Transcriptomics analysis of the role of SdiA in desiccation tolerance of Cronobacter sakazakii in powdered infant formula.

The quorum-sensing receptor SdiA is vital for regulating the desiccation tolerance of C. sakazakii, yet the specific mechanism remains elusive. Herein, transcriptomics and phenotypic analysis were employed to explore the response of C. sakazakii wild type (WT) and sdiA knockout strain (ΔsdiA) under drying conditions. Following 20 days of drying in powdered infant formula (PIF), WT exhibited 4 log CFU/g higher survival rates compared to ΔsdiA. Transcriptome revealed similar expression patterns between csrA and sdiA, their interaction was confirmed both by protein-protein interaction analysis and yeast two-hybrid assays. Notably, genes associated with flagellar assembly and chemotaxis (flg, fli, che, mot regulon) showed significantly higher expression levels in WT than in ΔsdiA, indicating a reduced capacity for flagellar synthesis in ΔsdiA, which was consistent with cellular morphology observations. Similarly, genes involved in trehalose biosynthesis (ostAB, treYZS) and uptake (thuEFGK) exhibited similar expression patterns to sdiA, with higher levels of trehalose accumulation observed in WT under desiccation conditions compared to ΔsdiA. Furthermore, WT demonstrated enhanced protein and DNA synthesis capabilities under desiccation stress. Higher expression levels of genes related to oxidative phosphorylation were also noted in WT, ensuring efficient cellular ATP synthesis. This study offers valuable insights into how SdiA influences the desiccation tolerance of C. sakazakii, paving the way for targeted strategies to inhibit and control this bacterium.

Muscle LIM protein of Macrobrachium nipponense (MnMLP) involved in immune and stress response.

The muscle LIM protein (MLP) is a member of the cysteine and glycine-rich protein (CSRP) family, composed of CSRP1, CSRP2 and CSRP3/MLP. MLP is involved in a multitude of functional roles, including cytoskeletal organization, transcriptional regulation, and signal transduction. However, the molecular mechanisms underlying its involvement in immune and stress responses remain to be elucidated. This study identified an MnMLP in the freshwater crustacean Macrobrachium nipponense. The isothermal titration calorimetry assay demonstrated that recombinant MnMLP was capable of coordinating with Zn2+. Upon challenge by Aeromonas veronii or WSSV, and exposure to CdCl2, up-regulation was recorded in the muscle and intestinal tissues, suggesting its involvement in immune and anti-stress responses. MnMLP protein was predominantly expressed in the cytoplasm of the transfected HEK-293T cells, but after treatment with LPS, Cd2+ or H2O2, the MnMLP was observed to be transferred into the nucleus. The comet assay demonstrated that the overexpression of MnMLP could mitigate the DNA damage induced by H2O2 in HEK-293T cells, suggesting the potential involvement of MnMLP in the DNA repair process. These findings suggest that DNA repair may represent a possible mechanism by which MnMLP may be involved in the host's defense against pathogens and stress.

Comprehensive Evaluation of the Physicochemical Attributes, Antioxidant Capacity, and pH-Responsive Behavior of Starch Films Enhanced by Laver Incorporation.

Herein, a new starch film incorporating laver was developed to address issues related to inadequate water resistance and suboptimal preservation quality in food packaging. The integration of laver into starch film formulations offers a compelling avenue for creating biodegradable, active, and smart food packaging. Scanning electron microscope (SEM) analysis revealed that the starch film with a laver concentration of 70% exhibited a uniformly flat microstructure, as expected. Fourier-transform infrared spectroscopy (FTIR) confirmed the presence of intermolecular interactions and hydrogen bonding between the starch and laver. Viscoelastic tests demonstrated the superior film-forming performance of the starch/laver composite films. Moreover, it was found that the most favorable concentration of incorporated laver was 10%. Specifically, the S7-3 film emerged as a promising candidate for food packaging applications, boasting the highest contact angle (CA) value of 114.98 ± 1.28°, the lowest water solubility (WS) value of 15.38%, and a reduced water vapor transmission rate (WVTR) value of 2.52 g/m2 × h. Additionally, the S3-7 film displayed an extraordinary tensile strength of 32.47 MPa, an elongation at break of 19.04%, and a Young's modulus of 606.83 MPa. Furthermore, the starch/laver composite films exhibited outstanding UV-blocking capabilities, exceptional pH-responsive behavior, and significant antioxidant activity, underscoring their potential for packaging applications with laver integration.

Preparation and characterization of peach gum/chitosan polyelectrolyte composite films with dual cross-linking networks for antibacterial packaging.

Peach gum (PG) is a valuable polymeric feedstock for developing eco-friendly, bio-safe, and functional materials. However, PG has limited use in food packaging due to its inferior mechanical and antibacterial properties. To overcome these limitations, we created a dual cross-linked network by introducing chitosan (CS) and glycerol to the PG matrix. Our research discovered that incorporating CS into the PG matrix significantly improved its Young's modulus, from 277.62 to 925.89 MPa, and its tensile strength from 5.96 to 39.94 MPa. Furthermore, the inclusion of glycerol greatly increased the elongation. These enhancements were attributed to the ionic and hydrogen-bonding interactions between the two biopolymers. Additionally, the composite films exhibited strong antibacterial effects, reducing the total number of colonies by 99.2 % and 99.9 % against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus), respectively. The incorporation of CS resulted in more amorphous films, enhancing their stiffness, flexibility, and barrier properties. To assess the practical application of PG/CS composite films, we conducted a comparative analysis between non-packaged strawberries and strawberries packaged with these films. The results demonstrated that the composite polyelectrolyte film extended the shelf life of strawberries better than the non-packaged fruits.

Improving water resistance and mechanical properties of starch-based films by incorporating microcrystalline cellulose in a dynamic network structure.

The widespread use of starch-based films is hindered by inadequate tensile strength and high water sensitivity. To address these limitations, a novel starch film with a dynamic network structure was produced via the dehydration-condensation reaction of N, N'-methylene diacrylamide (MBA) and microcrystalline cellulose (MCC). The improvement in mechanical properties was enhanced by the incorporation of MCC, which was achieved through intermolecular hydrogen bonding and chemical crosslinking. To verify the interactions among MCC, MBA, and starch, x-ray photoelectron spectroscopy (XPS), fourier transform infrared spectroscopy (FTIR), and x-ray diffraction (XRD) were conducted. The results established the predicted interactions. The dynamic network structure of the film reduced the water absorption capacity (WAC) of starch and MCC hydroxyl groups, as confirmed by differential scanning calorimeter (DSC) and dynamic mechanical thermal analysis (DMTA). These analyses showed a restriction in the mobility of starch chains, resulting in a higher glass transition temperature (Tg) of 69.26 °C. The modified starch films exhibited excellent potential for packaging applications, demonstrating a higher contact angle (CA) of 89.63°, the lowest WAC of 4.73 g/g, and the lowest water vapor transmission rate (WVTR) of 13.13 g/m2/d, along with improved mechanical properties and identical light transmittance compared to pure starch films.

CDS-DB, an omnibus for patient-derived gene expression signatures induced by cancer treatment.

Patient-derived gene expression signatures induced by cancer treatment, obtained from paired pre- and post-treatment clinical transcriptomes, can help reveal drug mechanisms of action (MOAs) in cancer patients and understand the molecular response mechanism of tumor sensitivity or resistance. Their integration and reuse may bring new insights. Paired pre- and post-treatment clinical transcriptomic data are rapidly accumulating. However, a lack of systematic collection makes data access, integration, and reuse challenging. We therefore present the Cancer Drug-induced gene expression Signature DataBase (CDS-DB). CDS-DB has collected 78 patient-derived, paired pre- and post-treatment transcriptomic source datasets with uniformly reprocessed expression profiles and manually curated metadata such as drug administration dosage, sampling time and location, and intrinsic drug response status. From these source datasets, 2012 patient-level gene perturbation signatures were obtained, covering 85 therapeutic regimens, 39 cancer subtypes and 3628 patient samples. Besides data browsing, download and search, CDS-DB also supports single signature analysis (including differential gene expression, functional enrichment, tumor microenvironment and correlation analyses), signature comparative analysis and signature connectivity analysis. This provides insights into drug MOA and its heterogeneity in patients, drug resistance mechanisms, drug repositioning and drug (combination) discovery, etc. CDS-DB is available at http://cdsdb.ncpsb.org.cn/.

Modulating the Coordination Environment of Carbon-Dot-Supported Fe Single-Atom Nanozymes for Enhanced Tumor Therapy.

Herein, carbon dot (CD)-supported Fe single-atom nanozymes with high content of pyrrolic N and ultrasmall size (ph-CDs-Fe SAzyme) are fabricated by a phenanthroline-mediated ligand-assisted strategy. Compared with phenanthroline-free nanozymes (CDs-Fe SAzyme), ph-CDs-Fe SAzyme exhibit higher peroxidase (POD)-like activity due to their structure similar to that of ferriporphyrin in natural POD. Aberration-corrected high-angle annular dark field scanning transmission electron microscopy (HAADF-STEM) and X-ray absorption fine structure spectroscopy (XAFS) analyses show that metal Fe is dispersed in ph-CDs-Fe SAzyme as single atoms. Steady-state kinetic studies show that the maximum velocity (Vmax ) and turnover number (kcat ) of H2 O2  homolytic cleavage catalyzed by ph-CDs-Fe SAzyme are 3.0 and 6.2 more than those of the reaction catalyzed by CDs-Fe SAzyme. Density functional theory (DFT) calculations show that the energy barrier of the reaction catalyzed by ph-CDs-Fe SAzyme is lower than that catalyzed by CDs-Fe SAzyme. Antitumor efficacy experiments show that ph-CDs-Fe SAzyme can efficiently inhibit the growth of tumor cells both in vitro and in vivo by synergistic chemodynamic and photothermal effects. Here a new paradigm is provided for the development of efficient antitumor therapeutic approaches based on SAzyme with POD-like activity.

Development and characterization of an active starch-based film as a chlorogenic acid delivery system.

Given its health benefits for the human body, chlorogenic acid (CA) offers promising applications in the food industry. However, the instability and low bioavailability of CA remain to be solved. In this paper, a starch-based film prepared by the homogenization and solution-casting method was used as an effective carrier to alleviate these problems. Homogenization (10-50 MPa) reduced the starch paste viscosity and its particle sizes from 21.64 to 7.68 µm, which promoted the starch recrystallization and induced chemical cross-links between starch-CA, as confirmed by the FTIR result with an appearance of a new CO peak at about 1716 cm-1. Accordingly, the rapidly digestible starch content of the film was reduced to 27.83 % and the CA encapsulation efficiency was increased to 99.08 % (from 65.88 %). As a result, the film system extended CA's release time beyond 4 h and significantly increased the heat-treated CA's antioxidant activity. Besides, the tensile strength and elastic modulus of the film were also improved to 6.29 MPa (from 1.63 MPa) and 160.98 MPa (from 12.02 MPa), respectively, by homogenization. In conclusion, the developed active starch-based film could be used as an edible film for the production of functional food or active food packaging.

How APTMS Acts as a Bridge to Enhance the Compatibility of the Interface between the Hydrophilic Poly(vinyl alcohol) Film and the Hydrophobic Stearic Acid Coating.

The hydrophobic modification of poly(vinyl alcohol) (PVA) film as a biodegradable packaging material has received significant attention in recent research. Despite the use of stearic acid (SA) as a coating for the PVA film, a challenge persists due to the poor compatibility between SA and PVA. This study addressed the aforementioned issue by utilizing (3-aminopropyl)trimethoxysilane (APTMS) as a bridging agent to establish a connection between the hydrophilic PVA film and the hydrophobic SA coating through hydrogen bonding and chemical reactions. First, SEM and EDS analyses confirmed the enhanced interfacial compatibility between the SA coating and the PVA film. Subsequently, the results from 1H NMR, FTIR, and XPS experiments presented evidence of hydrogen bonding and chemical reactions among APTMS, SA, and the PVA film. Interestingly, the PVA-APTMS-SA film demonstrated a contact angle of 120.77°, a water absorption of 7.81%, and a water vapor transmission rate of 8.69 g/m2/h. Furthermore, such a composite film displayed exceptional adhesion performance, requiring detachment stresses of 9.86 ± 0.91 and 6.17 ± 0.75 MPa when tested on glass and marble surfaces, respectively. In conclusion, the PVA-APTMS-SA film exhibited significant potential in extending the freshness of fresh-cut apples, making it a promising eco-friendly packaging material for food preservation.

Study on hydroxypropyl corn starch/alkyl ketene dimer composite film with enhanced water resistance and mechanical properties.

This study aimed to address the limited applicability of starch-based films in food packaging due to their inherent hydrophilicity, by developing a highly hydrophobic and mechanically reinforced film through compositing with alkyl ketene dimer (AKD). The FTIR analysis confirmed the successful introduction of AKD into the starch backbone via esterification by forming a ß-keto ester linkage. Notably, the incorporation of AKD resulted in significant improvements in the modified film (S80A20), by exhibiting a higher water contact angle (WCA) of 128.28° and a reduced water vapor permeability (WVP) to 0.81×10-10 (g m/m2 s Pa). These enhancements were attributed to the inherent low surface energy of AKD and the increased surface roughness caused by AKD recrystallization. Moreover, the mechanical properties of the films were also enhanced due to the chemical crosslinking and intermolecular hydrogen bonding, as supported by the results of relaxation temperatures and molecular dynamics simulations. Considering the environmentally friendly and biodegradable nature of all components, the prepared hydrophobic films will hopefully be applied in food packaging.

E2F family play important roles in tumorigenesis.

Tumors are serious threats to human health. The transcription factors are regarded as the potential targets for tumor treatment. As an important family of transcription factors, E2F family transcription factors (E2Fs) play vital roles in cell proliferation and regulation. However, the expression feature, gene functions, and molecular interactions of E2Fs in tumorigenesis are not clear. In this study, the transcriptome data, mutation data, and protein-protein interaction data of 10 high-incidence tumors in China from the TCGA database were integrated and analyzed to explore the expression, structure, function, mutation, and phylogenetic characteristics of E2Fs. The results showed that E2F1 and E2F7 were regularly upregulated in the tumor samples. Moreover, E2Fs participated in the regulation of the cell cycle, cell aging, and other signaling pathways. As an important regulator, E2F1 interacted with more proteins than other E2Fs. At the same time, the genetic mutation types of E2Fs varied in tumor type and patient sex, of which gene amplification accounts for the largest proportion. Phylogenetic analysis showed that E2Fs were conserved in 41 species, including fruit flies, nematodes, and humans. Meanwhile, E2Fs had a tendency for gene expansion during evolution. In conclusion, this study clarified the expression pattern, mutation characteristics, and evolutionary trend of E2Fs in high-incidence tumors in China, and suggested that E2F family transcription factors could be novel diagnostic markers for tumor diseases. Furthermore, this work can provide a theoretical basis for the development of anti-tumor-targeted drugs.

Long non-coding RNAs regulate heavy metal-induced apoptosis in embryo-derived cells.

Heavy metal pollution has been a worldwide prevalent problem, and particularly a threat to ecosystem integrity and animals' health. Previous studies on the mechanisms of heavy metal toxicity have focused on protein-coding genes, whereas most genomic transcripts are long non-coding RNAs (lncRNAs). Although lncRNAs are known to play important regulatory roles in biological processes, their role in heavy metal stress regulation is still not fully understood. We here developed an insect embryo cell model for studying metal toxicity and the underlying regulatory mechanisms. We performed genome-wide screening and functional characterization of lncRNAs induced by two essential and two non-essential heavy metals in Drosophila embryo-derived S2 cells. We identified 4894 lncRNAs, of which 1410 were novel. Forty-one lncRNAs, together with 328 mRNAs, were induced by all the four heavy metals. LncRNA-mRNA co-expression network and pathway enrichment analysis showed that detoxification metabolism, circadian rhythm, and apoptosis regulation pathways were activated in response to heavy metal stress. LncRNA CR44138 was remarkably upregulated in cells exposed to the four heavy metals and was associated with the apoptosis pathway. Expression interference confirmed that CR44138 aggravated cytotoxicity-induced apoptosis in cells under heavy metals stress. This study highlights the important role of lncRNAs in regulating the cellular response to heavy metals. This study also lays the foundation for discovering the novel regulatory mechanisms and developing diagnostic biomarkers of the toxic effects of heavy metal pollutants on organisms.

Adverse effects of sulfamethoxazole on locomotor behavior and lipid metabolism by inhibiting acetylcholinesterase and lipase in Daphnia magna.

Sulfamethoxazole (SMZ), a kind of sulfonamide antibiotic, is widely used in both human and veterinary medicine. Frequent detection of SMZ in natural aquatic environments has prompted growing attention and posed ecological risks to the ecosystem and human health. In this study, we investigated the ecotoxicological properties of SMZ upon Daphnia magna, and the mechanisms of the deleterious effects of SMZ were attempted to be elucidated by investigating a chain of parameters, including survival, reproduction, growth, locomotor behavior, metabolism, as well as levels of related enzyme activity and gene expression. After a 14-d sub-chronic exposure to SMZ at environmentally relevant concentrations, we observed virtually no lethal effect, weak growth inhibition, significant reproductive damage, evident ingestion rate decline, obvious change in locomotor behavior, and remarkable metabolic disorder. Notably, we identified SMZ as an inhibitor against acetylcholinesterase (AChE)/lipase in D. magna both in vivo and in vitro, which explained the adverse effects of SMZ on locomotor ability and lipid metabolism at the molecular levels. Furthermore, the direct interactions between SMZ and AChE/lipase were confirmed by using fluorescence spectrum and molecular docking. Together, our findings provide a new insight to advance the understanding of the environmental effects of SMZ on freshwater organisms.

Preparation and characterization of a solid dispersion of Hexahydrocolupulone and its application in the preservation of fresh apple juice.

Hops extracts and their derivatives have many important biological activities, among them, excellent antibacterial and antioxidant properties make them a promising food preservative. However, poor water solubility limits their application in the food industry. This work aimed to improve the solubility of Hexahydrocolupulone (HHCL) by preparing solid dispersion (SD) and investigating the application of the obtained products (HHCL-SD) in actual food systems. HHCL-SD was prepared by solvent evaporation with PVPK30 as a carrier. The solubility of HHCL was dramatically increased to 24.72 mg/mL（25 ℃)by preparing HHCL-SD, much higher than that of raw HHCL (0.002 mg/mL). The structure of HHCL-SD and the interaction between HHCL and PVPK30 were analyzed. HHCL-SD was confirmed to have excellent antibacterial and antioxidant activities. Furthermore, the addition of HHCL-SD proved to be beneficial for the sensory, nutritional quality, and microbiological safety of fresh apple juice, hence prolonging its shelf-life.

Single von Willebrand factor C-domain protein confers host defense against white spot syndrome virus by functioning as a pattern recognition receptor in Macrobrachium nipponense.

The single von Willebrand factor C-domain proteins (SVWCs), also known as Vago, are primarily found in arthropods. Their expression was induced by nutritional status, bacterial and viral infections. Despite the prominence of SVWCs in antiviral immunity, the detailed molecular mechanisms remain poorly explained. SVWC has been proposed to elicit antiviral activities through its function as an interferon analog. In contrast, herein, we illustrate that an SVWC homolog from Macrobrachium nipponense (MnSVWC) confers host defense against white spot syndrome virus (WSSV) and covert mortality nodavirus (CMNV) as a pattern recognition receptor (PRR). qRT-PCR analyses demonstrated that the expression of MnSVWC was enhanced upon WSSV infection in all detected tissues, including gills, nerve cords, and hemocytes. Coating WSSV with recombinant MnSVWC (rMnSVWC) promoted the phagocytic activity of hemocytes and subsequent clearance of invasive WSSV from the prawn. On the other hand, the knockdown of MnSVWC with RNAi improved the proliferation ability of WSSV and CMNV in the prawn. Analysis of ELISA and Co-immunoprecipitation (Co-IP) showed that rMnSVWC could bind WSSV by interacting with the vesicle proteins VP26 and VP28. Co-IP analysis verified the interaction between MnSVWC and calmodulin, which implies a vesicle protein-SVWC-calmodulin-clathrin-dependent mechanism underlying the hemocyte-mediated phagocytosis against WSSV. Subsequently, MnSVWC was recognized to activate the expression of transcription factor STAT and an interferon-stimulating gene Viperin, illustrating its involvement in modulating humoral immunity via activation of the JAK/STAT pathway after WSSV infection. These findings indicate that MnSVWC could bind to WSSV as a PRR and participate in the promotion of hemocyte-mediated phagocytosis and the activation of the JAK/STAT pathway in prawns.

Improved hydrophobicity, antibacterial and mechanical properties of polyvinyl alcohol/quaternary chitosan composite films for antibacterial packaging.

Polyvinyl alcohol (PVA) and chitosan (CS) are attractive polymeric feedstocks for developing eco-environmental materials. In this work, a biodegradable and antibacterial film was developed based on PVA blending with different long-chain alkyl and different contents of quaternary chitosan through solution casting, in which quaternary chitosan not only acted as an antibacterial agent but also improved hydrophobicity and mechanical properties. A novel peak appeared at 1470 cm-1 in Transform Infrared Spectroscopy (FTIR) and a new CCl bond spectral peak at 200 eV in X-ray photoelectron spectroscopy (XPS) spectra suggested that CS was successfully modified by quaternary. Besides, the modified films have better antibacterial effects against Escherichia (E. coli) and Staphylococcus (S. aureus) and present stronger antioxidant properties. Optical properties demonstrated that the light transmittance on both UV and visible light showed a decreasing trend with the increase of the quaternary chitosan contents. Whereas the composite films have enhanced hydrophobicity than PVA film. Furthermore, the composite films had higher mechanical properties, in which Young's modulus, tensile strength, and elongation at break were 344.99 MPa, 39.12 MPa, and 507.09 %, respectively. This research demonstrated that the modified composite films could extend the shelf of life on antibacterial packaging.

Ecological succession and functional characteristics of lactic acid bacteria in traditional fermented foods.

Fermented foods are important parts of traditional food culture with a long history worldwide. Abundant nutritional materials and open fermentation contribute to the diversity of microorganisms, resulting in unique product quality and flavor. Lactic acid bacteria (LAB), as important part of traditional fermented foods, play a decisive role in the quality and safety of fermented foods. Reproduction and metabolic of microorganisms drive the food fermentation, and microbial interaction plays a major role in the fermentation process. Nowadays, LAB have attracted considerable interest due to their potentialities to add functional properties to certain foods or as supplements along with the research of gut microbiome. This review focuses on the characteristics of diversity and variability of LAB in traditional fermented foods, and describes the principal mechanisms involved in the flavor formation dominated by LAB. Moreover, microbial interactions and their mechanisms in fermented foods are presented. They provide a theoretical basis for exploiting LAB in fermented foods and improving the quality of traditional fermented foods. The traditional fermented food industry should face the challenge of equipment automation, green manufacturing, and quality control and safety in the production.

Peptidoglycan recognition protein SC (PGRP-SC) shapes gut microbiota richness, diversity and composition by modulating immunity in the house fly Musca domestica.

The gastrointestinal tract of all animals, including insects, is colonized by a remarkable array of microorganisms which are referred to collectively as the gut microbiota. The hosts establish mutually beneficial interactions with the gut microbiota. However, the mechanisms shaping these interactions remain to be better understood. Here, we investigated the roles of Musca domestica peptidoglycan recognition protein SC (MdPGRP-SC), a secreted pattern recognition receptor, in shaping the gut microbial community structure by using biochemical and high-throughput sequencing approaches. The recombinant MdPGRP-SC (rMdPGRP-SC) could strongly bind various pathogen-associated molecular patterns (PAMPs) including peptidoglycan, lipopolysaccharide and D-galactose, and exhibited mild affinity to ß-1, 3-glucan and D-mannose. Meanwhile, rMdPGRP-SC could also bind different kinds of microorganisms, including gram-positive bacteria (Bacillus subtilis and Staphylococcus aureus), gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa) and yeast (Pichia pastoris). rMdPGRP-SC also exhibited weak antibacterial activity against Bacillus subtilis. Knockdown of MdPGRP-SC by RNAi reduced the persistence of ingested E. coli and a load of indigenous microbiota in the larval gut significantly. In addition, depleted MdPGRP-SC also altered the gut microbiota composition and led to increased ratios of Gram-negative bacteria. We hypothesize that MdPGRP-SC is involved in maintaining gut homeostasis by modulating the immune intensity of the gut through multiple mechanisms, including degrading or neutralizing various PAMPs and selectively suppressing the growth of some bacteria. Considering the functional conservation of the peptidoglycan recognition protein (PGRP) family in insects, the catalytic PGRPs might be promising candidate targets not only for pest and vector control but also for the treatment of bacterial infection in insect farming.

Protective effects of small heat shock proteins in Daphnia magna against heavy metal exposure.

Daphnia magna is one of the most commonly used model organisms to assess toxicity of heavy metal and other xenobiotics. However, the lack of knowledge about important stress-resistant molecules limits our understanding of the alteration of phenotypic and physiological traits of D. magna upon stress exposures. In this study, we focused on a chaperone family of small heat shock protein (sHSP) that has been found in archaea, bacteria and eukaryotes and plays an important role in stress tolerance. A total of eleven sHSP genes (termed DmsHSP1 - DmsHSP11) were identified from the D. magna genome, whose expression profiles during exposure to heavy metal (Cd2+, Cu2+ and Zn2+) and a few other potential pollutants were evaluated via qRT-PCR and RNA-Seq analysis. The results highlighted the predominant role of DmsHSP1 with the highest basal expression level in adults and robust upregulation upon exposure to heavy metals (Cu2+ > Cd2+ > Zn2+). In vivo, recombinant protein rDmsHSP1-21 and rDmsHSP11-12.8 could not only prevent model substrates agglutination induced by heavy metals or reducer dithiotreitol (DTT), but also protect tissue proteins and enzymes from denaturation and inactivation caused by heavy metals or high temperature. Ectopically expression of DmsHSP1-21 or DmsHSP11-12.8 in E. coli conferred host enhanced resistance against various abiotic stresses including Cd2+, Cu2+ and phenazine methosulfate (PMS). Knockdown of DmsHSP1-21 by RNAi, but not for DmsHSP11-12.8, significantly increased the vulnerability of D. magna to heavy metal exposure. Our work provides systematic information on the evolution and function of sHSPs in D. magna and leads to important insights into the mechanisms by which D. magna survive in adverse environments.