employing microfluidic chips integrating rotary valve-assisted sample pretreatment and recombinase polymerase amplification (RPA)-T7-Cas13a into one-step nucleic acid detection. The RPA and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas13a were integrated into a single-chamber reaction. As a validation model, we used this method to detect Group B streptococci (GBS) DNA and achieved a detection sensitivity of 8 copies/reaction, which is 6 times more sensitive than gold-standard polymerase chain reactions (PCRs). Dual specific recognition of RPA with CRISPR/Cas13a makes our method ultraspecific, with correct detection of Group B streptococci from 8 kinds of pathogenic bacteria. For the 16 positive and 24 negative clinical GBS samples, our assay achieved 100% accuracy compared to the PCR technique. The whole procedure can be automatically completed within 30 min, providing a more robust, sensitive, and accurate molecular diagnostic tool for POCT.

Recent advancements in nucleic acid detection with microfluidic chip for molecular diagnostics.

The coronavirus disease 2019 (COVID-19) has extensively promoted the application of nucleic acid testing technology in the field of clinical testing. The most widely used polymerase chain reaction (PCR)-based nucleic acid testing technology has problems such as complex operation, high requirements of personnel and laboratories, and contamination. The highly miniaturized microfluidic chip provides an essential tool for integrating the complex nucleic acid detection process. Various microfluidic chips have been developed for the rapid detection of nucleic acid, such as amplification-free microfluidics in combination with clustered regularly interspaced short palindromic repeats (CRISPR). In this review, we first summarized the routine process of nucleic acid testing, including sample processing and nucleic acid detection. Then the typical microfluidic chip technologies and new research advances are summarized. We also discuss the main problems of nucleic acid detection and the future developing trend of the microfluidic chip.

Dual Gold Nanoparticle/Chemiluminescent Immunoassay for Sensitive Detection of Multiple Analytes.

Multiple antibiotics and mycotoxins usually simultaneously exist in foods, which poses a serious threat to human health. How to detect them in one test with high sensitivity and fidelity is challenging. In this study, we develop a dual readout lateral flow immunodetection platform that can quantitatively detect five kinds of antibiotics and five kinds of mycotoxins within one sample. The platform is composed of a chip and a portable readout instrument where gold nanoparticle (AuNP)-based and chemiluminescence immunoassays could be performed to reach a maximum throughput of 220 analytes in one setting. For a rapid screen, qualitative analysis by detecting the color change of the deposited AuNPs on the chip could be realized. For quantitative results, chemiluminescence imaging and analysis can be completed within 15 min. Apart from the high throughput and high efficiency, this platform has a high detection sensitivity. For instance, the limit of detection (LOD) for thiamphenicol (a representative antibiotic) and fumonisins B1 (a representative mycotoxin) is 8 times and 40 times lower than those of the previously reported methods, respectively. Thus, this dual readout immunodetection platform is promising as a universal device for rapid and quantitative detection of multiple analytes with high throughput, high sensitivity, and high fidelity.

PRAPNet: A Parallel Residual Atrous Pyramid Network for Polyp Segmentation.

In a colonoscopy, accurate computer-aided polyp detection and segmentation can help endoscopists to remove abnormal tissue. This reduces the chance of polyps developing into cancer, which is of great importance. In this paper, we propose a neural network (parallel residual atrous pyramid network or PRAPNet) based on a parallel residual atrous pyramid module for the segmentation of intestinal polyp detection. We made full use of the global contextual information of the different regions by the proposed parallel residual atrous pyramid module. The experimental results showed that our proposed global prior module could effectively achieve better segmentation results in the intestinal polyp segmentation task compared with the previously published results. The mean intersection over union and dice coefficient of the model in the Kvasir-SEG dataset were 90.4% and 94.2%, respectively. The experimental results outperformed the scores achieved by the seven classical segmentation network models (U-Net, U-Net++, ResUNet++, praNet, CaraNet, SFFormer-L, TransFuse-L).

EIEN: Endoscopic Image Enhancement Network Based on Retinex Theory.

In recent years, deep convolutional neural network (CNN)-based image enhancement has shown outstanding performance. However, due to the problems of uneven illumination and low contrast existing in endoscopic images, the implementation of medical endoscopic image enhancement using CNN is still an exploratory and challenging task. An endoscopic image enhancement network (EIEN) based on the Retinex theory is proposed in this paper to solve these problems. The structure consists of three parts: decomposition network, illumination correction network, and reflection component enhancement algorithm. First, the decomposition network model of pre-trained Retinex-Net is retrained on the endoscopic image dataset, and then the images are decomposed into illumination and reflection components by this decomposition network. Second, the illumination components are corrected by the proposed self-attention guided multi-scale pyramid structure. The pyramid structure is used to capture the multi-scale information of the image. The self-attention mechanism is based on the imaging nature of the endoscopic image, and the inverse image of the illumination component is fused with the features of the green and blue channels of the image to be enhanced to generate a weight map that reassigns weights to the spatial dimension of the feature map, to avoid the loss of details in the process of multi-scale feature fusion and image reconstruction by the network. The reflection component enhancement is achieved by sub-channel stretching and weighted fusion, which is used to enhance the vascular information and image contrast. Finally, the enhanced illumination and reflection components are multiplied to obtain the reconstructed image. We compare the results of the proposed method with six other methods on a test set. The experimental results show that EIEN enhances the brightness and contrast of endoscopic images and highlights vascular and tissue information. At the same time, the method in this paper obtained the best results in terms of visual perception and objective evaluation.

Modeling of Flame Retardants in Typical Urban Indoor Environments in China during 2010-2030: Influence of Policy and Decoration and Implications for Human Exposure.

Novel flame retardants (FRs) are of increasing concern, given growing evidence of health effects and use to replace polybrominated diphenyl ethers (PBDEs). This study modeled combined effects of use policies and decoration on indoor FRs and human exposure for 18 widely used PBDEs, organophosphate esters (OPEs), and novel brominated flame retardants in typical urban indoor environments in China. The current estimated indoor emission rates and average concentrations in air and dust of the 18 FRs were 102-103 ng/h, 561 ng/m3, and 1.5 × 104 ng/g, respectively, with seven OPEs dominant (>69%). Different use patterns exist between China and the US and Europe. Scenarios modeled over 2010-2030 suggested that decoration would affect indoor concentrations of FRs more than use policies, and use policies were mainly responsible for shifts of FR composition. Additional use of hexabromobenzene and 2,3,4,5,6-pentabromotoluene and removal of BDE-209 would make the total human exposure to the modeled FR mixture increase after the restriction of penta- and octa-BDE but decrease after deca-BDE was banned. Better knowledge of the toxicity of substitutes is needed for a complete understanding of the health implications of such changes. Toddlers may be more affected by use changes than adults. Such studies are supportive to the management of FR use.

Novel concept of the smart NIR-light-controlled drug release of black phosphorus nanostructure for cancer therapy.

A biodegradable drug delivery system (DDS) is one the most promising therapeutic strategies for cancer therapy. Here, we propose a unique concept of light activation of black phosphorus (BP) at hydrogel nanostructures for cancer therapy. A photosensitizer converts light into heat that softens and melts drug-loaded hydrogel-based nanostructures. Drug release rates can be accurately controlled by light intensity, exposure duration, BP concentration, and hydrogel composition. Owing to sufficiently deep penetration of near-infrared (NIR) light through tissues, our BP-based system shows high therapeutic efficacy for treatment of s.c. cancers. Importantly, our drug delivery system is completely harmless and degradable in vivo. Together, our work proposes a unique concept for precision cancer therapy by external light excitation to release cancer drugs. If these findings are successfully translated into the clinic, millions of patients with cancer will benefit from our work.

Variations of enzymatic activity and gene expression in zebrafish (Danio rerio) embryos co-exposed to zearalenone and fumonisin B1.

The natural co-occurrence of multiple mycotoxins has been reported in cereals and cereal products worldwide. Even though the dietary exposure to mycotoxins constitutes a serious human health, most reports are limited to the toxic effect of individual mycotoxins. The purpose of the present study was to assess the combined toxic effects of zearalenone (ZEN) and fumonisin B1 (FB1) and the potential interaction of their mixture on zebrafish (Danio rerio) embryos. Our results showed that ZEN possessed the higher toxicity to embryonic zebrafish (7-day LC50 value of 0.78 mg a.i. L-1) compared with FB1 (7-day LC50 value of 227.7 mg a.i. L-1). The combination of ZEN and FB1 exerted an additive effect on zebrafish embryos. Meanwhile, the activities of antioxidant CAT, caspase-3, and detoxification enzyme CYP450, as well as the expressions of six genes (Mn-sod, cas9, bax, cc-chem, ERα, and crh) associated with oxidative stress, cellular apoptosis, immune system, and endocrine system were prominently altered in the mixture exposure compared with the corresponding single treatment group of ZEN or FB1. Taken together, the regulatory standards of mycotoxins in food and feed should be updated based on the mixture effects of mycotoxins, and there is an increased need on effective detoxification methods for controlling and reducing the toxicity of multiple mycotoxins in animal feed and throughout the food supply chain.

Occurrence and human exposure assessment of organophosphate esters in atmospheric PM2.5 in the Beijing-Tianjin-Hebei region, China.

Organophosphate esters (OPEs) in atmospheric fine particles (PM2.5) were comprehensively investigated in the Beijing-Tianjin-Hebei (BTH) region from April 2016 to March 2017. The concentrations of Σ8OPEs in all the five sampling sites ranged from 90 to 8291 pg/m3 (mean 1148 ± 1239 pg/m3; median 756 pg/m3). The highest level (median 1067 pg/m3) was found at one of the urban sites in Beijing, followed by Tianjin (746 pg/m3) and Shijiazhuang (724 pg/m3). Tris(2-chloroethyl) phosphate (TCEP) and tri[(2R)-1-chloro-2-propyl] phosphate (TCPP) were the dominant compounds across the five sampling locations. Generally, the concentrations of chlorinated OPEs were relatively higher in summer than in winter (p < 0.05), but no significant seasonal difference was discovered for non-chlorinated individual OPEs. The concentrations of tri-n-butyl phosphate (TBP), TCEP, TCPP and triphenyl phosphate (TPP) were positively correlated with the meteorological parameters (i.e. temperature and relative humidity) (p < 0.05), indicating an evident influence of meteorological condition on OPE distribution. We observed a negative correlation (p < 0.05) between octanol-air partition coefficients (logKoa) and the ratio of PM2.5-bound OPE concentrations to total suspended particulates-bound OPE concentrations, suggesting that physicochemical properties affect the particle-size distribution of OPEs. Furthermore, the median value of cancer hazard quotients (HQs) of TCEP was higher than TBP and tris(2-ethylhexyl) phosphate (TEHP). The health risk assessment showed that HQ values for children were ~1.6 times higher than those for adults. Relatively higher health risk induced by PM2.5-bound OPEs via inhalation was found during severe hazy days than in clear days.

Toxicological interactions of cadmium and four pesticides on early life stage of rare minnow (Gobiocypris rarus).

Although chemicals have been traditionally regulated on an individual basis in aquatic ecosystems, they often co-exist as different types of complex mixtures. Laboratory assays were conducted for assessing the responses of rare minnow (Gobiocypris rarus) to individual and mixture chemicals [trace element cadmium (Cd), thiamethoxam, deltamethrin, malathion and prochloraz]. Data obtained from 96 h semi-static toxicity assays implied that deltamethrin elicited the highest toxic effect on the various developmental phases (larval, juvenile and adult phases) of G. rarus with LC50 values ranging from 0.00061 to 0.25 mg a.i. L-1, followed by prochloraz, malathion and Cd with 96-h LC50 values ranging from 0.49 to 1.1, from 7.1 to 26, and from 7.6 to 15 mg a.i. L-1, respectively. Thiamethoxam elicited the lowest toxic effect on the organisms with 96-h LC50 values ranging from 38 to 202 mg a.i. L-1. Larval phase was not always the most sensitive period in the three detected phases to most of chemicals. Chemical combinations containing deltamethrin and malathion displayed synergetic responses to the larvae of G. rarus. Besides, the binary mixtures of Cd-deltamethrin and Cd-prochloraz also exhibited synergetic response to rare minnows. Our results indicate that extra information is necessary to develop practical criteria for selecting chemical combinations that require legislative attention according to their likelihood to exert synergetic responses. Thence, more investigations on mixture toxicities of various chemicals should be taken as a priority for producing synergetic interaction to improve the environmental risk assessment of chemicals.

Seasonal variation and human exposure assessment of legacy and novel brominated flame retardants in PM2.5 in different microenvironments in Beijing, China.

Indoor exposure to legacy and novel brominated flame retardants (NBFRs) may cause potential risks to human health. Studies on seasonal variations of indoor PM2.5-bound BFRs are scant. This study comprehensively investigated the seasonal variations of PM2.5-bound polybrominated diphenyl ethers (PBDEs) and NBFRs in various indoor environments (i.e. activity room, dormitory, home and office) and outdoor PM2.5 in Beijing, China over one year. The levels of PBDE (226 ±â€¯108 pg m-3) were higher than that of NBFRs (27.0 ±â€¯16.0 pg m-3) in all indoor environments. Decabromodiphenyl ether (BDE-209) and decabromodiphenyl ethane (DBDPE) were the most abundant BFRs. Office showed the highest mean concentrations of Σ15PBDEs (251 ±â€¯125 pg m-3) and Σ9NBFRs (33.0 ±â€¯18.0 pg m-3), which may be related to the higher number density of indoor materials. The concentrations of Σ9NBFRs and Σ15PBDE in indoor PM2.5 were found to be significantly higher than those in the corresponding outdoor PM2.5 (p < 0.05). Two to twenty-fold seasonal variations were observed for levels of PM2.5-bound BFRs during one year, and indoor concentrations increased slightly during the central-heating period (November 2016-March 2017). Seasonal variations of BFRs could be affected by temperature, relative humidity and concentrations of particle matters. The PM2.5-bound BFRs concentrations in PM2.5 were negatively correlated with temperature and relative humidity, while positively correlated with PM2.5 concentrations (p < 0.05). Atmospheric haze pollution could possibly contribute to higher levels of indoor PM2.5-bound BFRs. Human daily intake of BFRs via PM2.5 inhalation showed seasonal differences, and the highest exposure risk occurred in winter. Toddlers were assessed to be more vulnerable to indoor PM2.5-bound BFRs in all seasons. This study provided the first-hand measurements of seasonal concentrations and human exposure to PM2.5-bound BFRs in different indoor scenarios in Beijing.

Brominated flame retardants in atmospheric fine particles in the Beijing-Tianjin-Hebei region, China: Spatial and temporal distribution and human exposure assessment.

Atmospheric fine particle (PM2.5) samples were collected over a whole year (April 2016 - March 2017) across five sampling locations in the Beijing-Tianjin-Hebei (BTH) region, to investigate the occurrence of novel brominated flame retardants (NBFRs) and polybrominated diphenyl ethers (PBDEs). The concentrations of ∑9NBFRs were in the range of 0.63-104 pg/m3 (15.6 ±â€¯16.8 pg/m3) in atmospheric PM2.5, while the levels of ∑9PBDEs (excluding BDE-209) ranged from 0.05 to 19.1 pg/m3 (2.9 ±â€¯3.8 pg/m3) and BDE-209 concentrations ranged from 0.88 to 138 pg/m3 (22 ±â€¯28 pg/m3). Relatively higher levels of NBFRs and PBDEs were found at urban sampling sites in Beijing City and Shijiazhuang City. Decabromodiphenylethane (DBDPE) and BDE-209 were the dominant compounds with the relative abundances of 72% in ∑9NBFRs and 90% in ∑10PBDEs, respectively. Generally, the levels of most target BFRs in summer were lower than those in other seasons. However, there were no notable seasonal differences in levels of DBDPE and BDE-209 in atmospheric PM2.5 samples across the BTH region. Significant and positive correlations were found between the concentrations of BFRs and PM2.5. Daily human exposure via inhalation revealed that children have a higher probability of suffering from the adverse effects of BFRs than that of adults. In addition, residents living near sampling locations across the BTH region may suffer high exposure risks to BDE-209 and NBFRs.

Detection of autophagy processes during the development of nonarticulated laticifers in Euphorbia kansui Liou.

MAIN CONCLUSION: Autophagy is involved in cytoplasmic degradation through directly engulfing cytosol and organelles by autophagosomes and then fusing with lysosome-like vesicles during the development of nonarticulated laticifers in Euphorbia kansui Liou. Autophagy has been reported to play an important role in a wide range of eukaryotic organisms during responses to various abiotic and biotic stresses. However, until recently, the functions of autophagy in normal plant differentiation and development were still in their infancy. Nonarticulated laticifers, a type of secretory tissue in plants, undergo the degradation of cytosol and organelles during their development. However, little evidence of autophagy in laticifer differentiation has been provided. In the present study, using anti-ATG8 antibody-Alexa Fluor 488, Lyso-Tracker Red (LTR) and monodansylcadaverine (MDC) as markers for detecting autophagosomes, as well as autophagy-related structures, we observed that the green fluorescence of ATG8a largely colocalized with the red fluorescence of LTR and purple fluorescence of MDC and the quantity of autophagosomes experienced a trend from less to more to less during laticifer development. Additionally, we described the autophagy process during the development of nonarticulated laticifers in Euphorbia kansui Liou at the ultrastructural level in detail. In addition, further immunogold TEM studies also verified the presence of autophagosomes, autolysosomes and lysosome-like structures in laticifers. Taken together, these results suggest that autophagy contributes to the development of the nonarticulated laticifers in E. kansui Liou and that autophagosomes fuse with lysosome-like structures for degradation. These results will lay an important foundation for further studies on laticifer regulation.

Guidance receptor promotes the asymmetric distribution of exocyst and recycling endosome during collective cell migration.

During collective migration, guidance receptors signal downstream to result in a polarized distribution of molecules, including cytoskeletal regulators and guidance receptors themselves, in response to an extracellular gradient of chemotactic factors. However, the underlying mechanism of asymmetry generation in the context of the migration of a group of cells is not well understood. Using border cells in the Drosophila ovary as a model system for collective migration, we found that the receptor tyrosine kinase (RTK) PDGF/VEGF receptor (PVR) is required for a polarized distribution of recycling endosome and exocyst in the leading cells of the border cell cluster. Interestingly, PVR signaled through the small GTPase Rac to positively affect the levels of Rab11-labeled recycling endosomes, probably in an F-actin-dependent manner. Conversely, the exocyst complex component Sec3 was required for the asymmetric localization of RTK activity and F-actin, similar to that previously reported for the function of Rab11. Together, these results suggested a positive-feedback loop in border cells, in which RTKs such as PVR act to induce a higher level of vesicle recycling and tethering activity in the leading cells, which in turn enables RTK activity to be distributed in a more polarized fashion at the front. We also provided evidence that E-cadherin, the major adhesion molecule for border cell migration, is a specific cargo in the Rab11-labeled recycling endosomes and that Sec3 is required for the delivery of the E-cadherin-containing vesicles to the membrane.

Perchlorate in indoor dust and human urine in China: contribution of indoor dust to total daily intake.

Perchlorate is used in fireworks and China is the largest fireworks producer and consumer in the world. Information regarding human exposure to perchlorate is scarce in China, and exposure via indoor dust ingestion (EDI indoor dust) has rarely been evaluated. In this study, perchlorate was found in indoor dust (detection rate: 100%, median: 47.4 µg/g), human urine (99%, 26.2 ng/mL), drinking water (100%, 3.99 ng/mL), and dairy milk (100%, 12.3 ng/mL) collected from cities that have fireworks manufacturing areas (Yueyang and Nanchang) and in cities that do not have fireworks manufacturing industries (Tianjin, Shijiazhuang, Yuxi and Guilin) in China. In comparison with perchlorate levels reported for other countries, perchlorate levels in urine samples from fireworks sites and nonfireworks sites in China were higher. Median indoor dust perchlorate concentrations were positively correlated (r = 0.964, p < 0.001) with outdoor dust perchlorate levels reported previously. The total daily intake (EDI total) of perchlorate, estimated based on urinary levels, ranged from 0.090 to 27.72 µg/kg body weight (bw)/day for all studied participants; the percentage of donors who had EDI total exceeding the reference dose (RfD) recommended by the United States Environmental Protection Agency (US EPA) was 79%, 48%, and 25% for toddlers (median: 1.829 µg/kg bw/day), adults (0.669 µg/kg bw/day), and children (median: 0.373 µg/kg bw/day), respectively. Toddlers (0.258 µg/kg bw/day) had the highest median EDI indoor dust, which was 2 to 5 times greater than the EDI indoor dust calculated for other age groups (the range of median values: 0.044 to 0.127 µg/kg bw/day). Contribution of indoor dust to EDItotal was 26%, 28%, and 7% for toddlers, children, and adults, respectively. Indoor dust contributed higher percentage to EDI total than that by dairy milk (0.5-5%).

Colorimetric detection of copper(II) ion using click chemistry and hemin/G-quadruplex horseradish peroxidase-mimicking DNAzyme.

G-quadruplex-forming sequence can be formed through a copper(I) ion (Cu(+))-catalyzed click chemistry between azide- and alkyne-modified short G-rich sequences in aqueous solution, eliminating immobilization and washing steps of conventional assays. The source for Cu(+) was generated from the reduction of Cu(2+) with the reductant of sodium ascorbate. In the presence of hemin and K(+), the self-assembly of hemin/G-quadruplex structure has the activity of horseradish peroxidase (HRP), which can catalyze its colorless substrate tetrazmethyl benzidine (TMB) into a colored product. Hence, the concentration of Cu(2+) can be evaluated visually for qualitative analysis according to the color change of the solution, and the optical density (OD) value of the resulting solution at 450 nm was also recorded using a microplate reader for quantitative analysis.

An array of femtoliter wells for sensitive detection of copper using click chemistry.

Sensitive detection of copper ion (Cu2+), which is of great importance for environmental pollution and human health, is crucial. In this study, we present a highly sensitive method for measuring Cu2+ in an array of femtoliter wells. In brief, magnetic beads (MBs) modified with alkyne groups were bound to the azide groups of biotin-PEG3-azide (bio-PEG-N3) via Cu+-catalyzed click chemistry. Cu+ in the click chemistry reaction was generated by reducing Cu2+ with sodium ascorbate. Following the ligation, the surface of the MBs was modified with biotin, which could be labeled with streptavidin-ß-galactosidase (SßG). The MBs complex was then suspended in ß-galactosidase substrate fluorescein-di-ß-d-galactopyranoside (FDG), and loaded into the array of femtoliter wells. The MBs sank into the wells due to gravity, and the resulting fluorescent product, generated from the reaction between SßG on the surface of the MBs and FDG, was confined within the wells. The number of fluorescent wells increased with higher Cu2+ concentrations. The bright-field and fluorescent images of the wells were acquired using an inverted fluorescent microscope. The detection limit of this assay for Cu2+ was 1 nM without signal amplification, which was 103 times lower than that of traditional fluorescence detection assays.