Feedback regulation of ubiquitination and phase separation of HECT E3 ligases.

Lipid homeostasis is essential for normal cellular functions and dysregulation of lipid metabolism is highly correlated with human diseases including neurodegenerative diseases. In the ubiquitin-dependent autophagic degradation pathway, Troyer syndrome-related protein Spartin activates and recruits HECT-type E3 Itch to lipid droplets (LDs) to regulate their turnover. In this study, we find that Spartin promotes the formation of Itch condensates independent of LDs. Spartin activates Itch through its multiple PPAY-motif platform generated by self-oligomerization, which targets the WW12 domains of Itch and releases the autoinhibition of the ligase. Spartin-induced activation and subsequent autoubiquitination of Itch lead to liquid-liquid phase separation (LLPS) of the poly-, but not oligo-, ubiquitinated Itch together with Spartin and E2 both in vitro and in living cells. LLPS-mediated condensation of the reaction components further accelerates the generation of polyubiquitin chains, thus forming a positive feedback loop. Such Itch-Spartin condensates actively promote the autophagy-dependent turnover of LDs. Moreover, we show that the catalytic HECT domain of Itch is sufficient to interact and phase separate with poly-, but not oligo-ubiquitin chains. HECT domains from other HECT E3 ligases also exhibit LLPS-mediated the promotion of ligase activity. Therefore, LLPS and ubiquitination are mutually interdependent and LLPS promotes the ligase activity of the HECT family E3 ligases.

FBXO38 mediates PD-1 ubiquitination and regulates anti-tumour immunity of T cells.

Dysfunctional T cells in the tumour microenvironment have abnormally high expression of PD-1 and antibody inhibitors against PD-1 or its ligand (PD-L1) have become commonly used drugs to treat various types of cancer1-4. The clinical success of these inhibitors highlights the need to study the mechanisms by which PD-1 is regulated. Here we report a mechanism of PD-1 degradation and the importance of this mechanism in anti-tumour immunity in preclinical models. We show that surface PD-1 undergoes internalization, subsequent ubiquitination and proteasome degradation in activated T cells. FBXO38 is an E3 ligase of PD-1 that mediates Lys48-linked poly-ubiquitination and subsequent proteasome degradation. Conditional knockout of Fbxo38 in T cells did not affect T cell receptor and CD28 signalling, but led to faster tumour progression in mice owing to higher levels of PD-1 in tumour-infiltrating T cells. Anti-PD-1 therapy normalized the effect of FBXO38 deficiency on tumour growth in mice, which suggests that PD-1 is the primary target of FBXO38 in T cells. In human tumour tissues and a mouse cancer model, transcriptional levels of FBXO38 and Fbxo38, respectively, were downregulated in tumour-infiltrating T cells. However, IL-2 therapy rescued Fbxo38 transcription and therefore downregulated PD-1 levels in PD-1+ T cells in mice. These data indicate that FBXO38 regulates PD-1 expression and highlight an alternative method to block the PD-1 pathway.

Carbon availability and microbial activity manipulate the temperature sensitivity of anaerobic degradation in a paddy soil profile.

Soil contains a substantial amount of organic carbon, and its feedback to global warming has garnered widespread attention due to its potential to modulate atmospheric carbon (C) storage. Temperature sensitivity (Q10) has been widely utilized as a measure of the temperature-induced enhancement in soil organic carbon (SOC) decomposition. It is currently rare to incorporate Q10 of CO2 and CH4 into the study of waterlogged soil profiles and explore the possibility of artificially reducing Q10 in rice fields. To investigate the key drivers of Q10, we collected 0-1 m paddy soil profiles, and stratified the soil for submerged anaerobic incubation. The relationship between SOC availability, microbial activity, and the Q10 of CO2 and CH4 emissions was examined. Our findings indicate that as the soil layer deepens, soil C availability and microbial activity declined, and the Q10 of anaerobic degradation increased. Warming increased C availability and microbial activity, accompanied by weakened temperature sensitivity. The Q10 of CO2 correlated strongly with soil resistant C components, while the Q10 of CH4 was significantly influenced by labile substrates. The temperature sensitivity of CH4 (Q10 = 3.99) was higher than CO2 emissions (Q10 = 1.78), indicating the need for greater attention of CH4 in predicting warming's impact on anaerobic degradation in rice fields. Comprehensively assessing CO2 and CH4 emissions, the 20-40 cm subsurface soil is the most temperature-sensitive. Despite being a high-risk area for C loss and CH4 emissions, management of this soil layer in agriculture has the potential to reduce the threat of global warming. This study underscores the importance of subsurface soil in paddy fields, advocating greater attention in scientific simulations and predictions of climate change.

Twelve-year conversion of rice paddy to wetland does not alter SOC content but decreases C decomposition and N mineralization in Japan.

Land-use change worldwide has been driven by anthropogenic activities, which profoundly regulates terrestrial C and N cycles. However, it remains unclear how the dynamics and decomposition of soil organic C (SOC) and N respond to long-term conversion of rice paddy to wetland. Here, soil samples from five soil depths (0-25 cm, 5 cm/depth) were collected from a continuous rice paddy and an adjacent wetland (a rice paddy abandoned for 12 years) on Shonai Plain in northeastern Japan. A four-week anaerobic incubation experiment was conducted to investigate soil C decomposition and N mineralization. Our results showed that SOC in the wetland and rice paddy decreased with soil depth, from 31.02 to 19.66 g kg-1 and from 30.26 to 18.86 g kg-1, respectively. There was no significant difference in SOC content between wetland and rice paddy at any depth. Soil total nitrogen (TN) content in the wetland (2.61-1.49 g kg-1) and rice paddy (2.91-1.78 g kg-1) showed decreasing trend with depth; TN was significantly greater in the rice paddy than in the wetland at all depths except 20-25 cm. Paddy soil had significantly lower C/N ratios but significantly larger decomposed C (Dec-C, CO2 and CH4 production) and mineralized N (Min-N, net NH4+-N production) than wetland soil across all depths. Moreover, the Dec-C/Min-N ratio was significantly larger in wetland than in rice paddy across all depths. Rice paddy had higher exponential correlation between Dec-C and SOC, Min-N and TN than wetland. Although SOC did not change, TN decreased by 14.1% after the land-use conversion. The Dec-C and Min-N were decreased by 32.7% and 42.2%, respectively, after the12-year abandonment of rice paddy. Conclusively, long-term conversion of rice paddy to wetland did not distinctly alter SOC content but increased C/N ratio, and decreased C decomposition and N mineralization in 0-25 cm soil depth.

Liquid fertilizers produced by microwave-assisted acid hydrolysis of livestock and poultry wastes and their effects on hot pepper cultivation.

Liquid fertilizers (LFs) produced by microwave-assisted acid hydrolysis of livestock and poultry wastes were applied to potted hot pepper (Capsicum annuum L.) to evaluate their potential to be used as amino acid LFs. A preliminary experiment was conducted to determine the optimum acid-hydrolysis conditions for producing LFs from a mixture of pig hair and faeces (P) and another mixture of chicken feathers and faeces (C). Two LFs were produced under the optimum acid-hydrolysis conditions (acidification by sulphuric acid (7.5 mol L-1) in a microwave (200 W) for 90 minutes), and a commercial amino acid LF (Guo Guang (GG)) was used for comparison. P, C and GG fertilizers were tested in potted hot pepper cultivation at two doses, whereas no fertilizer application served as the control (CK). P and C fertilizers significantly increased the fruit yield compared with GG fertilizer, particularly at the higher dose. Moreover, the treatments improved the fruit vitamin C and soluble sugar contents in the order of C > P > GG compared with CK. These results could be attributed to more types of amino acids in C fertilizer than in P and GG fertilizers. The results also indicated that the prepared fertilizers could significantly increase the shoot and root dry weight, soil available nitrogen and phosphorus contents and nitrogen, phosphorus, and potassium (NPK) uptake by plants compared with CK. In conclusion, microwave-assisted acid hydrolysis could effectively convert unusable wastes into valuable fertilizers comparable or even superior to commercial fertilizers.

RNF217 regulates iron homeostasis through its E3 ubiquitin ligase activity by modulating ferroportin degradation.

Ferroportin (FPN), the body's sole iron exporter, is essential for maintaining systemic iron homeostasis. In response to either increased iron or inflammation, hepatocyte-secreted hepcidin binds to FPN, inducing its internalization and subsequent degradation. However, the E3 ubiquitin ligase that underlies FPN degradation has not been identified. Here, we report the identification and characterization of a novel mechanism involving the RNF217-mediated degradation of FPN. A combination of 2 different E3 screens revealed that the Rnf217 gene is a target of Tet1, mediating the ubiquitination and subsequent degradation of FPN. Interestingly, loss of Tet1 expression causes an accumulation of FPN and an impaired response to iron overload, manifested by increased iron accumulation in the liver together with decreased iron in the spleen and duodenum. Moreover, we found that the degradation and ubiquitination of FPN could be attenuated by mutating RNF217. Finally, using 2 conditional knockout mouse lines, we found that knocking out Rnf217 in macrophages increases splenic iron export by stabilizing FPN, whereas knocking out Rnf217 in intestinal cells appears to increase iron absorption. These findings suggest that the Tet1-RNF217-FPN axis regulates iron homeostasis, revealing new therapeutic targets for FPN-related diseases.

Injectable, Self-Healing and Multiple Responsive Histamine Modified Hyaluronic Acid Hydrogels with Potentialities in Drug Delivery, Antibacterial and Tissue Engineering.

Hydrogels are 3D network structures composed of physically or chemically crosslinked, hydrophilic molecules. Compared with conventional hydrogels with static and permanent network structures, injectable and responsive hydrogels generated from dynamic networks, have attracted increasing attention from various disciplines due to their wide-ranging applications in tissue engineering, drug delivery, soft robotics, etc. Herein, an injectable self-healing and multiple-responsive hyaluronic acid (HA)- histamine (His)/metal hydrogel is developed by modifying His onto HA and the subsequent, dynamic coordination between imidazole and metal ions. The pH-responsive and mechanical behaviors exhibited by the HA-His/metal hydrogels are tunable with the kinds and the concentrations of metal ions. The HA-His/Zr4+ hydrogels demonstrate a moldable capability at a neutral pH and a multi-stimulus-responsive capability when exposed to a weak alkaline environment and hyaluronidase, which inhibits bacterial growth and biofilm formation. Biocompatibilities and accelerated wound healing are demonstrated in vitro and in vivo and are thoroughly investigated and well characterized. The HA-His/Zr4+ hydrogel has great potential in various biomedical applications, such as pH- and hyaluronidase-responsive sustained release, antibacterial, and implantable materials for tissue engineering.

Soil pH management for mitigating N2O emissions through nosZ (Clade I and II) gene abundance in rice paddy system.

Soil nitrous oxide (N2O) is produced by abiotic and biotic processes, but it is solely consumed by denitrifying microbes-encoded by nosZ genes. The nosZ gene includes two groups i.e. Clade I and Clade II, which are highly sensitive to pH. Managing pH of acidic soils can substantially influence soil N2O production or consumption through nosZ gene abundance. Nevertheless, the response of nosZ (Clade I and Clade II) to pH management needs elucidation in acidic soils. To clarify this research question, a pot experiment growing rice crop was conducted with three treatments: control (only soil), low dose of dolomite (LDD), and high dose of dolomite (HDD). The soil pH increased from 5.41 to 6.23 in the control, 6.5 in LDD and 6.8 in HDD treatment under flooded condition. The NH4+ and NO3- contents increased and reached the maximum at 30.4 and 21.5 mg kg-1, respectively, in HDD treatment under flooding condition. The contents of dissolved organic carbon and microbial biomass carbon showed a swift rise at midseason aeration and reached maximum at 30.7 and 101 mg kg-1 in the HDD treatment. Clade I, Clade II and 16S rRNA genes abundance increased with the onset of flooding, and occurred maximum in the HDD treatment. A peak in N2O emissions (5.96 µg kg-1 h-1) occurred at midseason events in the control when no dolomite was added. Dolomite application significantly (p ≤ 0.001) suppressed N2O emissions, and HDD treatment was more effective in reducing emissions. Pearson correlation, linear regressions and principal component analysis displayed that increased soil pH and Clade I and Clade II were the main controlling factors for N2O emission mitigation in acidic soil. This research demonstrates that ameliorating soil acidity with dolomite application is a potential option for the mitigation of N2O emissions.

Ascription of nosZ gene, pH and copper for mitigating N2O emissions in acidic soils.

Soil nitrous oxide (N2O) emissions are alarming for global warming and climate change. N2O reduction is carried out only by nosZ gene encoded N2O-reductase, which is highly sensitive to acidic pH and copper (Cu) contents. Therefore, a microcosm study was conducted to examine the attribution of soil pH management, Cu supply and nosZ gene abundance for N2O emission mitigation. Cu was applied at the dose of 0, 10, 25 and 50 mg kg-1 to three acidic soils (Soil 1, 2 and 3) without and with dolomite (0 and 5 g kg-1). Cu application and soil pH increment substantially enlarged the abundance of nosZ gene, and consequently mitigated soil N2O emissions; highest reduction with 25 Cu mg kg-1. Decline in NH4+ and subsequently accumulation of NO3-, and large contents of MBC and DOC in dolomite treated soils led to a substantial N2O reduction. The cumulative N2O emissions were lowest in the treatment of 25 Cu mg kg-1 with dolomite application for each soil. Results suggest that soil pH increment, an adequate Cu supply, and nosZ gene abundance can potentially lower soil N2O emissions in acidic soils.

Enzyme activities and organic matter mineralization in response to application of gypsum, manure and rice straw in saline and sodic soils.

Saline and alkaline soils are a challenge for sustainable crop production. The use of organic and inorganic amendments is a common practice to increase the fertility of salt-affected soils that can trigger faster carbon (C) and nitrogen (N) cycling. We examined the effects of gypsum (Gyps), farm manure (Manure) and rice straw (Straw) on enzyme activities, organic matter mineralization and CO2 emissions in two salt-affected soils [Solonchak (saline); pH: 8, electrical conductivity (EC): 6.5, sodium adsorption ratio (SAR): 2.5, and Solonetz (alkaline sodic); pH: 8.9, EC: 1.6, SAR: 17]. Gypsum addition decreased soil pH up to 0.62 and 0.30 units, SAR 1.2 and 5.2 units, and EC 2.9 and 1.4 units in Solonchak and Solonetz, respectively. Dissolved organic C, microbial biomass C, dissolved organic N, mineral N (NO3- and NH4+), enzyme activities (urease, invertase, catalase, phosphatase, phenol-oxidase), alkali extractable phenols, and available phosphorous increased with the application of all amendments in both soils. Solonetz released more CO2 than Solonchak, whereas maximum CO2 emissions were common after manure application (3140 mg kg-1 in Solonchak, and 3890 mg kg-1 in Solonetz). We conclude that high SAR and low EC increase CO2 emissions through accelerated C and N cycling and manure decomposition in Solonetz soils.

MKRN3-mediated ubiquitination of Poly(A)-binding proteins modulates the stability and translation of GNRH1 mRNA in mammalian puberty.

The family of Poly(A)-binding proteins (PABPs) regulates the stability and translation of messenger RNAs (mRNAs). Here we reported that the three members of PABPs, including PABPC1, PABPC3 and PABPC4, were identified as novel substrates for MKRN3, whose deletion or loss-of-function mutations were genetically associated with human central precocious puberty (CPP). MKRN3-mediated ubiquitination was found to attenuate the binding of PABPs to the poly(A) tails of mRNA, which led to shortened poly(A) tail-length of GNRH1 mRNA and compromised the formation of translation initiation complex (TIC). Recently, we have shown that MKRN3 epigenetically regulates the transcription of GNRH1 through conjugating poly-Ub chains onto methyl-DNA bind protein 3 (MBD3). Therefore, MKRN3-mediated ubiquitin signalling could control both transcriptional and post-transcriptional switches of mammalian puberty initiation. While identifying MKRN3 as a novel tissue-specific translational regulator, our work also provided new mechanistic insights into the etiology of MKRN3 dysfunction-associated human CPP.

Fe(III) stabilizing soil organic matter and reducing methane emissions in paddy fields under varying flooding conditions.

The role of iron (Fe) in soil organic matter (SOM) stabilization and decomposition in paddy soils has recently gained attention, but the underlying mechanisms during flooding and drying periods remain elusive. As the depth water layer is maintained in the fallow season, there will be more soluble Fe than during the wet and drainage seasons and the availability of oxygen (O2) will be different. To assess the influence of soluble Fe on SOM mineralization during flooding, an incubation experiment was designed under oxic and anoxic flooding conditions, with and without Fe(III) addition. The results showed that Fe(III) addition significantly (p < 0.05) decreased SOM mineralization by 14.4 % under oxic flooding conditions over 16 days. Under anoxic flooding incubation, Fe(III) addition significantly (p < 0.05) decreased 10.8 % SOM decomposition, mainly by 43.6 % methane (CH4) emission, while no difference in carbon dioxide (CO2) emission was noticed. These findings suggest that implementing appropriate water management strategies in paddy soils, considering the roles of Fe under both oxic and anoxic flooding conditions, can contribute to SOM preservation and mitigation of CH4 emissions.

Impact of bentonite on greenhouse gas emissions during pig manure composting and its subsequent application.

Additives were widely investigated to retain the nutrients and mitigate the greenhouse gas emissions (GHGs) during manure composting. However, the sustained effects of additives on the GHGs emissions following incorporation of composts to soil were scarcely explored. This study evaluated the effects of bentonite added at the beginning of pig manure composting on the GHGs emissions during two successive processes, i.e., composting and soil incubation amended with composting products. Addition of bentonite did not hinder the composting process and alter the total CO2 emission. On the other hand, reduction by about 17% and 29% for CH4 and N2O emission, respectively, was achieved in the presence of bentonite during composting. Incorporation of the final composting products to soil enhanced significantly the soil C and N of various forms, and gas emissions of CO2 and N2O. However, no significant differences were observed between bentonite-manure co-compost and manure-only compost application except for the N2O emission. Compared to the manure-only compost, compost amended with bentonite reduced N2O loss by around 6.8%, but not statistically significant. This study confirmed that addition of bentonite at the composting stage can mitigate the GHGs emission considering both composting and compost application stages, with all reductions occurring at the composting stage.

A combination of organic fertilizers partially substitution with alternate wet and dry irrigation could further reduce greenhouse gases emission in rice field.

Alternate wet and dry (AWD) irrigation and organic fertilizers substitution (OFS) have contrasting effects on CH4 and N2O emissions in rice cultivation. Combining these two practices may be feasible for simultaneous reduction of CH4 and N2O emission from paddy. Hence, we conducted a two-year field experiment to explore the reduction of greenhouse gases under the combination of AWD and OFS. The field experiment which was designed with two irrigation methods (continuous flooding (CF) irrigation and AWD irrigation), and five nitrogen regimes (N0, N135, and N180 represent 0, 135, and 180 kg N ha-1, respectively, ON25 and ON50 represent 25% and 50% OFS for inorganic fertilizer, respectively). The results showed a single-peak emission for CH4 fluxes during the whole rice growing season in all water and nitrogen treatments while the N2O fluxes peak only observed during tillering period with AWD irrigation. AWD irrigation and OFS showed a limited reduction in global warming potential (GWP). These were owing to that AWD irrigation reduced 38.3% CH4 emissions while increase 145.9% N2O emissions when compared to CF irrigation, and the low rate (25%) OFS only reduced CH4 emission by 29.4% while high rate (50%) only reduce N2O emission by 38.8% when compared to conventional inorganic nitrogen fertilizer (N180). Combined AWD and ON25 could maximize the reduction in GWP and yield-scaled GWP, which were reduce 58.0% and 52.5%, respectively, compare to the conventional water and nitrogen management (CF and N180). Furthermore, the structural equation modelling (SEM) indicated that the soil dissolved organic carbon (DOC) and rice aboveground biomass showed a significant positive effect on CH4 fluxes while soil NH4+ with a negative effect, and the soil NH4+, nitrification potential, denitrification potential significant affected N2O fluxes with a positive effect while DOC with a negative effect. These results investigated that 25% OFS rate for inorganic fertilizer could further reduce warming potential in AWD irrigation rice field.

Opposing roles of E3 ligases TRIM23 and TRIM21 in regulation of ion channel ANO1 protein levels.

Anoctamin-1 (ANO1) (TMEM16A) is a calcium-activated chloride channel that plays critical roles in diverse physiological processes, such as sensory transduction and epithelial secretion. ANO1 levels have been shown to be altered under physiological and pathological conditions, although the molecular mechanisms that control ANO1 protein levels remain unclear. The ubiquitin-proteasome system is known to regulate the levels of numerous ion channels, but little information is available regarding whether and how ubiquitination regulates levels of ANO1. Here, we showed that two E3 ligases, TRIM23 and TRIM21, physically interact with the C terminus of ANO1. In vitro and in vivo assays demonstrated that whereas TRIM23 ubiquitinated ANO1 leading to its stabilization, TRIM21 ubiquitinated ANO1 and induced its degradation. Notably, ANO1 regulation by TRIM23 and TRIM21 is involved in chemical-induced pain sensation, salivary secretion, and heart-rate control in mice, and TRIM23 also mediates ANO1 upregulation induced by epidermal growth factor treatment. Our results suggest that these two antagonistic E3 ligases act together to control ANO1 expression and function. Our findings reveal a previously unrecognized mechanism for regulating ANO1 protein levels and identify a potential molecular link between ANO1 regulation, epidermal growth factor, and other signaling pathways.

Effects of water regimes on soil N2O, CH4 and CO2 emissions following addition of dicyandiamide and N fertilizer.

Water regimes strongly impact soil C and N cycling and the associated greenhouse gases (GHGs, i.e., CO2, CH4 and N2O). Therefore, a study was conducted to examine the impacts of flooding-drying of soil along with application of nitrogen (N) fertilizer and nitrification inhibitor dicyandiamide (DCD) on GHGs emissions. This study comprised four experimental treatments, including (i) control (CK), (ii) dicyandiamide, 20 mg kg-1 (DCD), (iii) nitrogen fertilizer, 300 mg kg-1 (N) and (iv) DCD + N. All experimental treatments were kept under flooded condition at the onset of the experiment, and then converted to 60% water filled pore space (WFPS). At flooding stage, N2O emissions were lower as compared to 60% WFPS. The highest cumulative N2O emission was 0.98 mg N2O-N kg-1 in N treated soil due to high substrates of mineral N contents, but lowest (0.009 mg N2O-N kg-1) in the DCD treatment. The highest cumulative CH4 emissions (80.54 mg CH4-C kg-1) were observed in the N treatment, while uptake of CH4 was observed in the DCD treatment. As flooded condition converted to 60% WFPS, CO2 emissions gradually increased in all experimental treatments, but the maximum cumulative CO2 emission was 477.44 mg kg-1 in the DCD + N treatment. The maximum dissolved organic carbon (DOC) contents were observed in N and DCD + N treatments with the values of 57.12 and 58.92 mg kg-1, respectively. Microbial biomass carbon (MBC) contents were higher at flooding while lower at transition phase, and increased at the initiation of 60% WFPS stage. However, MBC contents declined at the later stage of 60% WFPS. The maximum MBC contents were 202.12 and 192.41 mg kg-1 in N and DCD + N treatments, respectively. Results demonstrated that water regimes exerted a dramatic impact on C and N dynamics, subsequently GHGs, which were highly controlled by DCD at both flooding and 60% WFPS conditions.

Earthworms promote the transfer of 15N-urea to lettuce while limit appreciably increase 15N losing to environment.

Earthworm activities not only increase nitrogen (N) uptake by crops, but also lead to N losing to environment. Thus, the present study examined the transformation of 15N-labeled urea with and without earthworms (Metaphire guillelmi) in a soil-lettuce system. We evaluated lettuce 15N uptake, 15N losses including N2O emission, NH3 volatilization and leaching, as well as 15N remaining in soil. Results showed that 15N-urea uptakes by lettuce significantly increased from 33.07% to 42.72% with earthworm presence. However, little difference was found on the total amounts of leaching and gaseous losses (N2O emission and NH3 volatilization) from 15N-urea between the treatment with and without earthworms (4.04 and 5.38%, respectively). Most of the 15N-urea remained in the soil, accounting for 48.44% and 60.65% of the 15N-urea in soil with and without earthworm presence. We conclude that earthworms enhanced the transfer of 15N-urea to lettuce without appreciably increasing the 15N-urea loss from soil to the environment.

Early life exposure to triclosan from antimicrobial daily necessities may increase the potential risk of autism spectrum disorder: A multicenter study in China.

BACKGROUND: Autism spectrum disorders (ASD) are a group of complex neurodevelopmental disorders with unclear etiologies. Our recent work indicated that maternal exposure to triclosan (TCS) significantly increased the autistic-like behavior in rats, possibly through disrupting neuronal retinoic acid signaling. Although environmental endocrine disruptors (EEDs) have been associated with autism in humans, the relationship between TCS, one of the EEDs found in antibacterial daily necessities, and autism has received little attention. OBJECTIVE: The aims of this multicenter study were to evaluate TCS concentrations in typically developing (TD) children and ASD children, and to determine the relationship between TCS levels and the core symptoms of ASD children. METHODS: A total of 1345 children with ASD and 1183 TD children were enrolled from 13 cities in China. Ages ranged between 2 and 7 years. A questionnaire was used to investigate the maternal use of antibacterial daily necessities (UADN) during pregnancy. The core symptoms of ASD were evaluated using the Autism Behavior Checklist (ABC), Childhood Autism Rating Scale (CARS), Social Response Scale (SRS), and the Children Neuropsychological and Behavior Scale-Revision 2016 (CNBS-R2016). The TCS concentration was measured using LC-MS/MS. RESULTS: Maternal UADN during pregnancy may be an unrecognized potential environmental risk factor for ASD (OR=1.267, P = 0.023). Maternal UADN during pregnancy strongly correlated with TCS levels in the offspring (Adjusted ß = 0.277, P < 0.001). TCS concentration was higher in ASD children (P = 0.005), and positively correlated with ABC (Sensory subscales: P = 0.03; Social self-help subscales: P = 0.011) and SRS scale scores (Social awareness subscales: P = 0.045; Social communication subscales: P = 0.001; Autism behavior mannerisms subscales: P = 0.006; SRS total score: P = 0.003) in ASD children. This association was more pronounced in boys than in girls. CONCLUSION: To our knowledge, this is the first case-control study to examine the correlation between TCS and ASD. Our results suggest that maternal UADN during pregnancy may be a potential risk of ASD in offspring. Further detection of TCS levels showed that maternal UADN during pregnancy may be associated with excessive TCS exposure. In addition, the level of TCS in children with ASD is higher than TD children. The higher levels of TCS in children with ASD may be significantly associated with more pronounced core symptoms, and this association was more significant in male children with ASD.

Translatomic profiling reveals novel self-restricting virus-host interactions during HBV infection.

BACKGROUND & AIMS: HBV remains a global threat to human health. It remains incompletely understood how HBV self-restricts in the host during most adult infections. Thus, we performed multi-omics analyses to systematically interrogate HBV-host interactions and the life cycle of HBV. METHODS: RNA-sequencing and ribosome profiling were conducted with cell-based models for HBV replication and gene expression. The novel translational events or products hereby detected were then characterized, and functionally assessed in both cell and mouse models. Moreover, quasi-species analyses of HBV subpopulations were conducted with patients at immune tolerance or activation phases, using next- or third-generation sequencing. RESULTS: We identified EnhI-SL (Enhancer I-stem loop) as a new cis element in the HBV genome; mutations disrupting EnhI-SL were found to elevate viral polymerase expression. Furthermore, while re-discovering HpZ/P', a previously under-explored isoform of HBV polymerase, we also identified HBxZ, a novel short isoform of HBX. Having confirmed their existence, we functionally characterized them as potent suppressors of HBV gene expression and genome replication. Mechanistically, HpZ/P' was found to repress HBV gene expression partially by interacting with, and sequestering SUPV3L1. Activation of the host immune system seemed to reduce the abundance of HBV mutants deficient in HpZ/P' or with disruptions in EnhI-SL. Finally, SRSF2, a host RNA spliceosome protein that is downregulated by HBV, was found to promote the splicing of viral pre-genomic RNA and HpZ/P' biogenesis. CONCLUSION: This study has identified multiple self-restricting HBV-host interactions. In particular, SRSF2-HpZ/P' appeared to constitute another negative feedback mechanism in the HBV life cycle. Targeting host splicing machinery might thus represent a strategy to intervene in HBV-host interactions. LAY SUMMARY: There remain many unknowns about the natural history of HBV infection in adults. Herein, we identified new HBV-host mechanisms which could be responsible for self-restricting infections. Targeting these mechanisms could be a promising strategy for the treatment of HBV infections.

Regulation of O-Linked N-Acetyl Glucosamine Transferase (OGT) through E6 Stimulation of the Ubiquitin Ligase Activity of E6AP.

Glycosyltransferase OGT catalyzes the conjugation of O-linked ß-D-N-acetylglucosamine (O-GlcNAc) to Ser and Thr residues of the cellular proteins and regulates many key processes in the cell. Here, we report the identification of OGT as a ubiquitination target of HECT-type E3 ubiquitin (UB) ligase E6AP, whose overexpression in HEK293 cells would induce the degradation of OGT. We also found that the expression of E6AP in HeLa cells with the endogenous expression of the E6 protein of the human papillomavirus (HPV) would accelerate OGT degradation by the proteasome and suppress O-GlcNAc modification of OGT substrates in the cell. Overall, our study establishes a new mechanism of OGT regulation by the ubiquitin-proteasome system (UPS) that mediates the crosstalk between protein ubiquitination and O-GlcNAcylation pathways underlying diverse cellular processes.