Construction of amine-rich coating utilizing amine curing reaction of triglycidyl isocyanate with triethylenetetramine on the surface of poly(styrene-divinylbenzene) microspheres for preparation of anion exchange stationary phase.

Epoxy resins, as important thermosetting polymers, exhibit excellent adhesion to various substrates. In view of this, reticulate coating of triglycidyl isocyanate with triethylenetetramine was introduced onto the surface of poly(styrene-divinylbenzene) utilizing amine curing reaction to obtain poly(styrene-divinylbenzene)@triglycidyl isocyanate-triethylenetetramine composite microspheres. The amino groups and epoxy groups of triglycidyl isocyanate-triethylenetetramine endowed poly(styrene-divinylbenzene) with good reactivity, which could be quaternized under mild conditions to obtain an anion exchange chromatographic stationary phase. The quaternized poly(styrene-divinylbenzene)@triglycidyl isocyanate-triethylenetetramine was characterized by scanning electron microscope, Fourier-transform infrared spectroscopy, N2 adsorption-desorption experiment, et al. The chromatographic performance of the customized column was evaluated by separating seven conventional anions, organic weak acids, and carbohydrates. Poly(styrene-divinylbenzene)@triglycidyl isocyanate-triethylenetetramine possesses the uniform size of poly(styrene-divinylbenzene) microspheres and good reactivity of triglycidyl isocyanate-triethylenetetramine, which offers a flexible strategy for the preparation of anion exchange stationary phase. The column exhibits excellent chemical and mechanical stability and chromatographic performance. Finally, the column was successfully applied for the determination of nitrite in pickles.

Transoral Robotic Surgery and Human Papillomavirus Infection: Impact on Oropharyngeal Cancer Prognosis.

Background/Objective: The incidence of oropharyngeal cancer (OPC) remains significant, with a rising prevalence of HPV-positive (HPV+) cases, underscoring the growing importance of appropriate treatment approaches for this condition. While HPV+ OPC typically exhibits a more favorable prognosis than HPV-negative (HPV-) OPC, certain HPV+ OPC patients still face adverse outcomes. This study aimed to assess the effectiveness of TORS versus traditional surgery in treating OPC patients and investigate the prognostic implications of specific variants in the HPV genome. Methods: The clinical information, including pathological features, treatments, and outcomes (death), of 135 OPC patients treated with traditional surgery from 2008 to 2018 (the non-TORS group) and 130 OPC patients treated with TORS from 2017 to 2021 (the TORS group) were obtained from Sun Yat-sen University Cancer Center (SYSUCC). A comparative analysis of 3-year overall survival (OS) was performed between these two groups. Furthermore, we conducted next-generation sequencing for the HPV16 genome of the 68 HPV+ OPC cases to characterize single-nucleotide variations (SNVs) in the HPV16 genome and evaluate its association with HPV+ OPC patient survival. Results: The comparative analysis of 3-year OS between the two groups (TORS vs. non-TORS) revealed a significant prognostic improvement in the TORS group for OPC patients with a T1-T2 classification (89.3% vs. 72.0%; p = 1.1 × 10-2), stages I-II (92.1% vs. 82.2%; p = 4.6 × 10-2), and stages III-IV (82.8% vs. 62.2%; p = 5.7 × 10-2) and for HPV- patients (85.5% vs. 33.3%; p < 1.0 × 10-6). Furthermore, three SNVs (SNV1339A>G, SNV1950A>C, and SNV4298A>G) in the HPV16 genome were identified as being associated with worse survival. These SNVs could alter protein interactions and weaken the binding affinity for MHC-II, promoting viral amplification and immune evasion. Conclusions: TORS exhibited a superior prognosis to traditional surgery in OPC patients. Additionally, identifying specific SNVs within the HPV16 genome provided potential prognostic markers for HPV+ OPC. These significant findings hold clinical relevance for treatment decision-making and prognostic assessment in patients with OPC.

Model-Based Sensitivity Analysis of the Temperature in Laser Powder Bed Fusion.

To quantitatively evaluate the effect of the process parameters and the material properties on the temperature in laser powder bed fusion (LPBF), this paper proposed a sensitivity analysis of the temperature based on the validated prediction model. First, three different heat source modes-point heat source, Gaussian surface heat source, and Gaussian body heat source-were introduced. Then, a case study of Ti6Al4V is conducted to determine the suitable range of heat source density for the three different heat source models. Based on this, the effects of laser processing parameters and material thermophysical parameters on the temperature field and molten pool size are quantitatively discussed based on the Gaussian surface heat source. The results indicate that the Gaussian surface heat source and the Gaussian body heat source offer higher prediction accuracy for molten pool width compared to the point heat source under similar processing parameters. When the laser energy density is between 40 and 70 J/mm3, the prediction accuracy of the Gaussian surface heat source and the body heat source is similar, and the average prediction errors are 4.427% and 2.613%, respectively. When the laser energy density is between 70 and 90 J/mm3, the prediction accuracy of the Gaussian body heat source is superior to that of the Gaussian surface heat source. Among the influencing factors, laser power exerts the greatest influence on the temperature field and molten pool size, followed by scanning speed. In particular, laser power and scan speed contribute 38.9% and 23.5% to the width of the molten pool, 39.1% and 19.6% to the depth of the molten pool, and 38.9% and 21.5% to the maximum temperature, respectively.

Effects of catechol grafting on chitosan-based coacervation and adhesion.

In this study, we investigated detailedly the contribution of catechol in tuning the formation and adhesive properties of coacervates. We have constructed a series of catechol-grafted Chitosan (Chitosan-C), and investigated their coacervation with gum arabic (GA) and the corresponding adhesion. We demonstrate that, increasing catechol grafting ratio from 0 %-44 % impacted the coacervation moderately, while enhanced the adhesion of the coacervate up to 438 % when the catechol faction was 37 %. Further increasing the grafting ratio to 55 % led to precipitated coacervates associated with a declined adhesion. Our findings identify the optimal grafting threshold for coacervation and adhesion, providing insights into the underlying mechanism of coacervate binding. Moreover, the catechol enhancement on adhesion of coacervates tolerates different substrates and diverse polyelectrolyte pairs. The revealed principles shall be helpful for designing adhesive coacervates and boosting their applications in various industrial and biomedical areas.

Laparoscopic treatment of colonic endometriosis causing periodic abdominal pain and hematochezia: A case report.

RATIONALE: Endometriosis, a benign disease, has a malignant biological behavior and is highly prone to recurrence. Although gastrointestinal involvement is the most common site for extra-genital endometriosis, deep infiltrative endometriosis, which affects the mucosal layer, is very rare. PATIENT CONCERNS: A 44-year-old woman with a 6-month history of recurring abdominal pain and Hematochezia. The patient visited several hospitals over the past six months and was suspected to have been diagnosed with a digestive disease, for which medication was ineffective, leading to a great deal of anxiety. DIAGNOSES: Colonic endometriosis. INTERVENTIONS: After a thorough imaging evaluation and preoperative discussion, laparoscopic colonic endometriosis resection under indocyanine green indication was performed by gynecologists and gastroenterologists. OUTCOMES: After laparoscopic treatment, the patient's symptoms improved significantly, with occasional pain felt and no blood in the stool. LESSONS: This case provides a rare example of sigmoid endometriosis causing periodic abdominal pain and Hematochezia. We report a clinical case to investigate the feasibility of an indocyanine green fluorescent contrast technique to guide the scope of surgery in laparoscopic deep infiltrative endometriosis surgery. In intestinal endometriosis surgery, indocyanine green fluoroscopy may indicate the lesion's precise localization.

ARF3-Mediated Regulation of SPL in Early Anther Morphogenesis: Maintaining Precise Spatial Distribution and Expression Level.

Early anther morphogenesis is a crucial process for male fertility in plants, governed by the transcription factor SPL. While the involvement of AGAMOUS (AG) in SPL activation and microsporogenesis initiation is well established, our understanding of the mechanisms governing the spatial distribution and precise expression of SPL during anther cell fate determination remains limited. Here, we present novel findings on the abnormal phenotypes of two previously unreported SPL mutants, spl-4 and spl-5, during anther morphogenesis. Through comprehensive analysis, we identified ARF3 as a key upstream regulator of SPL. Our cytological experiments demonstrated that ARF3 plays a critical role in restricting SPL expression specifically in microsporocytes. Moreover, we revealed that ARF3 directly binds to two specific auxin response elements on the SPL promoter, effectively suppressing AG-mediated activation of SPL. Notably, the arf3 loss-of-function mutant exhibits phenotypic similarities to the SPL overexpression mutant (spl-5), characterized by defective adaxial anther lobes. Transcriptomic analysis revealed differential expression of the genes involved in the morphogenesis pathway in both arf3 and spl mutants, with ARF3 and SPL exhibited opposing regulatory effects on this pathway. Taken together, our study unveils the precise role of ARF3 in restricting the spatial expression and preventing aberrant SPL levels during early anther morphogenesis, thereby ensuring the fidelity of the critical developmental process in plants.

PXL1 and SERKs act as receptor-coreceptor complexes for the CLE19 peptide to regulate pollen development.

Gametophyte development in angiosperms occurs within diploid sporophytic structures and requires coordinated development; e.g., development of the male gametophyte pollen depends on the surrounding sporophytic tissue, the tapetum. The mechanisms underlying this interaction remain poorly characterized. The peptide CLAVATA3/EMBRYO SURROUNDING REGION-RELATED 19 (CLE19) plays a "braking" role in preventing the harmful overexpression of tapetum transcriptional regulators to ensure normal pollen development in Arabidopsis. However, the CLE19 receptor is unknown. Here, we show that CLE19 interacts directly with the PXY-LIKE1 (PXL1) ectodomain and induces PXL1 phosphorylation. PXL1 is also required for the function of CLE19 in maintaining the tapetal transcriptional regulation of pollen exine genes. Additionally, CLE19 induces the interactions of PXL1 with SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE (SERK) coreceptors required for pollen development. We propose that PXL1 and SERKs act as receptor and coreceptor, respectively, of the extracellular CLE19 signal, thereby regulating tapetum gene expression and pollen development.

Expression profiles and potential roles of serum tRNA­derived fragments in diabetic nephropathy.

Diabetic nephropathy (DN) is one of the most important causes of end-stage renal disease and current treatments are ineffective in preventing its progression. Transfer RNA (tRNA)-derived fragments (tRFs), which are small non-coding fragments derived from tRNA precursors or mature tRNAs, have a critical role in various human diseases. The present study aimed to investigate the expression profile and potential functions of tRFs in DN. High-throughput sequencing technology was employed to detect the differential serum levels of tRFs between DN and diabetes mellitus and to validate the reliability of the sequencing results using reverse transcription-quantitative PCR. Ultimately, six differentially expressed (DE) tRFs were identified (P<0.05; |log2fold change| ≥1), including three upregulated (tRF5-GluCTC, tRF5-AlaCGC and tRF5-ValCAC) and three downregulated tRFs (tRF5-GlyCCC, tRF3-GlyGCC and tRF3-IleAAT). Potential functions and regulatory mechanisms of these DE tRFs were further evaluated using an applied bioinformatics-based analysis. Gene ontology analysis revealed that the DE tRFs are mainly enriched in biological processes, including axon guidance, Rad51 paralog (Rad51)B-Rad51C-Rad51D-X-Ray repair cross-complementing 2 complex, nuclear factor of activated T-cells protein binding and fibroblast growth factor-activated receptor activity. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis indicated that they are associated with axon guidance, neurotrophin signaling, mTOR signaling, AMPK signaling and epidermal growth factor receptor family signaling pathways. In conclusion, the present findings indicated that tRFs were DE in DN and may be involved in the regulation of DN pathology through multiple pathways, thereby providing a new perspective for the study of DN therapeutic targets.

Clinical diagnostic value of liquid chromatography-tandem mass spectrometry method for primary aldosteronism in patients with hypertension: A systematic review and meta-analysis.

Background: Primary aldosteronism (PA) is currently considered the most common cause of secondary and endocrine hypertension. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) as a new detection technique has been gradually applied in the diagnosis of PA. However, the diagnostic value of LC-MS/MS methods for PA has not been systematically clinically validated. The aim was to access the diagnostic accuracy, sensitivity, and specificity of LC-MS/MS methods as screening tools in PA. Materials and methods: A literature search of PubMed, Embase, Medline, Web of Science, Scopus, Science Direct, and Chinese databases was carried out to June 2022 with no language restriction. Data on sensitivity and specificity and other evaluation indicators were extracted and pooled with STATA and Meta-disc software. Heterogeneity was evaluated and meta-regression and subgroup analysis was performed to elucidate sources of heterogeneity. Results: 12 studies of the diagnostic test were suitable and included in the meta-analysis. Pooled sensitivity, specificity, and diagnostic odds ratio were 0.89 (95% CI: 0.83-0.93), 0.87 (95% CI: 0.82-0.91), and 55 (95% CI: 28-110), respectively. Subgroup analysis assessed the diagnostic power of LC-MS/MS based on the type of detection index. ARR and PAC based on LC-MS/MS methods have the higher diagnostic value compared with other indices, diagnostic odds ratios were 121.65 (95% CI: 36.28-407.98) and 49.85 (95% CI: 24.87-99.93). There was considerable heterogeneity among studies. Conclusion: LC-MS/MS methods had higher accuracy and reliability in the diagnosis of primary aldosteronism. LC-MS/MS-based ARR and PAC can be further promoted and applied in the diagnosis of primary aldosteronism.

Effect of grazing and nitrogen addition on the occurrence of species with different seed masses in alpine meadows on the Tibet Plateau.

Seed mass (SM) is a core functional trait of plant species. Thus, information of the effect of grazing and nitrogen addition on the occurrence of species with different SMs can help us understand the influence of grazing and fertilization on survival of species and community assembly. In alpine meadows with different grazing and nitrogen addition treatments on the eastern Tibet Plateau, we measured SM and plant height (H) of the most common plants and conducted a series of quadrat surveys. Overall, grazing promoted survival of small-seeded species, while fertilization suppressed their survival. At the community level, moderate grazing reduced the average community weighted mean (CWM) of SM and increased the coefficient of variation (CV) of SM. However, there was no significant difference between the functional diversity (FDrao, calculated as Rao's index) of SM in grazed and nongrazed meadows. Nitrogen addition significantly increased the CWM of SM, had a marginally significant effect on FDrao of SM, but had no significant effect on CV of SM. The impact of gazing and fertilization on occurrence of species with different SMs can be explained by their effect on vegetation height. From the perspective of SM selection, our study helps clarify the mechanism of species diversity loss due to fertilization, and that of species diversity increase by moderate grazing.

Regional gradients in intraspecific seed mass variation are associated with species biotic attributes and niche breadth.

Quantifying intraspecific trait variation (ITV) is crucial for understanding species local adaptation and regional distribution. Intraspecific seed mass variation (ITVsm) is expected to vary with environmental gradients or co-vary with related biotic attributes, but these relationships are not well known in a multispecies space. We performed interspecific and phylogenetic analyses to evaluate the relative power of three species biotic attributes and four niche breadth traits in explaining ITVsm variation for 434 eastern Qinghai-Tibetan species. We showed a positive relationship between species' ITVsm and their niche breadth in the light, moisture and disturbance dimensions, supporting the idea that high ITV allows species to match their traits to different habitat conditions and thus to distribute in a wide range of environments. However, we did find significant direct effect of species' thermal niche on individual seed mass variation. Meanwhile, we showed significant effects of seed dispersal mode, but not of life form and pollination type, on ITVsm. This suggests that the covariation or co-evolution between seed and disperser was related to the pattern and magnitude of ITVsm, but not to plant lifespan, the quality and allocation pattern of available resources and the availability of pollination vector. Lastly, all multivariate models showed a significant combined contribution of species' biotic attributes and niche breadth to their ITVsm, implying that intrinsic biotic limitations and extrinsic abiotic pressures may operate simultaneously in controlling regional-scale intraspecific seed development.

Synthesis of surface protein-imprinted nanoparticles based on metal coordination and anchored carbon dots for enhanced fluorescence detection.

Molecularly imprinted polymers endowed with photo-luminescent properties have attracted wide research interest in many fields such as biological analysis and diseases diagnosis. Herein, we illustrate a versatile method for the construction of surface protein-imprinted nanoparticles based on metal coordination and anchored carbon dots (CDs) for enhanced fluorescence detection of the target protein. As the fluorescent nanosupports for surface imprinting, CDs-attached SiO2 nanoparticles were synthesized via thiol-ene click chemistry. With histidine (His)-exposed protein as templates, imprinted nanoshells were formed over the nanosupports via copolymerization of a Cu2+-chelating monomer and an oligo (ethylene glycol) monomer, hence producing high-quality imprinted cavities because of both the relatively strong coordination and inhibited non-specific binding. Using lysozyme as a model His-exposed template, the imprinted nanoparticles showed fluorescence enhancement while binding the target protein, and exhibited significantly increased specific fluorescence response than the controls without the metal coordination. They achieved a high imprinting factor of 5.8 and a low limit of detection of 10.1 nM. Furthermore, such sensors were applied to determine lysozyme in diluted chicken egg-white samples with satisfactory recoveries at three spiking levels ranging from 97.9 to 101.4%. Human serum albumin was also used as another template protein for preliminary confirming the generality of the presented strategy.

Geographic Variation in the Petiole-Lamina Relationship of 325 Eastern Qinghai-Tibetan Woody Species: Analysis in Three Dimensions.

The petiole-lamina relationship is central to the functional tradeoff between photosynthetic efficiency and the support/protection cost. Understanding environmental gradients in the relationship and its underlying mechanisms remains a critical challenge for ecologists. We investigated the possible scaling of the petiole-lamina relationships in three dimensions, i.e., petiole length (PL) vs. lamina length (LL), petiole cross sectional area (PCA) vs. lamina area (LA), and petiole mass (PM) vs. lamina mass (LM), for 325 Qinghai-Tibetan woody species, and examined their relation to leaf form, altitude, climate, and vegetation types. Both crossspecies analysis and meta-analysis showed significantly isometric, negatively allometric, and positively allometric scaling of the petiole-lamina relationships in the length, area, and mass dimensions, respectively, reflecting an equal, slower, and faster variation in the petiole than in the lamina in these trait dimensions. Along altitudinal gradients, the effect size of the petiole-lamina relationship decreased in the length and mass dimensions but increased in the area dimension, suggesting the importance of enhancing leaf light-interception and nutrient transport efficiency in the warm zones in petiole development, but enhancing leaf support/protection in the cold zones. The significant additional influences of LA, LM, and LA were observed on the PL-LL, PCA-LA, and PM-LM relationships, respectively, implying that the single-dimension petiole trait is affected simultaneously by multidimensional lamina traits. Relative to simple-leaved species, the presence of petiolule in compound-leaved species can increase both leaf light interception and static gravity loads or dynamic drag forces on the petiole, leading to lower dependence of PL variation on LL variation, but higher biomass allocation to the petiole. Our study highlights the need for multidimension analyses of the petiole-lamina relationships and illustrates the importance of plant functional tradeoffs and the change in the tradeoffs along environmental gradients in determining the relationships.

Response of Leaf Traits of Eastern Qinghai-Tibetan Broad-Leaved Woody Plants to Climatic Factors.

Plant ecologists have long been interested in quantifying how leaf traits vary with climate factors, but there is a paucity of knowledge on these relationships given a large number of the relevant leaf traits and climate factors to be considered. We examined the responses of 11 leaf traits (including leaf morphology, stomatal structure and chemical properties) to eight common climate factors for 340 eastern Qinghai-Tibetan woody species. We showed temperature as the strongest predictor of leaf size and shape, stomatal size and form, and leaf nitrogen and phosphorus concentrations, implying the important role of local heat quantity in determining the variation in the cell- or organ-level leaf morphology and leaf biochemical properties. The effects of moisture-related climate factors (including precipitation and humidity) on leaf growth were mainly through variability in leaf traits (e.g., specific leaf area and stomatal density) related to plant water-use physiological processes. In contrast, sunshine hours affected mainly cell- and organ-level leaf size and shape, with plants developing small/narrow leaves and stomata to decrease leaf damage and water loss under prolonged solar radiation. Moreover, two sets of significant leaf trait-climate relationships, i.e., the leaf/stomata size traits co-varying with temperature, and the water use-related leaf traits co-varying with precipitation, were obtained when analyzing multi-trait relationships, suggesting these traits as good indicators of climate gradients. Our findings contributed evidence to enhance understanding of the regional patterns in leaf trait variation and its environmental determinants.

A method for synthesis of oriented epitope-imprinted open-mouthed polymer nanocapsules and their use for fluorescent sensing of target protein.

Epitope imprinting has proved to be an effective way for fabricating artificial receptors for protein recognition. Surface imprinting over sacrificial supports is particularly favorable for generating high-quality epitope-imprinted cavities, but obtaining nanomaterials by this way is still a challenge. Herein, we propose a method for the synthesis of oriented surface epitope-imprinted open-mouthed polymer nanocapsules (OM-MIP NCs) by sacrificing asymmetric template-modified Janus nanocores. Amine/aldehyde functionalized SiO2 Janus nanoparticles were prepared via the molten-wax-in-water Pickering emulsion approach, an easy scale-up technique. Epitope templates and vinyl groups were coupled to the aldehyde-bearing major side, whereas polyethylene glycol (PEG) chains were grafted to the amine-modified side. Incomplete imprinted shells were then generated principally on the non-PEGylated side via aqueous precipitation polymerization, hence affording OM-MIP NCs after etching the SiO2 nanocores. With a C-terminus nonapeptide of bovine serum albumin (BSA) chosen as a model epitope and polymerizable carbon dots added to the pre-polymerization solution, fluorescent OM-MIP NCs were synthesized for sensing of BSA. Such NCs reached maximal fluorescent response within 15 min, greatly faster than the closed imprinted NCs within 130 min, proving good accessibility of their inner-surface imprinted cavities thanks to the open mouths. Furthermore, they showed excellent target protein detection performance, with an imprinting factor of 7.8, a limit of detection of 43.8 nM and a linear range of 0.2-6 µM. The recoveries in bovine serum samples at four spiking levels ranged from 99.2 to 107.2%, with relative standard deviations of 1.2-5.9%.

Effect of Nitrogen Addition on Selection of Germination Trait in an Alpine Meadow on the Tibet Plateau.

Seed germination requirements may determine the kinds of habitat in which plants can survive. We tested the hypothesis that nitrogen (N) addition can change seed germination trait-environmental filter interactions and ultimately redistribute seed germination traits in alpine meadows. We determined the role of N addition on germination trait selection in an alpine meadow after N addition by combining a 3-year N addition experiment in an alpine meadow and laboratory germination experiments. At the species level, germination percentage, germination rate (speed) and breadth of temperature niche for germination (BTN) were positively related to survival of a species in the fertilized community. In addition, community-weighted means of germination percentage, germination rate, germination response to alternating temperature and BTN increased. However, germination response to wet-cold storage (cold stratification) and functional richness of germination traits was lower in alpine meadows with high-nitrogen addition than in those with no, low and medium N addition. Thus, N addition had a significant influence on environmental filter-germination trait interactions and generated a different set of germination traits in the alpine meadow. Further, the effect of N addition on germination trait selection by environmental filters was amount-dependent. Low and medium levels of N addition had less effect on redistribution of germination traits than the high level.

Soil abiotic properties and plant functional diversity co-regulate the impacts of nitrogen addition on ecosystem multifunctionality in an alpine meadow.

Soil abiotic properties and plant diversity have been shown to affect ecosystem functions in alpine meadow ecosystems. However, we know little about the relative importance of these factors in driving the responses of multiple ecosystem functions simultaneously (multifunctionality) to nitrogen (N) enrichment. Here, we measured soil abiotic properties (soil pH; available nitrogen, AN; available phosphorous, AP; and dissolved organic carbon, DOC) and multiple plant diversity metrics (species diversity, SD; functional diversity, FD; and phylogenetic diversity, PD) after a 5-year N fertilization experiment (0, 5, 10, and 15 g N m-2 yr-1) to evaluate their roles in mediating the impacts of N addition on aboveground plant functions (APF), soil microbial functions (SMF), and ecosystem multifunctionality (EMF) in a N-limited Tibetan alpine meadow. We found that N addition decreased APF but increased SMF and EMF. Structural equation models (SEMs) showed that APF was co-driven by soil DOC and the community weighted mean for plant traits (CWMs), and SMF was driven by soil DOC, soil AN, CWMs and functional dispersion (FDis). The effects of N addition on EMF were driven by soil AN and FDis. Our results suggest that the effects of N enrichment on APF, SMF, and EMF are driven by differential mechanisms. Furthermore, the findings suggest that FD is superior to SD and PD in mediating the responses of ecosystem functions to N enrichment.

Hyponastic Leaves 1 protects pri-miRNAs from nuclear exosome attack.

Biogenesis of plant microRNAs (miRNAs) takes place in nuclear dicing bodies (D-bodies), where the ribonulease III-type enzyme Dicer-like 1 (DCL1) processes primary transcripts of miRNAs (pri-miRNAs) into miRNA/miRNA* (*, passenger strand) duplexes from either base-to-loop or loop-to-base directions. Hyponastic Leaves 1 (HYL1), a double-stranded RNA-binding protein, is crucial for efficient and accurate processing. However, whether HYL1 has additional function remains unknown. Here, we report that HYL1 plays a noncanonical role in protecting pri-miRNAs from nuclear exosome attack in addition to ensuring processing. Loss of functions in SOP1 or HEN2, two cofactors of the nucleoplasmic exosome, significantly suppressed the morphological phenotypes of hyl1-2 Remarkably, mature miRNAs generated from loop-to-base processing were partially but preferentially restored in the hyl1 sop1 and hyl1 hen2 double mutants. Accordingly, loop-to-base-processed pri-miRNAs accumulated to higher levels in double mutants. In addition, dysfunction of HEN2, but not of SOP1, in hyl1-2 resulted in overaccumulation of many base-to-loop-processed pri-miRNAs, with most of their respective miRNAs unaffected. In summary, our findings reveal an antagonistic action of exosome in pri-miRNA biogenesis and uncover dual roles of HYL1 in stabilizing and processing of pri-miRNAs.

Transcriptome analysis reveals insight into molecular hydrogen-induced cadmium tolerance in alfalfa: the prominent role of sulfur and (homo)glutathione metabolism.

BACKGROUND: Hydrogen gas (H2) is hypothesised to play a role in plants that are coping with stresses by regulating signal transduction and gene expression. Although the beneficial role of H2 in plant tolerance to cadmium (Cd) has been investigated previously, the corresponding mechanism has not been elucidated. In this report, the transcriptomes of alfalfa seedling roots under Cd and/or hydrogen-rich water (HRW) treatment were first analysed. Then, the sulfur metabolism pathways were focused on and further investigated by pharmacological and genetic approaches. RESULTS: A total of 1968 differentially expressed genes (DEGs) in alfalfa seedling roots under Cd and/or HRW treatment were identified by RNA-Seq. The DEGs were classified into many clusters, including glutathione (GSH) metabolism, oxidative stress, and ATP-binding cassette (ABC) transporters. The results validated by RT-qPCR showed that the levels of relevant genes involved in sulfur metabolism were enhanced by HRW under Cd treatment, especially the genes involved in (homo)glutathione metabolism. Additional experiments carried out with a glutathione synthesis inhibitor and Arabidopsis thaliana cad2-1 mutant plants suggested the prominent role of glutathione in HRW-induced Cd tolerance. These results were in accordance with the effects of HRW on the contents of (homo)glutathione and (homo)phytochelatins and in alleviating oxidative stress under Cd stress. In addition, the HRW-induced alleviation of Cd toxicity might also be caused by a decrease in available Cd in seedling roots, achieved through ABC transporter-mediated secretion. CONCLUSIONS: Taken together, the results of our study indicate that H2 regulated the expression of genes relevant to sulfur and glutathione metabolism and enhanced glutathione metabolism which resulted in Cd tolerance by activating antioxidation and Cd chelation. These results may help to elucidate the mechanism governing H2-induced Cd tolerance in alfalfa.

Analyses of functional conservation and divergence reveal requirement of bHLH010/089/091 for pollen development at elevated temperature in Arabidopsis.

The Arabidopsis bHLH010/089/091 (basic helix-loop-helix) genes are functionally redundant and are required for both anther development and normal expression of DYT1-activated anther-related genes. These three genes are conserved in Brassicaceae, suggesting that each of them is under selection pressure; however, little is known about the possible functional differences among these bHLH genes and between the bHLH and DYT1 genes. Here, we compared novel anther transcriptomic data sets from bHLH010/089/091 single and double mutants, with an anther transcriptomic data set from the wild type (WT) and a previously obtained anther transcriptomic data set from the bhlh010 bhlh089 bhlh091 triple mutant. The results revealed molecular phenotypes that support the functional redundancy and divergence of bHLH010, bHLH089, and bHLH091, as well as the functional overlap and difference between them and DYT1. DNA-binding analyses revealed that DYT1 and bHLH089 specifically recognize the TCATGTGC box to activate the expression of target genes, including ATA20, EXL4, and MEE48. In addition, among genes whose expression was affected in the bhlh010 bhlh089 double and bhlh010 bhlh089 bhlh091 triple mutants, genes that are involved in the stress response and cell signaling were enriched, which included 256 genes whose expression was preferentially induced by heat during early flower development. Moreover, the bhlh double mutants exhibited defective pollen development when the plants were grown under elevated temperature, suggesting that bHLH genes are important for anther gene expression under such conditions. These results are consistent with the observation that the heat-induced expression of several genes is less in the bhlh mutants than that in the WT. Therefore, our results provide important insights into the molecular mechanism underlying the activation of direct targets by DYT1-bHLH089 heterodimers and demonstrate the protective roles of bHLH010/089/091 in maintaining fertility upon heat stress.