Induction of transgenerational toxicity is associated with the activated germline insulin signals in nematodes exposed to nanoplastic at predicted environmental concentrations.

Exposure to nanoplastics can induce toxicity on organisms at both parental generation (P0-G) and the offspring. However, the underlying mechanism remains unknown. Using Caenorhabditis elegans as a model organism, exposure to 20-nm polystyrene nanoparticle (PS-NP) (1-100 µg/L) upregulated the expressions of insulin ligands (INS-39, INS-3, and DAF-28), and this increase could be further detected in the offspring after PS-NP exposure. Germline ins-39, ins-3, and daf-28 RNAi induced resistance to transgenerational toxicity of PS-NP, indicating that increase in expression of these three insulin ligands mediated induction of transgenerational toxicity. These three insulin ligands transgenerationally activated function of insulin receptor DAF-2 to control transgenerational toxicity of PS-NP. Exposure to 1-100 µg/L PS-NP further upregulated DAF-2, AGE-1, and AKT-1 expressions and downregulated DAF-16 expression. During transgenerational toxicity control, DAF-16/AKT-1/AGE-1 was identified as downstream signaling cascade of DAF-2. Moreover, transcriptional factor DAF-16 activated two downstream targets of HSP-6 (a mitochondrial UPR marker) and SOD-3 (a mitochondrial SOD) to modulate transgenerational toxicity of PS-NP. Our findings indicate a crucial link between activation of insulin signaling and induction of transgenerational toxicity of nanoplastics at low concentrations in organisms.

In Situ Characterization of Dehydration during Ion Transport in Polymeric Nanochannels.

The transport of hydrated ions across nanochannels is central to biological systems and membrane-based applications, yet little is known about their hydrated structure during transport due to the absence of in situ characterization techniques. Herein, we report experimentally resolved ion dehydration during transmembrane transport using modified in situ liquid ToF-SIMS in combination with MD simulations for a mechanistic reasoning. Notably, complete dehydration was not necessary for transport to occur across membranes with sub-nanometer pores. Partial shedding of water molecules from ion solvation shells, observed as a decrease in the average hydration number, allowed the alkali-metal ions studied here (lithium, sodium, and potassium) to permeate membranes with pores smaller than their solvated size. We find that ions generally cannot hold more than two water molecules during this sterically limited transport. In nanopores larger than the size of the solvation shell, we show that ionic mobility governs the ion hydration number distribution. Viscous effects, such as interactions with carboxyl groups inside the membrane, preferentially hinder the transport of the mono- and dihydrates. Our novel technique for studying ion solvation in situ represents a significant technological leap for the nanofluidics field and may enable important advances in ion separation, biosensing, and battery applications.

Three-Dimensional Analysis of the Natural-Organic-Matter Distribution in the Cake Layer to Precisely Reveal Ultrafiltration Fouling Mechanisms.

Cake layer formation is the dominant ultrafiltration membrane fouling mechanism after long-term operation. However, precisely analyzing the cake-layer structure still remains a challenge due to its thinness (micro/nano scale). Herein, based on the excellent depth-resolution and foulant-discrimination of time-of-flight secondary ion mass spectrometry, a three-dimensional analysis of the cake-layer structure caused by natural organic matter was achieved at lower nanoscale for the first time. When humic substances or polysaccharides coexisted with proteins separately, a homogeneous cake layer was formed due to their interactions. Consequently, membrane fouling resistances induced by proteins were reduced by humic substances or polysaccharides, leading to a high flux. However, when humic substances and polysaccharides coexisted, a sandwich-like cake layer was formed owing to the asynchronous deposition based on molecular dynamics simulations. As a result, membrane fouling resistances were superimposed, and the flux was low. Furthermore, it is interesting that cake-layer structures were relatively stable under common UF operating conditions (i.e., concentration and stirring). These findings better elucidate membrane fouling mechanisms of different natural-organic-matter mixtures. Moreover, it is demonstrated that membrane fouling seems lower with a more homogeneous cake layer, and humic substances or polysaccharides play a critical role. Therefore, regulating the cake-layer structure by feed pretreatment scientifically based on proven mechanisms should be an efficient membrane-fouling-control strategy.

Effect of Capecitabine Maintenance Therapy Using Lower Dosage and Higher Frequency vs Observation on Disease-Free Survival Among Patients With Early-Stage Triple-Negative Breast Cancer Who Had Received Standard Treatment: The SYSUCC-001 Randomized Clinical Trial.

Importance: Among all subtypes of breast cancer, triple-negative breast cancer has a relatively high relapse rate and poor outcome after standard treatment. Effective strategies to reduce the risk of relapse and death are needed. Objective: To evaluate the efficacy and adverse effects of low-dose capecitabine maintenance after standard adjuvant chemotherapy in early-stage triple-negative breast cancer. Design, Setting, and Participants: Randomized clinical trial conducted at 13 academic centers and clinical sites in China from April 2010 to December 2016 and final date of follow-up was April 30, 2020. Patients (n = 443) had early-stage triple-negative breast cancer and had completed standard adjuvant chemotherapy. Interventions: Eligible patients were randomized 1:1 to receive capecitabine (n = 222) at a dose of 650 mg/m2 twice a day by mouth for 1 year without interruption or to observation (n = 221) after completion of standard adjuvant chemotherapy. Main Outcomes and Measures: The primary end point was disease-free survival. Secondary end points included distant disease-free survival, overall survival, locoregional recurrence-free survival, and adverse events. Results: Among 443 women who were randomized, 434 were included in the full analysis set (mean [SD] age, 46 [9.9] years; T1/T2 stage, 93.1%; node-negative, 61.8%) (98.0% completed the trial). After a median follow-up of 61 months (interquartile range, 44-82), 94 events were observed, including 38 events (37 recurrences and 32 deaths) in the capecitabine group and 56 events (56 recurrences and 40 deaths) in the observation group. The estimated 5-year disease-free survival was 82.8% in the capecitabine group and 73.0% in the observation group (hazard ratio [HR] for risk of recurrence or death, 0.64 [95% CI, 0.42-0.95]; P = .03). In the capecitabine group vs the observation group, the estimated 5-year distant disease-free survival was 85.8% vs 75.8% (HR for risk of distant metastasis or death, 0.60 [95% CI, 0.38-0.92]; P = .02), the estimated 5-year overall survival was 85.5% vs 81.3% (HR for risk of death, 0.75 [95% CI, 0.47-1.19]; P = .22), and the estimated 5-year locoregional recurrence-free survival was 85.0% vs 80.8% (HR for risk of locoregional recurrence or death, 0.72 [95% CI, 0.46-1.13]; P = .15). The most common capecitabine-related adverse event was hand-foot syndrome (45.2%), with 7.7% of patients experiencing a grade 3 event. Conclusions and Relevance: Among women with early-stage triple-negative breast cancer who received standard adjuvant treatment, low-dose capecitabine maintenance therapy for 1 year, compared with observation, resulted in significantly improved 5-year disease-free survival. Trial Registration: ClinicalTrials.gov Identifier: NCT01112826.

Comparative Analysis of the Glycosylation Profiles of Membrane-Anchored HIV-1 Envelope Glycoprotein Trimers and Soluble gp140.

UNLABELLED: The human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein (Env) trimer, which consists of the gp120 and gp41 subunits, is the focus of multiple strategies for vaccine development. Extensive Env glycosylation provides HIV-1 with protection from the immune system, yet the glycans are also essential components of binding epitopes for numerous broadly neutralizing antibodies. Recent studies have shown that when Env is isolated from virions, its glycosylation profile differs significantly from that of soluble forms of Env (gp120 or gp140) predominantly used in vaccine discovery research. Here we show that exogenous membrane-anchored Envs, which can be produced in large quantities in mammalian cells, also display a virion-like glycan profile, where the glycoprotein is extensively decorated with high-mannose glycans. Additionally, because we characterized the glycosylation with a high-fidelity profiling method, glycopeptide analysis, an unprecedented level of molecular detail regarding membrane Env glycosylation and its heterogeneity is presented. Each glycosylation site was characterized individually, with about 500 glycoforms characterized per Env protein. While many of the sites contain exclusively high-mannose glycans, others retain complex glycans, resulting in a glycan profile that cannot currently be mimicked on soluble gp120 or gp140 preparations. These site-level studies are important for understanding antibody-glycan interactions on native Env trimers. Additionally, we report a newly observed O-linked glycosylation site, T606, and we show that the full O-linked glycosylation profile of membrane-associated Env is similar to that of soluble gp140. These findings provide new insight into Env glycosylation and clarify key molecular-level differences between membrane-anchored Env and soluble gp140. IMPORTANCE: A vaccine that protects against human immunodeficiency virus type 1 (HIV-1) infection should elicit antibodies that bind to the surface envelope glycoproteins on the membrane of the virus. The envelope glycoproteins have an extensive coat of carbohydrates (glycans), some of which are recognized by virus-neutralizing antibodies and some of which protect the virus from neutralizing antibodies. We found that the HIV-1 membrane envelope glycoproteins have a unique pattern of carbohydrates, with many high-mannose glycans and also, in some places, complex glycans. This pattern was very different from the carbohydrate profile seen for a more easily produced soluble version of the envelope glycoprotein. Our results provide a detailed characterization of the glycans on the natural membrane envelope glycoproteins of HIV-1, a carbohydrate profile that would be desirable to mimic with a vaccine.

Vaccine-induced HIV-1 envelope gp120 constant region 1-specific antibodies expose a CD4-inducible epitope and block the interaction of HIV-1 gp140 with galactosylceramide.

UNLABELLED: Mucosal epithelial cell surface galactosylceramide (Galcer) has been postulated to be a receptor for HIV-1 envelope (Env) interactions with mucosal epithelial cells. Disruption of the HIV-1 Env interaction with such alternate receptors could be one strategy to prevent HIV-1 entry through the mucosal barrier. To study antibody modulation of HIV-1 Env-Galcer interactions, we used Galcer-containing liposomes to assess whether natural- and vaccine-induced monoclonal antibodies can block HIV-1 Env binding to Galcer. HIV-1 Env gp140 proteins bound to Galcer liposomes with Kds (dissociation constants) in the nanomolar range. Several HIV-1 ALVAC/AIDSVAX vaccinee-derived monoclonal antibodies (MAbs) specific for the gp120 first constant (C1) region blocked Galcer binding of a transmitted/founder HIV-1 Env gp140. Among the C1-specific MAbs that showed Galcer blocking, the antibody-dependent cellular cytotoxicity-mediating CH38 IgG and its natural IgA isotype were the most potent blocking antibodies. C1-specific IgG monoclonal antibodies that blocked Env binding to Galcer induced upregulation of the gp120 CD4-inducible (CD4i) epitope bound by MAb 17B, demonstrating that a conformational change in gp120 may be required for Galcer blocking. However, the MAb 17B itself did not block Env-Galcer binding, suggesting that the C1 antibody-induced gp120 conformational changes resulted in alteration in a Galcer binding site distant from the CD4i 17B MAb binding site. IMPORTANCE: Galactosyl ceramide, a glycosphingolipid, has been postulated to be a receptor for the HIV-1 envelope glycoprotein (Env) interaction with mucosal epithelial cells. Here, we have mimicked this interaction by using an artificial membrane containing synthetic Galcer and recombinant HIV-1 Env proteins to identify antibodies that would block the HIV-1 Env-Galcer interaction. Our study revealed that a class of vaccine-induced human antibodies potently blocks HIV-1 Env-Galcer binding by perturbing the HIV-1 Env conformation.

Polyethylene nanoparticles at environmentally relevant concentrations enhances neurotoxicity and accumulation of 6-PPD quinone in Caenorhabditis elegans.

The exposure risk of 6-PPD quinone (6-PPDQ) has aroused increasing concern. In the natural environment, 6-PPDQ could interact with other pollutants, posing more severe environmental problems and toxicity to organisms. We here examined the effect of polyethylene nanoplastic (PE-NP) on 6-PPDQ neurotoxicity and the underling mechanisms in Caenorhabditis elegans. In nematodes, PE-NP (1 and 10 µg/L) decreased locomotion behavior, but did not affect development of D-type neurons. Exposure to PE-NP (1 and 10 µg/L) strengthened neurotoxicity of 6-PPDQ (10 µg/L) on the aspect of locomotion and neurodegeneration induction of D-type motor neurons. Exposure to PE-NPs (10 µg/L) caused increase in expressions of mec-4, asp-3, and asp-4 governing neurodegeneration in 10 µg/L 6-PPDQ exposed nematodes. Moreover, exposure to PE-NP (10 µg/L) increased expression of some neuronal genes (daf-7, dbl-1, jnk-1, and mpk-1) in 6-PPDQ exposed nematodes, and RNAi of these genes resulted in susceptibility to neurotoxicity of PE-NP and 6-PPDQ. 6-PPDQ could be adsorbed by PE-NPs, and resuspension of PE-NP and 6-PPDQ after adsorption equilibrium exhibited similar neurotoxicity to co-exposure of PE-NP and 6-PPDQ. In addition, exposure to PE-NP (1 and 10 µg/L) increased 6-PPDQ accumulation in body of nematodes and increased defecation cycle length in 6-PPDQ exposed nematodes. Therefore, 6-PPDQ could be adsorbed on nanoplastics (such as PE-NPs) and enhance both neurotoxicity and accumulation of 6-PPDQ in organisms.

Comparison of reproductive toxicity between pristine and aged polylactic acid microplastics in Caenorhabditis elegans.

Caenorhabditis elegans was employed as model to compare reproductive toxicity between pristine and aged polylactic acid microplastics (PLA-MPs). Aged PLA-MPs induced by UV irradiation showed degradation reflected by decrease in size and alteration in morphological surface. Aged PLA-MPs also exhibited some certain changes of chemical properties compared to pristine PLA-MP. Compared with pristine PLA-MPs, more severe toxicity on reproductive capacity and gonad development was detected in 1-100 µg/L aged PLA-MPs. Meanwhile, aged PLA-MPs caused more severe enhancement in germline apoptosis and alterations in expressions of ced-9, ced-4, ced-3, and egl-1 governing cell apoptosis. In addition, aged PLA-MPs resulted in more severe increase in expression of DNA damage related genes (cep-1, mrt-2, hus-1, and clk-2) compared to pristine PLA-MPs, and the alterations in expression of ced-9, ced-4, ced-3, and egl-1 in pristine and aged PLA-MPs could be reversed by RNAi of cep-1, mrt-2, hus-1, and clk-2. Besides this, enhanced germline apoptosis in pristine and aged PLA-MPs exposed animals was also suppressed by RNAi of cep-1, mrt-2, hus-1, and clk-2. Therefore, our results suggested the more severe exposure risk of aged PLA-MPs than pristine PLA-MPs in causing reproductive toxicity, which was associated with the changed physicochemical properties and DNA damage induced germline apoptosis.

Nanoplastic at environmentally relevant concentrations induces toxicity across multiple generations associated with inhibition in germline G protein-coupled receptor CED-1 in Caenorhabditis elegans.

Nanoplastics at environmentally relevant concentrations (ERCs) could cause transgenerational toxicity on organisms. Caenorhabditis elegans is an important model for the study of transgenerational toxicology of pollutants. Nevertheless, the underlying mechanisms for the control of transgenerational nanoplastic toxicity by germline signals remain largely unclear. In C. elegans, exposure to 1-100 µg/L polystyrene nanoparticle (PS-NP) decreased expression of germline ced-1 encoding a G protein-coupled receptor at parental generation (P0-G). After PS-NP exposure at P0-G, transgenerational decrease in germline ced-1 expression could be detected. Meanwhile, the susceptibility to transgenerational PS-NP toxicity was observed in ced-1(RNAi) animals. After PS-NP exposure at P0-G, germline RNAi of ced-1 increased expressions of met-2 and set-6 encoding histone methylation transferases. The susceptibility of ced-1(RNAi) to transgenerational PS-NP toxicity could be inhibited by RNAi of met-2 and set-6. Moreover, in PS-NP exposed met-2(RNAi) and set-6(RNAi) nematodes, expressions of ins-39, wrt-3, and/or efn-3 encoding secreted ligands were decreased. Therefore, our results demonstrated that inhibition in germline CED-1 mediated the toxicity induction of nanoplastics at ERCs across multiple generations in nematodes.

Garlic-Derived Exosome-like Nanovesicles-Based Wound Dressing for Staphylococcus aureus Infection Visualization and Treatment.

Garlic-derived exosome-like nanovesicles (GELNs) could function in interspecies communication and may serve as natural therapeutics to regulate the inflammatory response or as nanocarriers to efficiently deliver specific drugs. Staphylococcus aureus (S. aureus) is able to hide within host cells to evade immune clearance and antibiotics, leading to life-threatening infections. On-site detection and efficient treatment of intracellular S. aureus infection in wounds remain challenging. Herein, we report a thermosensitive, injectable, visible GELNs-based wound dressing, Van@GELNs/F127 hydrogel (gel Van@GELNs), which is H2O2-responsive and can slowly release vancomycin into host cells forS. aureus infection visualization and treatment in wounds. GELNs show inherent antibacterial activity, which is significantly enhanced after loading vancomycin. Both GELNs and Van@GELNs have the ability to be internalized by cells, so Van@GELNs are more effective than free vancomycin in killing S. aureus in RAW 264.7 macrophages. When applied to an S. aureus-infected wound on a mouse, the colorless HRP&ABTS/Van@GELNs/F127 solution immediately changes to a green hydrogel and shows better therapeutic effect than vancomycin. Thus, direct visualization by the naked eye and effective treatment of S. aureus infection in wounds are achieved by gel Van@GELNs. We anticipate gel Van@GELNs be applied for the theranostics of S. aureus infection diseases in the clinic in the near future.

Tire-rubber related pollutant 6-PPD quinone: A review of its transformation, environmental distribution, bioavailability, and toxicity.

The antioxidant 6-PPD has been widely used to prevent cracking and thermal oxidative degradation and to extend the service life of tire rubber. 6-PPD quinone (6-PPDQ) is formed via the reaction of 6-PPD with O3. Due to its acute lethality in coho salmon, 6-PPDQ has become an emerging pollutant of increasing concern. In this review, we provide a critical overview of the generation, environmental distribution, bioavailability, and potential toxicity of 6-PPDQ. The transformation pathways from 6-PPD to 6-PPDQ include the N-1,3-dimethylbutyl-N-phenyl quinone diamine (QDI), intermediate phenol, and semiquinone radical pathways. 6-PPDQ has been frequently detected in water, dust, air particles, soil, and sediments, indicating its large-scale and potentially global pollution trend. 6-PPDQ is bioavailable to both aquatic animals and mammals and acute exposure to 6-PPDQ can be lethal to some organisms. Exposure to 6-PPDQ at environmentally relevant concentrations could induce several types of toxicity, including neurotoxicity, intestinal toxicity, and reproductive toxicity. This review also identifies and discusses knowledge gaps and research needs for the study of 6-PPDQ. This review facilitates a better understanding of the environmental occurrence and exposure risk of 6-PPDQ.

Paeoniflorin attenuates polystyrene nanoparticle-induced reduction in reproductive capacity and increase in germline apoptosis through suppressing DNA damage checkpoints in Caenorhabditis elegans.

Due to high sensitivity to environmental exposures, Caenorhabditis elegans is helpful for toxicity evaluation and toxicological study of pollutants. Using this animal model, we investigated the reproductive toxicity of 20 nm polystyrene nanoparticle (PS-NP) in the range of µg/L and the following pharmacological intervention of paeoniflorin. After exposure from L1-larvae to young adults, 10-100 µg/L PS-NP could cause the reduction in reproductive capacity reflected by the endpoints of brood size and number of fertilized eggs in uterus. Meanwhile, the enhancements in germline apoptosis analyzed by AO staining and germline DNA damage as shown by alteration in HUS-1::GFP signals were detected in 10-100 µg/L PS-NP exposed nematodes, suggesting the role of DNA damage-induced germline apoptosis in mediating PS-NP toxicity on reproductive capacity. Following the exposure to 100 µg/L PS-NP, posttreatment with 25-100 mg/L paeoniflorin increased the reproductive capacity and inhibited both germline apoptosis and DNA damage. In addition, in 100 µg/L PS-NP exposed nematodes, treatment with 100 mg/L paeoniflorin modulated the expressions of genes governing germline apoptosis as indicated by the decrease in ced-3, ced-4, an egl-1 expressions and the increase in ced-9 expression. After exposure to 100 µg/L PS-NP, treatment with 100 mg/L paeoniflorin also decreased expressions of genes (cep-1, clk-2, hus-1, and mrt-2) governing germline DNA damage. Molecular docking analysis further demonstrated the binding potential of paeoniflorin with three DNA damage checkpoints (CLK-2, HUS-1, and MRT-2). Therefore, our data suggested the toxicity of PS-NP in the range of µg/L on reproductive capacity after exposure from L1-larvae to young adults, which was associated with the enhancement in DNA damage-induced germline apoptosis. More importantly, the PS-NP-induced reproductive toxicity on nematodes could be inhibited by the following paeoniflorin treatment.

Activation of FGF signal in germline mediates transgenerational toxicity of polystyrene nanoparticles at predicted environmental concentrations in Caenorhabditis elegans.

Nanoplastics in the environment could cause the ecological and health risks. Recently, the transgenerational toxicity of nanoplastic has been observed in different animal models. In this study, using Caenorhabditis elegans as an animal model, we aimed to examine the role of alteration in germline fibroblast growth factor (FGF) signal in mediating the transgenerational toxicity of polystyrene nanoparticle (PS-NP). Exposure to 1-100 µg/L PS-NP (20 nm) induced transgenerational increase in expressions of germline FGF ligand/EGL-17 and LRP-1 governing FGF secretion. Germline RNAi of egl-17 and lrp-1 resulted in resistance to transgenerational PS-NP toxicity, indicating the requirement of FGF ligand activation and secretion in formation of transgenerational PS-NP toxicity. Germline overexpression of EGL-17 increased expression of FGF receptor/EGL-15 in the offspring, and RNAi of egl-15 at F1 generation (F1-G) inhibited transgenerational toxicity of PS-NP exposed animals overexpressing germline EGL-17. EGL-15 functions in both the intestine and the neurons to control transgenerational PS-NP toxicity. Intestinal EGL-15 acted upstream of DAF-16 and BAR-1, and neuronal EGL-15 functioned upstream of MPK-1 to control PS-NP toxicity. Our results suggested the important role of activation in germline FGF signal in mediating the induction of transgenerational toxicity in organisms exposed to nanoplastics in the range of µg/L.

Polylactic acid microparticles in the range of µg/L reduce reproductive capacity by affecting the gonad development and the germline apoptosis in Caenorhabditis elegans.

Polylactic acid (PLA) accounts for approximately 45% of the global market of biodegradable plastics. Using Caenorhabditis elegans as an animal model, we examined the effect of long-term exposure to PLA microplastic (MP) on reproductive capacity and the underlying mechanism. Brood size, number of fertilized eggs in uterus, and number of hatched eggs were significantly reduced by exposure to 10 and 100 µg/L PLA MP. Number of mitotic cells per gonad, area of gonad arm, and length of gonad arm were further significantly decreased by exposure to 10 and 100 µg/L PLA MP. In addition, exposure to 10 and 100 µg/L PLA MP enhanced germline apoptosis in the gonad. Accompanied with the enhancement in germline apoptosis, exposure to 10 and 100 µg/L PLA MP decreased expression of ced-9 and increased expressions of ced-3, ced-4, and egl-1. Moreover, the induction of germline apoptosis in PLA MP exposed nematodes was suppressed by RNAi of ced-3, ced-4, and egl-1, and strengthened by RNAi of ced-9. Meanwhile, we did not detect the obvious effect of leachate of 10 and 100 µg/L PLA MPs on reproductive capacity, gonad development, germline apoptosis, and expression of apoptosis related genes. Therefore, exposure to 10 and 100 µg/L PLA MPs potentially reduces the reproductive capacity by influencing the gonad development and enhancing the germline apoptosis in nematodes.

Long-term exposure to tire-derived 6-PPD quinone causes intestinal toxicity by affecting functional state of intestinal barrier in Caenorhabditis elegans.

2-((4-Methylpentan-2-yl)amino)-5-(phenylamino)cyclohexa-2,5-diene-1,4-dione (6-PPDQ) is the ozonation product of 6-PPD, a commonly used tire preservative. Although the 6-PPDQ has been frequently detected in different environmental ecosystems, its long-term effects on organisms remain still largely unknown. We here used Caenorhabditis elegans as an experimental animal to investigate the toxic effect of prolonged exposure to 6-PPDQ (0.1-100 µg/L). After the exposure, we found that 100 µg/L 6-PPDQ caused the lethality. We further selected concentrations of 0.1-10 µg/L to examine the possible intestinal toxicity induced by 6-PPDQ. Although 0.1-10 µg/L 6-PPDQ could not influence intestinal morphology, the intestinal permeability was significantly enhanced by 1-10 µg/L 6-PPDQ as indicated by erioglaucine disodium staining. In addition, the expression of intestinal fatty acid transporter ACS-22 governing functional state of intestinal barrier was decreased by exposure to 1-10 µg/L 6-PPDQ. Meanwhile, intestinal reactive oxygen species (ROS) production was induced by 0.1-10 µg/L 6-PPDQ and lipofuscin accumulation reflected by intestinal autofluorescence was activated by 1-10 µg/L 6-PPDQ. Accompanied with activation of intestinal oxidative stress, expressions of some anti-oxidation related genes (ctl-2, sod-2, sod-3, and sod-4) were significantly increased by 0.1-10 µg/L 6-PPDQ. Moreover, intestinal RNAi of acs-22 strengthened the susceptibility of nematodes to intestinal toxicity of 6-PPDQ. Therefore, considering that the environmentally relevant concentrations of 6-PPDQ were ≤10 µg/L, our data suggested that long-term exposure to 6-PPDQ at environmentally relevant concentrations potentially results in intestinal toxicity by disrupting functional state of intestinal barrier in organisms.

Influence of water quality factors on cake layer 3D structures and water channels during ultrafiltration process.

The three-dimensional (3D) structure of the cake layer, which could be influenced by water quality factors, plays a significant role in the ultrafiltration (UF) efficiency of water purification. However, it remains challenging to precisely reveal the variation of cake layer 3D structures and water channel characteristics. Herein, we systematically report the variation in the cake layer 3D structure at the nanoscale induced by key water quality factors and reveal its influence on water transport, in particular the abundance of water channels within the cake layer. In comparison with pH and Na+, Ca2+ played more significant role in determining cake layer structures. The sandwich-like cake layer, which was induced by the asynchronous deposition of humic acids and sodium alginate (SA), shifted to an isotropic structure when Ca2+ was present due to the Ca2+ bridging. In comparison with the sandwich-like structure, the isotropic cake layer has higher fractions of free volume (voids) and more water channels, leading to a 147% improvement in the water transport coefficient, 60% reduction in the cake layer resistance, and 21% increase in the final membrane specific flux. Our work elucidates a structure-property relationship where improving the isotropy of the cake layer 3D structure is conducive to the optimization of water channels and water transport within cake layers. This could inspire tailored regulation strategies for cake layers to enhance the UF efficiency of water purification.

An artificial bone filling material of poly l-lactic acid/collagen/nano-hydroxyapatite microspheres: Preparation and collagen regulation on the property.

Artificial bone materials are in great need due to a lot of bone injuries. Herein, collagen/nano-hydroxyapatite (Col/nHA, C-H) composite nanospheres were obtained by in-situ mineralization, and poly L-lactic acid/collagen/nano-hydroxyapatite (PLLA/Col/nHA, P-C-H) was further prepared by high-speed shear emulsification method. The interfacial properties and structure between PLLA and nHA are regulated by the adhesive property of Col. The morphology, structure and properties of P-C-H microsphere were characterized in detail by scanning electron microscopy (SEM), transmission electron microscopy (TEM), Brunauer-Emmett-Teller (BET) and simulated degradation of PBS in vitro. The results show that C-H is uniformly distributed in P-C-H microspheres, and a mesoporous material with a "pomegranate" structure and a particle size of 5-30 µm is self-assembled based on C-H multiple composite microspheres. It is beneficial to the sustained-release degradation of P-C-H and the retention/release of Ca2+. The 60-day PBS degradation shows that PLLA delays the degradation of nHA, making the degradation rate of P-C-H basically consist with the human bone healing cycle. The co-culture of P-C-H with MC3T3-E1 cells shows that P-C-H has high biocompatibility and no cytotoxicity. The cell viability is higher than 100 % in 72 h, indicating P-C-H has a proliferation effect on cell growth. Alkaline phosphatase and quantitative real-time PCR test show a positive promotion of P-C-H in cell proliferation and differentiation. The multi-layered P-C-H microspheres have an application potential in bone tissue engineering.

Cake layer 3D structure regulation to optimize water channels during Al-based coagulation-ultrafiltration process.

The variation in cake layer three-dimensional (3D) structures and related water channel characteristics induced by coagulation pretreatment remains unclear; however, gaining such knowledge will aid in improving ultrafiltration (UF) efficiency for water purification. Herein, the regulation of cake layer 3D structures (3D distribution of organic foulants within cake layers) by Al-based coagulation pretreatment was analyzed at the micro/nanoscale. The sandwich-like cake layer of humic acids and sodium alginate induced without coagulation was ruptured, and foulants were gradually uniformly distributed within the floc layer (toward an isotropic structure) with increasing coagulant dosage (a critical dosage was observed). Furthermore, the structure of the foulant-floc layer was more isotropic when coagulants with high Al13 concentrations were used (either AlCl3 at pH 6 or polyaluminum chloride, in comparison with AlCl3 at pH 8 where small-molecular-weight humic acids were enriched near the membrane). These high Al13 concentrations lead to a 48.4% higher specific membrane flux than that seen for UF without coagulation. Molecular dynamics simulations revealed that with increasing Al13 concentration (Al13: 6.2% to 22.6%), the water channels within the cake layer were enlarged and more connected, and the water transport coefficient was improved by up to 54.1%, indicating faster water transport. These findings demonstrate that facilitating an isotropic foulant-floc layer with highly connected water channels by coagulation pretreatment with high-Al13-concentration coagulants (having a strong ability to complex organic foulants) is the key issue in optimizing the UF efficiency for water purification. The results should provide further understanding of the underlying mechanisms of coagulation-enhancing UF behavior and inspire precise design of coagulation pretreatment to achieve efficient UF.

Role of HIV membrane in neutralization by two broadly neutralizing antibodies.

Induction of effective antibody responses against HIV-1 infection remains an elusive goal for vaccine development. Progress may require in-depth understanding of the molecular mechanisms of neutralization by monoclonal antibodies. We have analyzed the molecular actions of two rare, broadly neutralizing, human monoclonal antibodies, 4E10 and 2F5, which target the transiently exposed epitopes in the membrane proximal external region (MPER) of HIV-1 gp41 envelope during viral entry. Both have long CDR H3 loops with a hydrophobic surface facing away from the peptide epitope. We find that the hydrophobic residues of 4E10 mediate a reversible attachment to the viral membrane and that they are essential for neutralization, but not for interaction with gp41. We propose that these antibodies associate with the viral membrane in a required first step and are thereby poised to capture the transient gp41 fusion intermediate. These results bear directly on strategies for rational design of HIV-1 envelope immunogens.

Alteration in Wnt signaling mediates induction of transgenerational toxicity of polystyrene nanoplastics in C. elegans.

Polystyrene nanoparticles (PS-NPs) have a potential toxicity on offspring after the exposure. However, the molecular basis for PS-NP in inducing transgenerational toxicity remains largely unknown. In this study, the role and the underlying mechanism of germline Wnt signaling in regulating transgenerational toxicity of PS-NPs were determined using an in vivo animal model of Caenorhabditis elegans. Exposure to PS-NP (1-100 µg/L) increased expression of Wnt ligand LIN-44 and decreased expression of Wnt receptor MIG-1. After the exposure, the transgenerational PS-NP toxicity on locomotion behavior and brood size were inhibited in lin-44(RNAi) nematodes, while enhanced in mig-1(RNAi) nematodes. The resistance to transgenerational PS-NP toxicity induced by RNAi of lin-44 in P0 generation (P0-G) was inhibited by RNAi of mig-1 in F1-G. In addition, after PS-NP exposure, germline RNAi of lin-44 at P0-G could increase the mig-1 expression in F1-G. Exposure to PS-NP (1-100 µg/L) further decreased expressions of Dishevelled proteins of DSH-1/2, increased APC complex component APR-1, and decreased expression of BAR-1/ß-catenin. Meanwhile, transgenerational PS-NP toxicity was enhanced by RNAi of dsh-1, dsh-2, or bar-1 and inhibited by RNAi of apr-1, suggesting that the DSH-1/2-APR-1-BAR-1 signaling cascade acted downstream of Wnt receptor MIG-1 to control transgenerational PS-NP toxicity. Moreover, BAR-1 acted upstream of DVE-1 to activate mitochondrial unfolded protein response (mt UPR) against the transgenerational PS-NP toxicity. Our data highlights the potential link between alteration in germline Wnt signaling and induction of transgenerational nanoplastic toxicity in organisms.