Surface-enhanced Raman scattering based on noble metal nanoassemblies for detecting harmful substances in food.

Residues of harmful substances in food can severely damage human health. The content of these substances in food is generally low, making detection difficult. Surface-enhanced Raman scattering (SERS), based on noble metal nanomaterials, mainly gold (Au) and silver (Ag), has exhibited excellent capabilities for trace detection of various substances. Noble metal nanoassemblies, in particular, have extraordinary flexibility and tunable optical properties, which cannot be offered by single nanoparticles (NPs). These nanoassemblies, with their various morphologies synthesized using NPs through artificially induced self-assembly or template-driven preparation, can significantly enhance the local electric field and create "hot spots" due to the gaps between adjacent NPs. Consequently, the SERS properties of NPs become more prominent, leading to improved performance in the trace detection of various substances and detection limits that are considerably lower than the current relevant standards. Noble metal nanoassemblies show promising potential in ensuring food safety. This review discusses the synthesis methods and SERS properties of noble metal nanoassemblies and then concentrates on their application in detecting biotoxins, drug residues, illegal additives, and heavy metals. The study provides valuable references for further research into the application of nanoassemblies in food safety detection.

Real-Time Compensation for SLD Light-Power Fluctuation in an Interferometric Fiber-Optic Gyroscope.

An interferometric fiber-optic gyroscope (IFOG) demodulates a rotation signal via interferometric light intensity. However, the working environments of IFOGs typically involve great uncertainty. Fluctuations in temperature, air pressure, electromagnetic field, and the power system all cause the power of the superluminescent diode (SLD) light source to fluctuate as well. In this invited paper, we studied the effects of SLD power fluctuation on the dynamic and static performance characteristics of a gyro system through the use of a light-power feedback loop. Fluctuations of 0.5 mA, 1 mA, and 5 mA in the SLD source entering the IFOG caused zero-bias stability to be 69, 135, and 679 times worse. We established an effective method to monitor power fluctuations of SLD light sources and to compensate for their effects without increasing hardware complexity or system cost. In brief, we established a real-time power-sensing and -compensating system. Experimental results showed that for every 0.1 mA increase in the fluctuation amplitude of the driving current, the zero-bias stability became 4 to 7 times worse, which could be reduced about 95% through the use of SLD power compensation.

A Novel Closed-Loop Control to Solve Light Source Power Fluctuations in the Fiber-Optic Gyroscope.

The performance of a gyroscope is directly affected by the fluctuations in the light source power (LSP) in an interferometric fiber-optic gyroscope (IFOG). Therefore, it is important to compensate for fluctuations in the LSP. When the feedback phase generated by the step wave completely cancels the Sagnac phase in real-time, the error signal of the gyroscope is linearly related to the differential signal of the LSP, otherwise, the error signal of the gyroscope is uncertain. Herein, we present two compensation methods to compensate for the error of the gyroscope when the error is uncertain, which are double period modulation (DPM) and triple period modulation (TPM). Compared with the TPM, DPM has better performance, but it increases the requirements for the circuit. TPM has lower requirements for the circuit and is more suitable for small fiber- coil applications. The experimental results show that, when the frequency of the LSP fluctuation is relatively low (1 kHz and 2 kHz), DPM and TPM do not differ significantly in terms of performance; both of them can achieve an improvement of about 95% in bias stability. When the frequency of the LSP fluctuation is relatively high (4 kHz, 8 kHz and 16 kHz), DPM and TPM can achieve about 95% and 88% improvement in bias stability, respectively.

Recent advances in simultaneous detection strategies for multi-mycotoxins in foods.

Mycotoxin contamination has become a challenge in the field of food safety testing, given the increasing emphasis on food safety in recent years. Mycotoxins are widely distributed, in heavily polluted areas. Food contamination with these toxins is difficult to prevent and control. Mycotoxins, as are small-molecule toxic metabolites produced by several species belonging to the genera Aspergillus, Fusarium, and Penicillium growing in food. They are considered teratogenic, carcinogenic, and mutagenic to humans and animals. Food systems are often simultaneously contaminated with multiple mycotoxins. Due to the additive or synergistic toxicological effects caused by the co-existence of multiple mycotoxins, their individual detection requires reliable, accurate, and high-throughput techniques. Currently available, methods for the detection of multiple mycotoxins are mainly based on chromatography, spectroscopy (colorimetry, fluorescence, and surface-enhanced Raman scattering), and electrochemistry. This review provides a comprehensive overview of advances in the multiple detection methods of mycotoxins during the recent 5 years. The principles and features of these techniques are described. The practical applications and challenges associated with assays for multiple detection methods of mycotoxins are summarized. The potential for future development and application is discussed in an effort, to provide standards of references for further research.

Clonorchis sinensis ferritin heavy chain triggers free radicals and mediates inflammation signaling in human hepatic stellate cells.

Clonorchiasis, caused by direct and continuous contact with Clonorchis sinensis, is associated with hepatobiliary damage, inflammation, periductal fibrosis, and the development of cholangiocarcinoma. Hepatic stellate cells respond to liver injury through production of proinflammatory mediators which drive fibrogenesis; however, their endogenous sources and pathophysiological roles in host cells were not determined. C. sinensis ferritin heavy chain (CsFHC) was previously confirmed as a component of excretory/secretory products and exhibited a number of extrahepatic immunomodulatory properties in various diseases. In this study, we investigated the expression pattern and biological role of CsFHC in C. sinensis. CsFHC was expressed throughout life stages of C. sinensis. More importantly, we found that treatment of human hepatic stellate cell line LX-2 with CsFHC triggered the production of free radicals via time-dependent activation of NADPH oxidase, xanthine oxidase, and inducible nitric oxide synthase. The increase in free radicals substantially promoted the degradation of cytosolic IκBα and nuclear translocation of NF-κB subunits (p65 and p50). CsFHC-induced NF-κB activation was markedly attenuated by preincubation with specific inhibitors of corresponding free radical-producing enzyme or the antioxidant. In addition, CsFHC induced an increased expression level of proinflammatory cytokines, IL-1ß and IL-6, in NF-κB-dependent manner. Our results indicate that CsFHC-triggered free radical-mediated NF-κB signaling is an important factor in the chronic inflammation caused by C. sinensis infection.

Identification, immunolocalization, and characterization analyses of an exopeptidase of papain superfamily, (cathepsin C) from Clonorchis sinensis.

Cathepsin C is an important exopeptidase of papain superfamily and plays a number of great important roles during the parasitic life cycle. The amino acid sequence of cathepsin C from Clonorchis sinensis (C. sinensis) showed 54, 53, and 49% identities to that of Schistosoma japonicum, Schistosoma mansoni, and Homo sapiens, respectively. Phylogenetic analysis utilizing the sequences of papain superfamily of C. sinensis demonstrated that cathepsin C and cathepsin Bs came from a common ancestry. Cathepsin C of C. sinensis (Cscathepsin C) was identified as an excretory/secretory product by Western blot analysis. The results of transcriptional level and translational level of Cscathepsin C at metacercaria stage were higher than that at adult worms. Immunolocalization analysis indicated that Cscathepsin C was specifically distributed in the suckers (oral sucker and ventral sucker), eggs, vitellarium, intestines, and testis of adult worms. In the metacercaria, it was mainly detected on the cyst wall and excretory bladder. Combining with the results mentioned above, it implies that Cscathepsin C may be an essential proteolytic enzyme for proteins digestion of hosts, nutrition assimilation, and immune invasion of C. sinensis. Furthermore, it may be a potential diagnostic antigen and drug target against C. sinensis infection.

Biochemical and immunological characterization of annexin B30 from Clonorchis sinensis excretory/secretory products.

Clonorchis sinensis has been classified as group I biological carcinogen for cholangiocarcinoma by the World Health Organization. Biological studies on excretory/secretory products (ESPs) enabled us to understand the pathogenesis mechanism of C. sinensis and develop new strategies for the prevention of clonorchiasis. In this study, sequence analysis showed that annexin B30 from C. sinensis (CsANXB30) is composed of four annexin repeats which were characterized by type II and III Ca(2+)-binding sites or KGD motif with the capability of Ca(2+)-binding. In addition, immunoblot assay revealed that recombinant CsANXB30 (rCsANXB30) could be recognized by the sera from rats infected with C. sinensis and the sera from rats immunized by CsESPs. Real-time PCR showed that its transcriptional level was the highest at the stage of metacercaria. Immunofluorescence assay was employed to confirm that CsANXB30 was distributed in the tegument, intestine, and egg of adult worms, as well as the tegument and vitellarium of metacercaria. rCsANXB30 was able to bind phospholipid in a Ca(2+)-dependent manner and human plasminogen in a dose-dependent manner. Moreover, cytokine and antibody measurements indicated that rats subcutaneously immunized with rCsANXB30 developed a strong IL-10 production in spleen cells and a high level of IgG1 isotype, indicating that rCsANXB30 could trigger specific humoral and cellular immune response in rats. The present results implied that CsANXB30 might be involved in a host-parasite interaction and affected the immune response of the host during C. sinensis infection.

Effect of degree of polymerization on regenerated cellulose ultrafiltration membrane performance through ZnCl2/AlCl3 aqueous solvent system.

The development of a direct method for preparing regenerated cellulose (RC) ultrafiltration membranes from cellulose is urgently needed. In this study, refined cotton was used as the raw material to successfully prepare RC ultrafiltration membranes at room temperature using a ZnCl2/AlCl3 solvent system combined with a nonsolvent-induced phase separation (NIPS) method. This solvent system effectively degrades cellulose, producing RC ultrafiltration membranes with varying degrees of polymerization (DP). The research results indicate that reducing the DP of cellulose significantly decreases the viscosity of the solution, facilitating the formation of an asymmetric, finger-like pore structures in the membrane. Furthermore, a decrease in DP slightly enlarges the surface pore size and significantly thickens the dense layer. At a DP of 250, the water flux of the DP250-ET membrane reached 630 L·m-2·h-1·bar-1, with a molecular weight cut-off (MWCO) of ~300 kDa, enabling efficient separation of viruses (LRV > 3.91) and IgG. The exposure of more hydroxy groups on the RC enhances the membrane's hydrophilicity, indicated by a water contact angle (WCA) of 39.5°. Compared to commercial polyethersulfone (PES) membranes, the DP250-ET membrane exhibited lower protein adsorption and excellent anti-fouling performance in practical applications (FRR > 80 %). Overall, this work confirms the significant potential of the eco-friendly ZnCl2/AlCl3 solvent system in the fabrication of RC ultrafiltration membranes, where the structure and performance of the membrane can be tailored by adjusting the DP of cellulose.

The development of multifunctional materials for water pollution remediation using pollen and sporopollenin.

Pollen is a promising material for water treatment owing to its renewable nature, abundant sources, and vast reserves. The natural polymer sporopollenin, found within pollen exine, possesses a distinctive layered porous structure, mechanical strength, and stable chemical properties, which can be utilized to prepare sporopollenin exine capsules (SECs). Leveraging these attributes, pollen or SECs can be used to develop water pollution remediation materials. In this review, the structure of pollen is first introduced, followed by the categorization of various methods for extracting SECs. Then, the functional expansion of pollen adsorbents, with an emphasis on their recyclability, reusability, and visual sensing capabilities, as opposed to mere functional group modification, is discussed. Furthermore, the progress made in utilizing pollen as a biological template for synthesizing catalysts is summarized. Intriguingly, pollen can also be engineered into self-propelled micromotors, enhancing its potential application in adsorption and catalysis. Finally, the challenges associated with the application of pollen in water pollution treatment are discussed. These challenges include the selection of environmentally friendly, non-toxic reagents in synthesizing pollen water remediation products and the large-scale application after synthesis. Moreover, the multifunctional synthesis and application of different water remediation products are prospected.

Identification and biochemical characterization of adenylate kinase 1 from Clonorchis sinensis.

Adenylate kinase 1 is responsible for the conversion of AMP into ADP involved in purine metabolism. In the present study, adenylate kinase 1 gene (CsADK1) was isolated from an adult cDNA library of Clonorchis sinensis, and the recombinant protein was expressed in Escherichia coli. Bioinformatics analysis implied that the putative protein contained 197 amino acids, and some residues in conservative binding sites of CsADK1 were substituted. The structure modeling analysis showed that CsADK1 was composed of a core domain, an NMP-binding domain, and a LID domain, which was just a small loop. It demonstrated that CsADK1 was a short isoform of ADKs. Moreover, CsADK1 was identified as an excretory/secretory product by western blot analysis. Real-time quantitative PCR showed that expression level of CsADK1 at the stage of excysted metacercaria was higher than those of adult worm (18.8-folds, P<0.01), metacercariae (1.5-folds, P<0.01), and eggs (5.6-folds, P<0.01). In addition, histochemistry analysis showed that CsADK1 was extensively distributed in metacercariae and in the vitellaria and eggs of adult worms. The Km and Vmax value for substrate ADP were 2.2 mM and 0.9 mM/min, respectively. The optimal temperature and pH value were 37 °C and from 7.5 to 8.0, respectively. The enzyme activity was highly dependent on Mg2+, and the optimal concentration of Mg2+ was 2 mM. However, the enzyme activity was slightly activated by Ca2+, and Mn2+ has no effect on activity. For monovalent ions, activity was highly activated by K+ and NH4+, but slightly by Li+. Taken together, CsADK1 was a metal ion-dependent enzyme involved in purine metabolism, which was important for development and reproduction, and might be a potential candidate for drug target for clonorchiasis.

Clonorchis sinensis acetoacetyl-CoA thiolase: identification and characterization of its potential role in surviving in the bile duct.

BACKGROUND: Clonorchis sinensis (C. sinensis) inhabits in bile duct of the host. However, the mechanisms involved in why C. sinensis can survive in the bile environment containing lipids have not yet been explored. In this study, C. sinensis acetoacetyl-CoA thiolase (CsACAT), a member of the thiolase family which has a key role in the beta oxidation pathway of fatty acid production, was identified and characterized to understand its potential role in adapting to the bile environment. METHODS: The encoding sequence, conserved domains and spatial structure of CsACAT were identified and analyzed by bioinformatic tools. Recombinant CsACAT (rCsACAT) was obtained using a procaryotic expression system. The expression pattern of CsACAT was confirmed by quantitative real-time PCR, western blotting, and immunofluorescence. Gradients of lecithin were then set to culture C. sinensis adults in vitro and the survival rate of C. sinensis was analyzed, as well as the expression level and enzymatic activity of CsACAT in different lipid environments. Hypercholesteremia rabbit models were established by feeding with a hyperlipidemic diet and then infected intragastrically with C. sinensis. One and a half months later, the worm burdens and the expression level of CsACAT was detected. RESULTS: CsACAT was confirmed to be a member of the thiolase family and present in the excretory/secretory proteins of C. sinensis. CsACAT was specifically localized at the vitellarium and sub-tegumental muscle layer in adult worms. The mRNA level of CsACAT in eggs was higher than those in adult worms and metacercariae. When adult worms were cultured with higher concentration of lecithin, the expression level and enzyme activity of CsACAT were up-regulated. The survival rate of adult worms was higher than control group. More adult worms were recovered from hypercholesteremia rabbit models. The expression level of CsACAT in these worms was higher than control group. CONCLUSIONS: Our results implied that C. sinensis might sense lipid levels and survive better in the bile environment with higher lipid levels. C. sinensis might modulate the expression and enzymatic activity of CsACAT, an enzyme involved in fatty acid metabolism, for energy or physical requirements to adapt to the host.

Molecular characterization of Clonorchis sinensis secretory myoglobin: delineating its role in anti-oxidative survival.

BACKGROUND: Clonorchiasis is a globally important, neglected food-borne disease caused by Clonorchis sinensis (C. sinensis), and it is highly related to cholangiocarcinoma and hepatocellular carcinoma. Increased molecular evidence has strongly suggested that the adult worm of C. sinensis continuously releases excretory-secretory proteins (ESPs), which play important roles in the parasite-host interactions, to establish successful infection and ensure its own survival. Myoglobin, a hemoprotein, is present in high concentrations in trematodes and ESPs. To further understand the biological function of CsMb and its putative roles in the interactions of C. sinensis with its host, we explored the molecular characterization of CsMb in this paper. METHODS: We expressed CsMb and its mutants in E. coli BL21 and identified its molecular characteristics using bioinformatics analysis and experimental approaches. Reverse transcription PCR analysis was used to measure myoglobin transcripts of C. sinensis with different culture conditions. The peroxidase activity of CsMb was confirmed by spectrophotometry. We co-cultured RAW264.7 cells with recombinant CsMb (rCsMb), and we then measured the production of hydrogen peroxide (H2O2) and nitric oxide (NO) in addition to the mRNA levels of inducible nitric oxide synthase (iNOS), Cu-Zn superoxide dismutase (SOD1) and Mn superoxide dismutase (SOD2) in activated RAW264.7 cells. RESULTS: In the in vitro culture of adult worms, the transcripts of CsMb increased with the increase of oxygen content. Oxidative stress conditions induced by H2O2 increased the levels of CsMb transcripts in a dose-dependent manner. Furthermore, CsMb catalyzed oxidation reactions in the presence of H2O2, and amino acid 34 of CsMb played an essential role in its reaction with H2O2. In addition, CsMb significantly reduced H2O2 and NO levels in LPS-activated macrophages, and CsMb downregulated iNOS and SOD expression in activated macrophages. CONCLUSION: The present study is the first to investigate the peroxidase activity of CsMb. This investigation suggested that C. sinensis may decrease the redox activation of macrophages by CsMb expression to evade host immune responses. These studies contribute to a better understanding of the role of CsMb in the molecular mechanisms involved in ROS detoxification by C. sinensis.

Molecular characterization of severin from Clonorchis sinensis excretory/secretory products and its potential anti-apoptotic role in hepatocarcinoma PLC cells.

BACKGROUND: Clonorchiasis, caused by the infection of Clonorchis sinensis (C. sinensis), is a kind of neglected tropical disease, but it is highly related to cholangiocarcinoma and hepatocellular carcinoma (HCC). It has been well known that the excretory/secretory products of C. sinensis (CsESPs) play key roles in clonorchiasis associated carcinoma. From genome and transcriptome of C. sinensis, we identified one component of CsESPs, severin (Csseverin), which had three putative gelsolin domains. Its homologues are supposed to play a vital role in apoptosis resistance of tumour cell. METHODOLOGY/PRINCIPAL FINDINGS: There was significant similarity in tertiary structures between human gelsolin and Csseverin by bioinformatics analysis. We identified that Csseverin expressed at life stage of adult worm, metacercaria and egg by the method of quantitative real-time PCR and western blotting. Csseverin distributed in vitellarium and intrauterine eggs of adult worm and tegument of metacercaria by immunofluorence assay. We obtained recombinant Csseverin (rCsseverin) and confirmed that rCsseverin could bind with calciumion in circular dichroism spectrum analysis. It was demonstrated that rCsseverin was of the capability of actin binding by gel overlay assay and immunocytochemistry. Both Annexin V/PI assay and mitochondrial membrane potential assay of human hepatocarcinoma cell line PLC showed apoptosis resistance after incubation with different concentrations of rCsseverin. Morphological analysis, apoptosis-associated changes of mitochondrial membrane potential and Annexin V/PI apoptosis assay showed that co-incubation of PLC cells with rCsseverin in vitro led to an inhibition of apoptosis induced by serum-starved for 24 h. CONCLUSIONS/SIGNIFICANCE: Collectively, the molecular properties of Csseverin, a molecule of CsESPs, were characterized in our study. rCsseverin could cause obvious apoptotic inhibition in human HCC cell line. Csseverin might exacerbate the process of HCC patients combined with C. sinensis infection.