Enhancing irrigation water quality efficiently with potassium feldspar-derived adsorbent: Heavy metal detoxification and nutrient augmentation.

The pollution of Pb2+ and Cd2+ in both irrigation water and soil, coupled with the scarcity of vital mineral nutrition, poses a significant hazard to the security and quality of agricultural products. An economical potassium feldspar-derived adsorbent (PFDA) was synthesized using potassium feldspar as the main raw material through ball milling-thermal activation technology to solve this problem. The synthesis process is cost-effective and the resulting adsorbent demonstrates high efficiency in removing Pb2+ and Cd2+ from water. The removal process is endothermic, spontaneous, and stochastic, and follows the quasi-second-order kinetics, intraparticle diffusion, and Langmuir model. The adsorption and elimination of Pb2+ and Cd2+ is largely dependent on monolayer chemical sorption. The maximum removal capacity of PFDA for Pb2+ and Cd2+ at room temperature is 417 and 56.3 mg·g-1, respectively, which is superior to most mineral-based adsorbents. The desorption of Pb2+/Cd2+ on PFDA is highly challenging at pH≥3, whereas PFDA and Pb2+/Cd2+ are recyclable at pH≤0.5. When Pb2+ and Cd2+ coexisted, Pb2+ was preferentially removed by PFDA. In the case of single adsorption, Pb2+ was mainly adsorbed onto PFDA as Pb2SiO4, PbSiO3·xH2O, Pb3SiO5, PbAl2O4, PbAl2SiO6, PbAl2Si2O8, Pb2SO5, and PbSO4, whereas Cd2+ was primarily adsorbed as CdSiO3, Cd2SiO4, and Cd3Al2Si3O12. After the complex adsorption, the main products were PbSiO3·xH2O, PbAl2Si2O8, Pb2SiO4, Pb4Al2Si2O11, Pb5SiO7, PbSO4, CdSiO3, and Cd3Al2Si3O12. The forms of mineral nutrients in single and complex adsorption were different. The main mechanisms by which PFDA removed Pb2+ and Cd2+ were chemical precipitation, complexation, electrostatic attraction, and ion exchange. In irrigation water, the elimination efficiencies of Pb2+ and Cd2+ by PFDA within 10 min were 96.0 % and 70.3 %, respectively, and the concentrations of K+, Si4+, Ca2+, and Mg2+ increased by 14.0 %, 12.4 %, 55.7 %, and 878 %, respectively, within 60 min. PFDA holds great potential to replace costly methods for treating heavy metal pollution and nutrient deficiency in irrigation water, offering a sustainable, cost-effective solution and paving a new way for the comprehensive utilization of potassium feldspar.

The Effect of Two Siderophore-Producing Bacillus Strains on the Growth Promotion of Perennial Ryegrass under Cadmium Stress.

Cadmium (Cd) is a highly toxic and cumulative environmental pollutant. Siderophores are heavy metal chelators with high affinity to heavy metals, such as Cd. Ryegrass (Lolium perenne L.) has a potential remediation capacity for soils contaminated by heavy metals. Consequently, using ryegrass alongside beneficial soil microorganisms that produce siderophores may be an effective means to remediate soils contaminated with Cd. In this study, the Bacillus strains WL1210 and CD303, which were previously isolated from the rhizospheres of Nitraria tangutorum in Wulan and Peganum harmala L. in Dachaidan, Qinghai, China, respectively, both arid and sandy environments, were evaluated for heavy metal pollution mitigation. Our quantitative analyses have discerned that the two bacterial strains possess commendable attributes of phosphorus (P) solubilization and potassium (K) dissolution, coupled with the capacity to produce phytohormones. To assess the heavy metal stress resilience of these strains, they were subjected to a cadmium concentration gradient, revealing their incremental growth despite cadmium presence, indicative of a pronounced tolerance threshold. The subsequent phylogenetic analysis, bolstered by robust genomic data from conserved housekeeping genes, including 16S rDNA, gyr B gene sequencing, as well as dnaK and recA, delineated a species-level phylogenetic tree, thereby confirming the strains as Bacillus atrophaeus. Additionally, we identified the types of iron-carrier-producing strains as catechol (WL1210) and carboxylic acid ferrophilin (CD303). A genomic analysis uncovered functional genes in strain CD303 associated with plant growth and iron carrier biosynthesis, such as fnr and iscA. Ryegrass seed germination assays, alongside morphological and physiological evaluations under diverse heavy metal stress, underscored the strains' potential to enhance ryegrass growth under high cadmium stress when treated with bacterial suspensions. This insight probes the strains' utility in leveraging alpine microbial resources and promoting ryegrass proliferation.

Synthetic Peptides Elicit Humoral Response against Porcine Reproductive and Respiratory Syndrome Virus in Swine.

The aim of this study was to analyze the immunogenic response elicited in swine by two synthetic peptides derived from GP5 to understand the role of lineal B epitopes in the humoral and B-cell-mediated response against the porcine reproductive and respiratory syndrome virus (PRRSV). For inoculation, twenty-one-day-old pigs were allocated into six groups: control, vehicle, vaccinated (Ingelvac-PRRSV, MLV®), non-vaccinated and naturally infected, GP5-B and GP5-B3. At 2 days post-immunization (dpi), the GP5-B3 peptide increased the serum concentrations of cytokines associated with activate adaptive cellular immunity, IL-1ß (1.15 ± 1.15 to 10.17 ± 0.94 pg/mL) and IL-12 (323.8 ± 23.3 to 778.5 ± 58.11 pg/mL), compared to the control group. The concentration of IgGs anti-GP5-B increased in both cases at 21 and 42 dpi compared to that at 0 days (128.3 ± 8.34 ng/mL to 231.9 ± 17.82 and 331 ± 14.86 ng/mL), while IgGs anti-GP5-B3 increased at 21 dpi (105.1 ± 19.06 to 178 ± 15.09 ng/mL) and remained at the same level until 42 dpi. Also, antibody-forming/Plasma B cells (CD2+/CD21-) increased in both cases (9.85 ± 0.7% to 13.67 ± 0.44 for GP5-B and 15.72 ± 1.27% for GP5-B3). Furthermore, primed B cells (CD2-/CD21+) from immunized pigs showed an increase in both cases (9.62 ± 1.5% to 24.51 ± 1.3 for GP5-B and 34 ± 2.39% for GP5-B3) at 42 dpi. Conversely the naïve B cells from immunized pigs decreased compared with the control group (8.84 ± 0.63% to 6.25 ± 0.66 for GP5-B and 5.78 ± 0.48% for GP5-B3). Importantly, both GP5-B and GP5-B3 peptides exhibited immunoreactivity against serum antibodies from the vaccinated group, as well as the non-vaccinated and naturally infected group. In conclusion, GP5-B and GP5-B3 peptides elicited immunogenicity mediated by antigen-specific IgGs and B cell activation.

Development of novel monoclonal antibodies for blocking NF-κB activation induced by CD2v protein in African swine fever virus.

Background: CD2v, a critical outer envelope glycoprotein of the African swine fever virus (ASFV), plays a central role in the hemadsorption phenomenon during ASFV infection and is recognized as an essential immunoprotective protein. Monoclonal antibodies (mAbs) targeting CD2v have demonstrated promise in both diagnosing and combating African swine fever (ASF). The objective of this study was to develop specific monoclonal antibodies against CD2v. Methods: In this investigation, Recombinant CD2v was expressed in eukaryotic cells, and murine mAbs were generated through meticulous screening and hybridoma cloning. Various techniques, including indirect enzyme-linked immunosorbent assay (ELISA), western blotting, immunofluorescence assay (IFA), and bio-layer interferometry (BLI), were employed to characterize the mAbs. Epitope mapping was conducted using truncation mutants and epitope peptide mapping. Results: An optimal antibody pair for a highly sensitive sandwich ELISA was identified, and the antigenic structures recognized by the mAbs were elucidated. Two linear epitopes highly conserved in ASFV genotype II strains, particularly in Chinese endemic strains, were identified, along with a unique glycosylated epitope. Three mAbs, 2B25, 3G25, and 8G1, effectively blocked CD2v-induced NF-κB activation. Conclusions: This study provides valuable insights into the antigenic structure of ASFV CD2v. The mAbs obtained in this study hold great potential for use in the development of ASF diagnostic strategies, and the identified epitopes may contribute to vaccine development against ASFV.

Elucidating the Onset of Cross-Protective Immunity after Intranasal Vaccination with the Attenuated African Swine Fever Vaccine Candidate BA71ΔCD2.

African swine fever (ASF) is a deadly disease of swine currently causing a worldwide pandemic, leading to severe economic consequences for the porcine industry. The control of disease spread is hampered by the limitation of available effective vaccines. Live attenuated vaccines (LAVs) are currently the most advanced vaccine prototypes, providing strong protection against ASF. However, the significant advances achieved using LAVs must be complemented with further studies to analyze vaccine-induced immunity. Here, we characterized the onset of cross-protective immunity triggered by the LAV candidate BA71ΔCD2. Intranasally vaccinated pigs were challenged with the virulent Georgia 2007/1 strain at days 3, 7 and 12 postvaccination. Only the animals vaccinated 12 days before the challenge had effectively controlled infection progression, showing low virus loads, minor clinical signs and a lack of the unbalanced inflammatory response characteristic of severe disease. Contrarily, the animals vaccinated 3 or 7 days before the challenge just showed a minor delay in disease progression. An analysis of the humoral response and whole blood transcriptome signatures demonstrated that the control of infection was associated with the presence of virus-specific IgG and a cytotoxic response before the challenge. These results contribute to our understanding of protective immunity induced by LAV-based vaccines, encouraging their use in emergency responses in ASF-affected areas.

Design and application of Cd2+ polypeptide fluorescent probes based on Aggregation Induced Emission (AIE).

Cadmium is a toxic heavy metal, which is both an environmental pollutant, and a threat to human health. A fluorescent probe was developed to detect Cd2+ selectively, sensitively, and quickly. This study reports the successful development of a polypeptide fluorescent probe TPE-HC (TPE-His-Pro-Gly-Cys) which selectively detects Cd2+ by Aggregation-Induced Emission effect. After fluorescence excitation, Cd2+ can be effectively detected based on the change of fluorescence intensity. The detection limit of Cd2+ in buffer solution was determined to be 151 nM (R2 = 0.9933). This probe exhibits high sensitivity, high cell permeabilit y, and low biological toxicity, and can perform live cell imaging under biological conditions. This study indicates that TPE-HC can detect Cd2+ in biological environments.

A remote-controlled portable workstation for highly sensitive and real-time chemiluminescent detection of cadmium.

The prevention of pollution requires real-time monitoring of cadmium (Cd2+) concentration in the food, as it has a dramatic impact on poultry and can pose a threat to human health. Here, we fabricate a portable workstation integrating a microfluidic chip that facilitates real-time monitoring of Cd2+ levels in real samples by utilizing the Luminol-KMnO4 chemiluminescence (CL) system. Interestingly, Cd2+ can significantly enhance the CL signal, resulting in sensitive detection of Cd2+ in the range of 0-0.18 mg/L with the limit of detection (LOD) of 0.207 µg/L. Furthermore, a remote-controlled unit is integrated into the portable workstation to form a remote-controlled portable workstation (RCPW) performing automated point-of-care testing (POCT) of Cd2+. The as-prepared strategy allows remote control of RCPW to avoid long-distance transportation of samples to achieve real-time target monitoring. Consequently, this system furnishes RCPW for monitoring Cd2+ levels in real samples, thereby holding potential for applications in preventing food pollution.

Biophysical characterization and insights into the oligomeric nature of CD2-associated protein.

INTRODUCTION: Glomerular podocytes are specialized epithelial cells localized to the blood-urine interface of the kidney. Podocyte slit-diaphragm (SD), a size-and-charge-selective junction, is instrumental in blood ultrafiltration and the formation of protein-free urine. The SD consists of macromolecular complexes of several proteins, such as nephrin, podocin, and CD2-associated protein (CD2AP). CD2AP is an adapter protein and is considered to be crucial for the integrity of SD. Mutations in the SD proteins cause nephrotic syndrome (NS), characterized by proteinuria. SD proteins' structural features must be elucidated to understand the mechanism of proteinuria in NS. In this study, we expressed, purified, and biophysically characterized heterologously expressed human CD2AP. METHODS: Codon-optimized human CD2AP was expressed in E. coli Rosetta cells. The recombinant protein was induced with 1 mM IPTG and purified by Ni-NTA affinity chromatography. Analytical size-exclusion chromatography, blue native-PAGE, circular dichroism, and fluorescence spectroscopy were performed to decipher the oligomeric nature, secondary structural content, and tertiary packing of CD2AP. RESULTS: Our analysis revealed that CD2AP adopts a predominantly disordered secondary structure despite exhibiting moderate tertiary packing, characterized by low helical and ß-sheet content. CD2AP readily assembles into homo-oligomers, with octamers and tetramers constituting the primary population. Interestingly, the inherent flexibility of CD2AP's secondary structural elements appears resistant to thermal denaturation. Frameshift mutation (p.K579Efs*7) that leads to loss of the coiled-coil domain promotes aberrant oligomerization of CD2AP through SH3 domains. CONCLUSION: We successfully expressed full-length human CD2AP in a heterologous system, wherein the secondary structure of CD2AP is predominantly disordered. CD2AP can form higher-order oligomers, and the significance of these oligomers and the impact of mutations in the context of size-selective permeability of SD needs further investigation.

Carbon quantum dots as a turn-on fluorescent probe for the sensitive detection of Cd2.

In this study, fluorescent carbon dots were synthesized for the first time using ammonium citrate and glutamic acid as precursors via a one-pot hydrothermal method. The synthesized carbon dots emit blue fluorescence at 436 nm (excited at 320 nm) and demonstrate excellent photobleaching resistance and fluorescence stability in high salt environments. Within the range of 1-25 µM, the fluorescence of CDs gradually increases with the increasing concentration of Cd2+, reaching a limit of detection as low as 13 nM. This phenomenon could be ascribed to the chelation-enhanced fluorescence, a result of Cd2+ forming complexes with the abundant surface functional groups such as CN-, -COOH, -OH, -NH2 in CDs. Furthermore, this turn-on fluorescent probe has been successfully used for the detection of Cd2+ in tap water and lake water, providing an efficient and sensitive method for the analysis of environmental metals.

ß-tricalcium phosphate enhanced biomineralization of Cd2+ and Pb2+ by Sporosarcina ureilytica HJ1 and Sporosarcina pasteurii HJ2.

Microbiologically induced CaCO3 precipitation (MICP) has been proposed as a potential bioremediation method to immobilize contaminating metals. In this study, carbonate mineralizing bacteria HJ1 and HJ2, isolated from heavy metal contaminated soil, was employed for Cd2+ and Pb2+ immobilization with or without ß-tricalcium phosphate addition. Compared with the only treatments amended with strains, the combined application of ß-tricalcium phosphate and HJ1 improved the immobilization rates of Cd and Pb by 1.49 and 1.70 times at 24 h, and the combined application of ß-tricalcium phosphate and HJ2 increased the immobilization rates of Cd and Pb by 1.25 and 1.79 times. The characterization of biomineralization products revealed that Cd2+ and Pb2+ primarily immobilized from the liquid phase as CdCO3 and PbCO3, and the addition of ß-tricalcium phosphate facilitated the formation of Ca4.03Cd0.97(PO4)3(OH) and Pb3(PO4)2. Also, the calcium source was related to the speciation of carbonate precipitation and improved the Cd and Pb remediation efficiency. This research demonstrated the feasibility and effectiveness of MICP combined with ß-tricalcium phosphate in immobilization of Cd and Pb, which will provide a fundamental basis for future applications of MICP to mitigate soil heavy metal pollutions.

Heavy metal ions interactions with G-quadruplex-prone DNA sequences.

The industrial world exposes living organisms to a variety of metal pollutants. Here we investigated whether such elements affect G-rich sequences susceptible to fold into G-quadruplex (GQ) structures. Thermal stability and conformation of these oligoncleotides was studied at various molar ratios of a variety of heavy metal salts using thermal FRET, transition-FRET (t-FRET) and circular dichroism. Metal ions affected the thermal stability of the GQs to different extents; some metals had no effect on Tm while other metals caused small to moderate changes in Tm at 1:1 or 1:10 molar ratio. While most of the metals had no major effect, Al3+, Cd2+, Pb2+, Hg2+ and Zn2+ altered the thermal stability and structural features of the GQs. Some metals such as Pb2+ and Hg2+ exhibit differential interactions with telomere, c-myc and c-kit GQs. Overall, toxic heavy metals affect G-quadruplex stability in a sequence and topology dependent manner. This study provides new insight into how heavy metal exposure may affect gene expression and cellular responses.

Preparation and characterization of high-ash coal slime-based soil amendment as well as investigations of its adsorption performance and mechanisms towards heavy metals in soil.

In this study, high-ash coal slime-based mineral soil amendment (MSA) was prepared via the hydrothermal method using high-ash coal slime as raw material, supplemented with activator calcium oxide and additive KOH solution. After hydrothermal treatment at 230 °C for 5 h, the original crystalline phase (quartz and kaolinite) of the high-ash slime was completely transformed into hydrotalcite zeolite, tobermorite, and silicate of potassium aluminosilicate, which has the largest specific surface area. The adsorption of Pb2+ and Cd2+ was adherent to the kinetic equation of secondary adsorption and Freundlich models, and the removal of Pb2+ and Cd2+ reached up to 362.58 mg g-1 and 64.67 mg g-1. The successive releases of SiO2 and CaO from MSA conformed to the Elovich equation, whereas the releases of SiO2 in Cd-containing environments and CaO in Pb- and Cd-containing environments more closely conformed to the power function; the releases of K2O all conformed to the first-order kinetic equation. The presence of Pb2+ and Cd2+ in the environment promotes the release of potassium and calcium elements with MSA's ion-exchange ability, and attenuates the release of silicon elements. Combining Pb2+ and Cd2+ with silicon resulted in the intolerant precipitation of 3PbO·2SiO2 and Cd2SiO4. The mineral precipitation mechanism is the most important mechanism of MSA in immobilizing heavy metals, accounting for 72.7%-80.5% of the total adsorption. Further contaminated soil immobilization experiments also showed that the application of MSA significantly reduced the bioavailability of soil heavy metals. When the MSA addition amount was 1.6%, the residual state increased by 63.58%. In conclusion, preparing MSA may effectively utilize coal-based solid waste with high added value.

Populus euphratica R2R3-MYB transcription factor RAX2 binds ANN1 promoter to increase cadmium enrichment in Arabidopsis.

The expression of R2R3-MYB transcription factor PeRAX2 increased transiently upon CdCl2 exposure (100 µM, 48 h) in leaves and roots of Populus euphratica. We observed that overexpression of PeRAX2 increased Cd2+ concentration in Arabidopsis root cells and Cd2+ amount in whole plant, which was due to the increased Cd2+ influx into root tips. However, the Cd2+ influx facilitated by PeRAX2 overexpression was substantially reduced by LaCl3 (an inhibitor of Ca2+-channels), suggesting that PeRAX2 could promote the Cd2+ entering through PM Ca2+-permeable channels (CaPCs) in the roots. It is noting that the expression of annexin1 (AtANN1), which mediates the influx of divalent cations through the PM calcium channels, was upregulated by Cd2+ in PeRAX2-transgenic Arabidopsis. Bioinformatic analysis revealed that the AtANN1 promoter (AtANN1-pro) contains four cis-elements for MYB binding. The PeRAX2 interaction with AtANN1-pro was validated by LUC reporter assay, EMSA, and Y1H assay. Our data showed that PeRAX2 binds to the AtANN1 promoter region to regulate gene transcription and that AtANN1 mediates the Cd2+ entry through CaPCs in the PM, leading to a Cd2+ enrichment in transgenic plants. The PeRAX2-stimulated Cd2+ enrichment consequently resulted in high H2O2 production in root cells of transgenic plants. The expression of AtSOD and AtPOD and activities of CAT, SOD, POD increased in the transgenic lines under Cd2+ stress. However, the Cd2+-upregulated expression and activity of antioxidative enzymes were less pronounced in the PeRAX2-overexpressed lines, compared to the wildtype and vector controls. As a result, root length and plant growth were more suppressed by Cd2+ in the transgenic lines. Our data suggest that transcriptional regulation of AtANN1 by PeRAX2 can be utilized to improve Cd2+ enrichment and phytoremediation, although the enriched Cd2+ affected antioxidant defense system and plant growth in the model species.

Nanobodies against African swine fever virus p72 and CD2v proteins as reagents for developing two cELISAs to detect viral antibodies.

African swine fever virus (ASFV) poses a significant threat to the global swine industry. Currently, there are no effective vaccines or treatments available to combat ASFV infection in pigs. The primary means of controlling the spread of the disease is through rapid detection and subsequent elimination of infected pig. Recently, a lower virulent ASFV isolate with a deleted EP402R gene (CD2v-deleted) has been reported in China, which further complicates the control of ASFV infection in pig farms. Furthermore, an EP402R-deleted ASFV variant has been developed as a potential live attenuated vaccine candidate strain. Therefore, it is crucial to develop detection methods that can distinguish wild-type and EP402R-deleted ASFV infections. In this study, two recombinant ASFV-p72 and -CD2v proteins were expressed using a prokaryotic system and used to immunize Bactrian camels. Subsequently, eight nanobodies against ASFV-p72 and ten nanobodies against ASFV-CD2v were screened. Following the production of these nanobodies with horse radish peroxidase (HRP) fusion proteins, the ASFV-p72-Nb2-HRP and ASFV-CD2v-Nb22-HRP fusions were selected for the development of two competitive ELISAs (cELISAs) to detect anti-ASFV antibodies. The two cELISAs exhibited high sensitivity, good specificity, repeatability, and stability. The coincidence rate between the two cELISAs and commercial ELISA kits was 98.6% and 97.6%, respectively. Collectively, the two cELISA for detecting antibodies against ASFV demonstrated ease of operation, a low cost, and a simple production process. The two cELISAs could determine whether pigs were infected with wild-type or CD2v-deleted ASFV, and could play an important role in monitoring ASFV infections in pig farms.

The extracellular polymeric substances (EPS) accumulation of Spirulina platensis responding to Cadmium (Cd2+) exposure.

Spirulina platensis can secrete extracellular polymeric substances (EPS) helping to protect damage from stress environment, such as cadmium (Cd2+) exposure. However, the responding mechanism of S. platensis and the secreted EPS to exposure of Cd2+ is still unclear. This research focuses on the effects of Cd2+ on the composition and structure of the EPS and the response mechanism of EPS secretion from S. platensis for Cd2+ exposure. S. platensis can produce 261.37 mg·g-1 EPS when exposing to 20 mg·L-1 CdCl2, which was 2.5 times higher than the control group. The S. platensis EPS with and without Cd2+ treatment presented similar and stable irregularly fibrous structure. The monosaccharides composition of EPS in Cd2+ treated group are similar with control group but with different monosaccharides molar ratios, especially for Rha, Gal, Glc and Glc-UA. And the Cd2+ treatment resulted in a remarkable decline of humic acid and fulvic acid content. The antioxidant ability of S. platensis EPS increased significantly when exposed to 20 mg·L-1 CdCl2, which could be helpful for S. platensis protecting damage from high concentration of Cd2+. The transcriptome analysis showed that sulfur related metabolic pathways were up-regulated significantly, which promoted the synthesis of sulfur-containing amino acids and the secretion of large amounts of EPS.

High-throughput visualization mutation screening technology to enhance the specificity of CadR based whole-cell cadmium biosensor.

As a heavy metal pollutant, Cd2+ often enters the human body through the food chain causing great harm to human health. Whole-cell biosensor is an emerging technology for rapid on-site detection of heavy metals with the advantages of inexpensive, fast to mass-produce, and strong in anti-interference resistance, but suffering from insatisfactory specificity. In this study, a strategy of Adjacent Site Saturation Mutation (ASSM) was designed to improve the specificity of transcription factor CadR, which acted as the recognition element and determined the specificity of whole cell Cd2+ biosensors. A specific saturated library was constructed using the strategy of adjacent mutation. After two rounds of high-throughput visual screening, a whole-cell biosensor with good response to Cd2+, and with significant weakened Hg2+ interference was obtained. The optimized whole-cell biosensor showed a linear dynamic concentration range from 500 nM to 100 µM, a detection limit of 0.079 µM, and has satisfactory specificity and anti-interference. The ASSM strategy proposed in this study can provide a new method for the application of synthetic biology in food safety detection, indicating the importance of whole-cell biosensors for the detection of heavy metals.

CD2AP is a potential prognostic biomarker of renal clear cell carcinoma.

BACKGROUND: CD2-associated protein (CD2AP) is a podocyte-associated gene and its reduced expression is associated with the development of proteinuria and glomerulosclerosis. However, few studies have focused on the correlation between the expression and prognosis of CD2AP in renal clear cell carcinoma (ccRCC). Therefore, we aimed to assess the regulation of CD2AP expression and prognostic value in ccRCC. METHODS: Multiple databases were employed to examine the expression of CD2AP in ccRCC. RT-qPCR, Western Blot and immunohistochemistry were used to validate CD2AP expression in different cell lines and tissue samples. Kaplan-Meier analysis and ROC curve analysis were performed on the predictive prognostic performance of CD2AP. COX regression was used to construct CD2AP-related prognostic models. The TIMER and TISIDB databases were used to analyze the correlation of tumor-infiltrating immune cells with gene expression, mutations, somatic copy number variation, and immune molecules. Mass spectrometry was used to detect methylation status of the promoter CpG site of CD2AP in multiple cells. RESULTS: We found that CD2AP expression was downregulated in ccRCC and its lower expression level was correlation with worse patient prognosis, higher tumor stage and grade and distant metastasis through analysis of databases, ccRCC cell lines and clinical tissue samples. Moreover, database and mass spectrometry techniques identified and validated cg12968598 hypermethylation as one of the key reasons for the downregulation of CD2AP expression. CD2AP expression was also associated with macrophage and neutrophil infiltration. CONCLUSIONS: Taken together, our results suggest that CD2AP can be used as a diagnostic and prognostic biomarker in ccRCC patients and that DNA hypermethylation plays an important role in reducing CD2AP expression.

An environmentally friendly strategy for reducing the environmental risks of heavy metals adsorbed by kaolinite.

Plenty of heavy metals (HMs) that are adsorbed on clay minerals (such as kaolinite), in addition to low molecular-weight organic acids (such as oxalic acid (OA)) with high activities, are widespread in the natural environment. In the present study, the effects of OA on the environmental behaviors of Pb2+/Cd2+ adsorbed by kaolinite have been investigated. The effectiveness and mechanisms of calcium silicate (CS) and magnesium silicate (MS) in reducing the environmental risks of the HMs have also been studied. The results showed that the releases of Pb2+/Cd2+ increased with an increasing concentration of OA. When different dosages of CS/MS were added to the aging system, a redistribution of HMs took place and the free form of Pb2+/Cd2+ decreased to very low levels. Also, the unextractable Pb2+/Cd2+ increased to high levels. Furthermore, a series of characterizations showed that the released HMs were re-captured by the CS/MS. In addition, the CS immobilized the OA in the solution during the aging process, which also facilitated an immobilization of the carbon element in the environment. In general, the present study has contributed to a further understanding of the transport behaviors of the HMs in natural environments, and of the interactions between CS (or MS), the environmental media, and the heavy metal contaminants. In addition, this study has also provided an eco-friendly strategy for an effective remediation of heavy metal pollution.

Cell type-specific functions of Alzheimer's disease endocytic risk genes.

Endocytosis is a key cellular pathway required for the internalization of cellular nutrients, lipids and receptor-bound cargoes. It is also critical for the recycling of cellular components, cellular trafficking and membrane dynamics. The endocytic pathway has been consistently implicated in Alzheimer's disease (AD) through repeated genome-wide association studies and the existence of rare coding mutations in endocytic genes. BIN1 and PICALM are two of the most significant late-onset AD risk genes after APOE and are both key to clathrin-mediated endocytic biology. Pathological studies also demonstrate that endocytic dysfunction is an early characteristic of late-onset AD, being seen in the prodromal phase of the disease. Different cell types of the brain have specific requirements of the endocytic pathway. Neurons require efficient recycling of synaptic vesicles and microglia use the specialized form of endocytosis-phagocytosis-for their normal function. Therefore, disease-associated changes in endocytic genes will have varied impacts across different cell types, which remains to be fully explored. Given the genetic and pathological evidence for endocytic dysfunction in AD, understanding how such changes and the related cell type-specific vulnerabilities impact normal cellular function and contribute to disease is vital and could present novel therapeutic opportunities. This article is part of a discussion meeting issue 'Understanding the endo-lysosomal network in neurodegeneration'.

Copper oxide nanoparticles alter the uptake and distribution of cadmium through disturbing the ordered structure of the cell wall in Arabidopsis root.

Copper oxide nanoparticles (CuO NPs) influence the uptake of heavy metal ions by plants, but molecular mechanism is still unknown. Here, we proved the mechanism of CuO NPs affecting Cd absorption in Arabidopsis root. 4-d-old seedlings were treated by 10 and 20 mg/L CuO NPs for 3 d, which decreased the contents of cellulose and hemicellulose in roots. Moreover, the contents of some important monosaccharides were altered by CuO NPs, including arabinose, glucose and mannose. Biosynthesis of cellulose and hemicellulose is regulated by cellulose synthase A complexe (CSC) dynamics. The synthesis of tubulin cytoskeleton was inhibited by CuO NPs, which resulted in the decrease of CSCs bidirectional velocities. Furthermore, the arrangement and network of cellulose fibrillar bundles were disrupted by CuO NPs. CuO NPs treatment significantly increased the influx of Cd2+. The accumulation and translocation of Cd were increased by 10 and 20 mg/L CuO NPs treatment. The subcellular distribution of Cd in root cells indicated CuO NPs decrease the enrichment of Cd in cell wall, but increase the enrichment of Cd in soluble fraction and organelle. In light of these findings, we proposed a mechanistic model in which CuO NPs destroy the ordered structure of the cell wall, alter the uptake and distribution of Cd in Arabidopsis.