Transcription factors RhbZIP17 and RhWRKY30 enhance resistance to Botrytis cinerea by increasing lignin content in rose petals.

The petals of ornamental plants such as roses (Rosa spp.) are the most economically important organs. This delicate, short-lived plant tissue is highly susceptible to pathogens, in large part because the walls of petal cells are typically thinner and more flexible compared with leaf cells, allowing the petals to fold and bend without breaking. The cell wall is a dynamic structure that rapidly alters its composition in response to pathogen infection, thereby reinforcing its stability and boosting plant resistance against diseases. However, little is known about how dynamic changes in the cell wall contribute to resistance to Botrytis cinerea in rose petals. Here, we show that the B. cinerea-induced transcription factor RhbZIP17 is required for the defense response of rose petals. RhbZIP17 is associated with phenylpropanoid biosynthesis and binds to the promoter of the lignin biosynthesis gene RhCAD1, activating its expression. Lignin content showed a significant increase under gray mold infection compared with the control. RhCAD1 functions in the metabolic regulation of lignin production and, consequently, disease resistance, as revealed by transient silencing and overexpression in rose petals. The WRKY transcription factor RhWRKY30 is also required to activate RhCAD1 expression and enhance resistance against B. cinerea. We propose that RhbZIP17 and RhWRKY30 increase lignin biosynthesis, improve the resistance of rose petals to B. cinerea, and regulate RhCAD1 expression.

Estimate of the degradation potentials of cellulose, xylan, and chitin across global prokaryotic communities.

Complex polysaccharides (e.g. cellulose, xylan, and chitin), the most abundant renewable biomass resources available on Earth, are mainly degraded by microorganisms in nature. However, little is known about the global distribution of the enzymes and microorganisms responsible for the degradation of cellulose, xylan, and chitin in natural environments. Through large-scale alignments between the sequences released by the Earth Microbiome Project and sequenced prokaryotic genomes, we determined that almost all prokaryotic communities have the functional potentials to degrade cellulose, xylan, and chitin. The median abundances of genes encoding putative cellulases, xylanases, and chitinases in global prokaryotic communities are 0.51 (0.17-1.01), 0.24 (0.05-0.57), and 0.33 (0.11-0.71) genes/cell, respectively, and the composition and abundance of these enzyme systems are environmentally varied. The taxonomic sources of the three enzymes are highly diverse within prokaryotic communities, and the main factor influencing the diversity is the community's alpha diversity index rather than gene abundance. Moreover, there are obvious differences in taxonomic sources among different communities, and most genera with degradation potentials are narrowly distributed. In conclusion, our analysis preliminarily depicts a panorama of cellulose-, xylan-, and chitin-degrading enzymatic systems across global prokaryotic communities.

Solid-phase extraction based on PDMS/ionic liquid sponge followed by gas chromatography-mass spectrometry for rapid and sensitive determination of volatile components in lavender.

INTRODUCTION: Due to the complexity and the low concentrations of volatile components in aromatic plants, sample pretreatment is an important step in the whole analytical procedure. OBJECTIVE: This study aimed to propose a novel, sensitive and fast approach to determine the volatile components in lavender. METHODS: The 1-butyl-3-(propyltrimethoxysilane)imidazolium chloride ([BPtmsim]Cl) ionic liquid was introduced onto the surface of polydimethylsiloxane (PDMS) to prepare a novel PDMS/[BPtmsim]Cl sponge with large surface area, good sorption performance, and reusability. A solid-phase extraction method was developed based on PDMS/[BPtmsim]Cl sponge combined with gas chromatography-mass spectrometry (GC-MS). RESULTS: The effects of the various experimental parameters on the extraction efficiency were investigated. The optimal conditions were [BPtmsim]Cl amount of 0.3 g, 1:4 as the mass ratio of PDMS/[BPtmsim]Cl to lavender sample, microwave power of 700 W, microwave time of 10 min, and n-hexane as the desorption solvent. The method validation results showed good linearity (10-800 µg/ml), high correlation coefficient (R2 ≥ 0.9991), low limits of detection (1.73-2.50 ng/µl), and limits of quantification (4.10-5.11 ng/µl). The interday and intraday precision with relative standard deviation (RSD) values were below 1.93% and 4.71%, respectively. Under the optimal extraction conditions, 16 lavender samples from three different species were analysed and a total of 57 volatile compounds were identified. The correlation between different species of lavender and volatile components was explored using principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA). CONCLUSION: The results showed that PDMS/[BPtmsim]Cl extraction is a rapid, highly efficient, and sensitive technique for the determination of volatile components in complex plant samples.

PuC3H35 confers drought tolerance by enhancing lignin and proanthocyanidin biosynthesis in the roots of Populus ussuriensis.

Since the roots are the very organ where plants first sense and respond drought stress, it is of great importance to better understand root responses to drought. Yet the underlying molecular mechanisms governing root responses to drought stress have been poorly understood. Here, we identified and functionally characterized a CCCH type transcription factor, PuC3H35, and its targets, anthocyanin reductase (PuANR) and early Arabidopsis aluminum induced1 (PuEARLI1), which are involved in mediating proanthocyanidin (PA) and lignin biosynthesis in response to drought stress in Populus ussuriensis root. PuC3H35 was root-specifically induced upon drought stress. Overexpressing PuC3H35 promoted PA and lignin biosynthesis and vascular tissue development, resulting in enhanced tolerance to drought stress by the means of anti-oxidation and mechanical supporting. We further demonstrated that PuC3H35 directly bound to the promoters of PuANR and PuEARLI1 and overexpressing PuANR or PuEARLI1 increased root PA or lignin levels, respectively, under drought stress. Taken together, these results revealed a novel regulatory pathway for drought tolerance, in which PuC3H35 mediated PA and lignin biosynthesis by collaboratively regulating 'PuC3H35-PuANR-PA' and 'PuC3H35-PuEARLI1-PuCCRs-lignin' modules in poplar roots.

Polydimethylsiloxane/graphene oxide/ß-cyclodextrin sponge as a solid-phase extraction sorbent coupled with gas chromatography-mass spectrometry for rapid adsorption and sensitive determination of lavender essential oil.

An adsorbent polydimethylsiloxane/graphene oxide/ß-cyclodextrin sponge, which possessed the merits of high surface area, chemical stability, environment friendly, and excellent extraction capacity, was successfully fabricated. Based on the advantages, a novel microwave-assisted headspace solid-phase extraction method for lavender essential oil using polydimethylsiloxane/graphene oxide/ß-cyclodextrin sponge as adsorbents was developed in this study. Various experimental parameters were studied. The optimal extraction conditions were as follows: 1 mg/mL as dopamine solution concentration, graphene oxide dosages of 30 mg, microwave power of 700 W, microwave irradiation time of 10 min, and desorption solvent of n-hexane. Under the optimal extraction condition, linearities ranging from 10 to 800 ng were achieved for six representative compounds with a correlation coefficients value of >0.99. The intra-day and inter-day precisions were in the ranges of 0.40-1.56 and 0.67-2.56%, respectively. Finally, the proposed technique was applied to analyze essential oil constituents in 14 samples of three lavender varieties, and 48 compounds were identified. Lavender varieties were distinguished using principal component analysis and partial least squares discriminant analysis. The results showed that the method developed in this study is a novel, simple, and sensitive method for the determination of essential oil in complex plant samples.

Skeletal Class III Malocclusion Is Associated with ADAMTS2 Variants and Reduced Expression in a Familial Case.

Skeletal Class III malocclusion with maxillary deficiency is a severe maxillofacial disease with unclear pathogenic mechanisms. We recruited a Han Chinese family who was clinically diagnosed with skeletal Class III malocclusion and maxillary deficiency. Using whole exome sequencing, a missense variant in ADAMTS2 (NM_014244: c.3506G>T: p.G1169V) was identified and predicted as deleterious by in silico tools. We also found ADAMTS2 variants associated with deficient maxillary development in a cohort. ADAMTS2 expression in HEK293 cells showed significant decrease due to the variant, which was also consistent in dental pulp stem cells from the proband and a healthy control. In the adamts2-knockdown zebrafish model, the length and width of the ethmoid plate, as well as the length of the palatoquadrate became significantly shorter than the control group (p < 0.001), while there was no significant difference in the length and width of the mandible. The expression of Sox3, which was required in early embryonic craniofacial development, was significantly downregulated in the adamts2-knockdown zebrafish embryos. Bioinformatic and cellular studies showed that the decreased expression of ADAMTS2 may inhibit downstream ErbB signaling pathway transduction and restrain subsequent osteogenesis in human adult mesenchymal stromal cells. Collectively, these data showed that ADAMTS2 (c.3506G>T: p.G1169V) may confer susceptibility to risk of skeletal Class III malocclusion with maxillary deficiency.

Persistent pollution episodes, transport pathways, and potential sources of air pollution during the heating season of 2016-2017 in Lanzhou, China.

As one of the most important industrial cities in Northwest China, Lanzhou currently suffers from serious air pollution. This study analyzed the formation mechanism and potential source areas of persistent air pollution in Lanzhou during the heating period from November 1, 2016 to March 31, 2017 based on the air pollutant concentrations and relevant meteorological data. Our findings indicate that particulate pollution was extremely severe during the study period. The daily PM2.5 and PM10 concentrations had significantly negative correlations with daily temperature, wind speed, maximum daily boundary layer height, while the daily PM2.5 and PM10 concentrations showed significantly positive correlations with daily relative humidity. Five persistent pollution episodes were identified and classified as either stagnant accumulation or explosive growth types according to the mechanism of pollution formation and evolution. The PM2.5 and PM10 concentrations and PM2.5/PM10 ratio followed a growing "saw-tooth cycle" pattern during the stagnant accumulation type event. Dust storms caused abrupt peaks in PM10 and a sharp decrease in the PM2.5/PM10 ratio in explosive growth type events. The potential sources of PM10 were mainly distributed in the Kumtag Desert in Xinjiang Uygur Autonomous Region, the Qaidam Basin and Hehuang Valley in Qinghai Province, and the western and eastern Hexi Corridor in Gansu Province. The contributions to PM10 were more than 120 µg/m3. The important potential sources of PM2.5 were located in Hehuang Valley in Qinghai and Linxia Hui Autonomous Prefecture in Gansu; the concentrations of PM2.5 were more than 60 µg/m3.

Effects of alkalinity on interaction between EPS and hydroxy-aluminum with different speciation in wastewater sludge conditioning with aluminum based inorganic polymer flocculant.

Extracellular polymeric substances (EPS) form a stable gel-like structure to combine with water molecules through steric hindrance, making the mechanical dewatering of wastewater sludge considerably difficult. Coagulation/flocculation has been widely applied in improving the sludge dewatering performance, while sludge properties (organic fraction and solution chemistry conditions) are highly changeable and have important effects on sludge flocculation process. In this work, the alkalinity effects on sludge conditioning with hydroxy-aluminum were comprehensively investigated, and the interaction mechanisms between EPS and hydroxy-aluminum with different speciation were unraveled. The results showed that the effectiveness of hydroxy-aluminum conditioning gradually deteriorated with increase in alkalinity. Meanwhile, the polymeric hydroxy-aluminum (Al13) and highly polymerized hydroxy-aluminum (Al30) were hydrolysed and converted into amorphous aluminum hydroxide (Al(OH)3), which changed the flocculation mechanism from charge neutralization and complexing adsorption to hydrogen bond interaction. Additionally, both Al13 and Al30 showed higher binding capacity for proteins and polysaccharides in EPS than monomeric aluminum and Al(OH)3. Al13 and Al30 coagulation changed the secondary structure of proteins in EPS, which caused a gelation reaction to increase molecular hydrophobicity of proteins and consequently sludge dewaterability. This study provided a guidance for optimizing the hydroxy-aluminum flocculation conditioning of sludge with high solution alkalinity.

Electrodeposited poly(3,4-ethylenedioxythiophene) doped with graphene oxide for the simultaneous voltammetric determination of ascorbic acid, dopamine and uric acid.

Poly(3,4-ethylenedioxythiophene) (PEDOT) films were electrodeposited by cyclic voltammetry on a glassy carbon electrode (GCE) in aqueous solution. Three kinds of supporting electrolytes were used, viz. graphene oxide (GO), phosphate buffered saline (PBS), and GO in PBS, respectively. The surface morphology of the modified electrodes was characterized by scanning electron microscopy. The electrochemical performance of the modified electrodes was investigated by cyclic voltammetry and electrochemical impedance spectroscopy by using the hexacyanoferrate redox system. The results demonstrate that the PEDOT-GO/GCE, which was electropolymerized in aqueous solutions containing EDOT and GO, shows the best electrochemical activities compared with other modified electrodes. The electrochemical behaviors of ascorbic acid (AA), dopamine (DA) and uric acid (UA) were investigated by cyclic voltammetry. The PEDOT-GO/GCE exhibits enhanced electrocatalytic activities towards these important biomolecules. Under physiological pH conditions and in the mixed system of AA, DA and UA, the modified GCE exhibits the following figures of merit: (a) a linear voltammetric response in the concentration ranges of 100-1000 µM for AA, 6.0-200 µM for DA, and 40-240 µM for UA; (b) well separated oxidation peaks near 31, 213 and 342 mV (vs. saturated Ag/AgCl) for AA, DA and UA, respectively; and (c) detection of limits (at S/N = 3) of 20, 2.0 and 10 µM. The results demonstrate that GO, based on its relatively large number of anionic sites, can be used as the sole weak electrolyte and charge balance dopant for the preparation of functionally doped conducting polymers by electrodeposition. Graphical abstractSchematic representation of a nanostructure composed of hybrid conducting polymer PEDOT-GO nanocomposites, and its application to simultaneous determination of ascorbic acid, dopamine and uric acid.

Acyl-CoA synthetases from Physcomitrella, rice and Arabidopsis: different substrate preferences but common regulation by MS188 in sporopollenin synthesis.

MAIN CONCLUSION: ACOS5, OsACOS12 and PpACOS6 are all capable of fatty acyl-CoA synthetase activity but exhibit different substrate preferences. The transcriptional regulation of ACOS for sporopollenin synthesis appears to have been conserved in Physcomitrella, rice and Arabidopsis during evolution. Sporopollenin is the major constituent of spore and pollen exines. In Arabidopsis, acyl-CoA synthetase 5 (ACOS5) is an essential enzyme for sporopollenin synthesis, and its orthologues are PpACOS6 from the moss Physcomitrella and OsACOS12 from monocot rice. However, knowledge regarding the evolutionary conservation and divergence of the ACOS gene in sporopollenin synthesis remains limited. In this study, we analysed the function and regulation of PpACOS6 and OsACOS12. A complementation test showed that OsACOS12 driven by the ACOS5 promoter could partially restore the male fertility of the acos5 mutant in Arabidopsis, while PpACOS6 did not rescue the acos5 phenotype. ACOS5, PpACOS6 and OsACOS12 all complemented the acyl-CoA synthetase-deficient yeast strain (YB525) phenotype, although they exhibited different substrate preferences. To understand the conservation of sporopollenin synthesis regulation, we constructed two constructs with ACOS5 driven by the OsACOS12 or PpACOS6 promoter. Both constructs could restore the fertility of acos5 plants. The MYB transcription factor MS188 from Arabidopsis directly regulates ACOS5. We found that MS188 could also bind the promoters of OsACOS12 and PpACOS6 and activate the genes driven by the promoters, suggesting that the transcriptional regulation of these genes was similar to that of ACOS5. These results show that the ACOS gene promoter region from Physcomitrella, rice and Arabidopsis has been functionally conserved during evolution, while the chain lengths of fatty acid-derived monomers of sporopollenin vary in different plant species.

Non-syndromic cleft lip with or without palate-susceptible SNPs is associated with hyperdontia.

BACKGROUND: Non-syndromic supernumerary teeth (NSST) or hyperdontia may share common genetic determinants with non-syndromic cleft lip with or without palate (NSCL/P). The aim of this study was to test the associations between five genome-wide-associated NSCL/P-susceptible single nucleotide polymorphisms (SNPs) (rs2235371, rs7078160, rs8049367, rs4791774, and rs13041247) and the occurrence of NSST. MATERIALS AND METHODS: A total of 163 cases and 326 controls were recruited and their genomic DNA was extracted from blood samples. Five NSCL/P-susceptible SNPs (rs2235371, rs7078160, rs8049367, rs4791774, and rs13041247) were genotyped by TaqMan method. Odds ratio (OR) and 95% confidence interval (CI) were used to estimate the associations between the SNPs and the risk of NSST by PLINK software. RESULTS: Rs4791774 (A > G) and rs13041247 (T > C) were associated with risk of NSST (rs4791774: Padd  = 0.011, OR, 95% CI = 0.62, 0.43-0.90; rs13041247: Phomo  = 0.031, OR, 95% CI = 1.79, 1.05-3.05) and one supernumerary tooth (rs4791774: Pdom  = 0.009, OR, 95% CI = 0.56, 0.36-0.87; rs13041247: Phomo  = 0.034, OR, 95% CI = 1.82, 1.05-3.15). Rs4791774 (A > G) was also showed association with risk of upper arch supernumerary teeth only (Padd  = 0.010, OR, 95% CI = 0.60, 0.41-0.89). CONCLUSION: Non-syndromic cleft lip with or without palate-susceptible loci rs4791774 (A > G) and rs13041247 (T > C) were associated with the risk of supernumerary teeth.

Non-syndromic cleft lip with or without palate susceptible loci is associated with tooth agenesis.

OBJECTIVE: Non-syndromic tooth agenesis (NSTA) may share common genetic factors with non-syndromic cleft lip with or without cleft palate (NSCL/P). Single-nucleotide polymorphisms (SNPs) were associated with individual's susceptibility to these anomalies. We selected five NSCL/P-associated SNPs from our previous genome-wide association study (GWAS) to test for the associations with NSTA. MATERIALS AND METHODS: A total of 677 NSTA cases and 1,144 healthy controls were recruited in this case-control study. Five genome-wide NSCL/P-associated SNPs (rs2235371, rs7078160, rs8049367, rs4791774, and rs13041247) were genotyped by TaqMan platform and evaluated for the associations with NSTA using plink software. RESULTS: No significant associations between these SNPs and risk of NSTA were observed in the overall analysis and subgroup analysis with the number of missing teeth. However, in the subgroup analysis by tooth position, rs8049367 was nominally associated with mandibular premolar agenesis (Dominant model: ORdom  = 0.66, 95% CIdom  = 0.47-0.93, pdom  = 0.016; Heterozygote model: ORhet  = 0.60, 95% CIhet  = 0.41-0.88, Phet  = 0.008). Rs4791774 showed a nominal association with congenitally missing maxillary canine (Dominant model: ORdom  = 0.53, 95% CIdom  = 0.28-0.98, pdom  = 0.041; Heterozygote model: ORhet  = 0.50, 95% CIhet  = 0.26-0.97, Phet  = 0.041) and premolar (Additive model: OR = 0.59, 95% CI = 0.36-0.96, p = 0.035). CONCLUSION: This study showed that NSCL/P susceptible loci rs8049367 and rs4791774 were probably associated with the risk of NSTA.

Characterization and cytocompatibility of thermosensitive hydrogel embedded with chitosan nanoparticles for delivery of bone morphogenetic protein-2 plasmid DNA.

A novel injectable chitosan thermosensitive hydrogel was designed as a target multi-effect scaffold for endogenous repair of the periodontium. The hydrogel complex was designed by embedding chitosan nanoparticles (CSn) loaded with bone morphogenetic protein-2 plasmid DNA (pDNA-BMP2) into a chitosan (CS)-based hydrogel with α,ß-glycerophosphate (α,ß-GP), termed CS/CSn(pDNA-BMP2)-GP. Characterization, the in vitro release profile for pDNA-BMP2, and cytocompatibility to human periodontal ligament cells (HPDLCs), were then conducted. The average diameter of the CSn(pDNA-BMP2) was 270.1 nm with a polydispersity index (PDI) of 0.486 and zeta potential of +27.0 mv. A DNase I protection assay showed that CSn could protect the pDNA-BMP2 from nuclease degradation. Encapsulation efficiency and loading capacity of CSn(pDNA-BMP2) were more than 80 and 30 %, respectively. The sol-gel transition time was only 3 min when CSn(pDNA-BMP2) was added into the CS/α,ß-GP system. Scanning electron microscopy showed that CSn(pDNA-BMP2) was randomly dispersed in a network with regular holes and a porous structure. Weighting method showed the swelling ratio and degradation was faster in medium of pH 4.0 than pH 6.8. An in vitro pDNA-BMP2 release test showed that the cumulative release rate of pDNA-BMP2 was much slower from CS/CSn-GP than from CSn in identical release media. In release media with different pH, pDNA-BMP2 release was much slower at pH 6.8 than at pH 4.0. Three-dimensional culture with HPDLCs showed good cell proliferation and the Cell-Counting Kit-8 assay indicated improved cell growth with the addition of CSn(pDNA-BMP2) to CS/α,ß-GP. In summary, the CS/CSn(pDNA-BMP2)-GP complex system exhibited excellent biological properties and cytocompatibility, indicating great potential as a gene delivery carrier and tissue regeneration scaffold for endogenous repair of the periodontium.

What are patients' expectations of orthodontic treatment: a systematic review.

BACKGROUND: What patients expect to happen during treatment or benefit from the treatment might influence the subsequent factors such as treatment outcome, patient satisfaction, patient's cooperation as well as compliance. The aim of this systematic review is to assess the role of patients' expectations from orthodontic treatment. METHODS: A systematic literature search of four databases Pubmed, Cochrane, Web of Science and PsychINFO was conducted following PRISMA guidelines. Studies reporting expectations regarding orthodontic treatment were selected and a narrative review was conducted. The quality of study was rated according to STROBE statements and the methodology as well as key findings were summarized. RESULTS: Thirteen studies (14 papers) were finally included for analysis. Among them, only one was a randomized control trial, while the rest included one cohort study, two questionnaire-developments and ten cross-sectional studies. The STROBE quality of reporting scores of the studies ranged from 12 to 18. Seven papers described expectations of the treatment experiences, along with seven talking about benefit expectations from the treatment. Dental appearance and function improvement were most expected in studies relate to the treatment benefits. CONCLUSIONS: Orthodontics appears to have adopted various standardized questionnaires. However, most of them are poor in the quality of methodology and results analyses, which prohibit synthesizing sufficient evidence to help identify which factors influence patient expectations. The evidence of "expectations" affecting treatment outcomes is not found in current research. Future studies are needed to better understand the impact of "expectation" on the treatment both theoretically and experimentally.

Efficient extraction based on a polydimethylsiloxane/bimetallic ZnCo-MOF carbonization sponge coupled with GC-MS for the rapid analysis of volatile compounds in cumin.

A novel porous polydimethylsiloxane/bimetallic ZnCo-MOF carbonization (PDMS/ZnCo-MOF@C) sponge was successfully fabricated, followed by its utilization in GC-MS for the high efficiency extraction and determination of volatile compounds in cumin. The PDMS/ZnCo-MOF@C sponge exhibits outstanding properties with a considerable adsorption capacity, high surface area, and large pore volume and has shown potential as an ideal adsorbent for the separation and preconcentration of trace volatile compounds. The effect of different parameters on the extraction efficiency were investigated. Excellent analytical performances were achieved for the representative compounds (ß-pinene, p-cymene, γ-terpinene, cuminaldehyde, and linalyl acetate), including wide linearity (2.31-440.1 ng) with high correlation coefficients (R2 ≥ 0.9979), low LODs (1.02-3.11 ng) and LOQs (2.45-7.08 ng), and satisfactory precision (intra-day RSDs ≤ 2.89% and inter-day RSDs ≤ 4.14%). The optimal method was applied for the analysis of cumin from different regions and 44 volatile compounds were identified. The correlation between the different regions of cumin and volatile compounds was explored using multivariate statistical analysis. These results demonstrated that PDMS/ZnCo-MOF@C is an efficient, simple and sensitive material for use in the pretreatment technique for the determination of the volatile compounds in aromatic plants.

Limited Sampling Strategies for Estimating Busulfan Area Under the Concentration-Time Curve: Based on Peak and Trough Concentrations in Saliva.

Therapeutic drug monitoring for busulfan is currently performed by multiple plasma sampling. Saliva is considered a noninvasive therapeutic drug monitoring matrix. This study aimed to investigate intravenous busulfan pharmacokinetics (PK) in plasma and saliva, and establish a limited sampling strategy (LSS) for predicting the area under the concentration-time curve from time zero to infinity in plasma (AUC0-∞,p) by using saliva samples. Therefore, the PK of busulfan was studied in 37 Chinese patients. Pearson correlation analysis was used to evaluate the correlation between the AUC of busulfan in plasma and saliva. LSS models were established by the multiple linear regression analysis. The prediction error, the mean prediction error, and the root mean square error were used to evaluate the predictive accuracy. The agreement between the predicted and observed AUC0-∞ in saliva was investigated by the intraclass correlation coefficient and Bland-Altman analysis. The accuracy and robustness of the models were evaluated by using the bootstrap procedure. The result of PK analysis 62.2% of patients (23/37) was within the target range of AUC0-∞,p . A good correlation between saliva and plasma busulfan AUC0-∞ was observed (r = 0.63, p < .01). The bias and precision of the models 7 and 13 were less than 15%. The intraclass correlation coefficient exceeded 0.9, and the limits of agreement were within ±15%. The 2-point LSS model in saliva is a convenient and desirable approach to predict the AUC0-∞ of 4 times daily intravenous busulfan in plasma, which can be used to design personalized dosing for busulfan.

METTL3-dependent m6A modification of PSEN1 mRNA regulates craniofacial development through the Wnt/ß-catenin signaling pathway.

Craniofacial malformations, often associated with syndromes, are prevalent birth defects. Emerging evidence underscores the importance of m6A modifications in various bioprocesses such as stem cell differentiation, tissue development, and tumorigenesis. Here, in vivo, experiments with zebrafish models revealed that mettl3-knockdown embryos at 144 h postfertilization exhibited aberrant craniofacial features, including altered mouth opening, jaw dimensions, ethmoid plate, tooth formation and hypoactive behavior. Similarly, low METTL3 expression inhibited the proliferation and migration of BMSCs, HEPM cells, and DPSCs. Loss of METTL3 led to reduced mRNA m6A methylation and PSEN1 expression, impacting craniofacial phenotypes. Co-injection of mettl3 or psen1 mRNA rescued the level of Sox10 fusion protein, promoted voluntary movement, and mitigated abnormal craniofacial phenotypes induced by mettl3 knockdown in zebrafish. Mechanistically, YTHDF1 enhanced the mRNA stability of m6A-modified PSEN1, while decreased METTL3-mediated m6A methylation hindered ß-catenin binding to PSEN1, suppressing Wnt/ß-catenin signaling. Pharmacological activation of the Wnt/ß-catenin pathway partially alleviated the phenotypes of mettl3 morphant and reversed the decreases in cell proliferation and migration induced by METTL3 silencing. This study elucidates the pivotal role of METTL3 in craniofacial development via the METTL3/YTHDF1/PSEN1/ß-catenin signaling axis.

Synergy between nanoplastics and benzo[a]pyrene promotes senescence by aggravating ferroptosis and impairing mitochondria integrity in Caenorhabditis elegans.

Micro-nano plastics have been reported as important carriers of polycyclic aromatic hydrocarbons (PAHs) for long-distance migration in the environment. However, the combined toxicity from long-term chronic exposure beyond the vehicle-release mechanism remains elusive. In this study, we investigated the synergistic action of Benzo[a]pyrene (BaP) and Polystyrene nanoparticles (PS) in Caenorhabditis elegans (C. elegans) as a combined exposure model with environmental concentrations. We found that the combined exposure to BaP and PS, as opposed to single exposures at low concentrations, significantly shortened the lifespan of C. elegans, leading to the occurrence of multiple senescence phenotypes. Multi-omics data indicated that the combined exposure to BaP and PS is associated with the disruption of glutathione homeostasis. Consequently, the accumulated reactive oxygen species (ROS) cannot be effectively cleared, which is highly correlated with mitochondrial dysfunction. Moreover, the increase in ROS promoted lipid peroxidation in C. elegans and downregulated Ferritin-1 (Ftn-1), resulting in ferroptosis and ultimately accelerating the aging process of C. elegans. Collectively, our study provides a new perspective to explain the long-term compound toxicity caused by BaP and PS at real-world exposure concentrations.

Microwave-induced plasma desorption/ionization source for ambient mass spectrometry.

A new ionization source based on microwave induced plasma was developed for ambient desorption/ionization. The microwave-induced plasma desorption/ionization source (MIPDI) was composed of a copper Surfatron microwave cavity where a fused-silica tube was centered axially. Stable nonlocal thermodynamic equilibrium plasma was generated in the quartz discharge tube when a microwave at a frequency of 2450 MHz was coupled to the microwave cavity. Analytes deposited on the surface of poly(tetrafluoroethylene) (PTFE) or quartz slide after hydrofluoric acid (HF) etching were desorbed and ionized by the plasma. The performance of the MIPDI technique was validated by the analysis of a variety of chemical substances, polymer compounds, and pharmaceutical drugs using argon or helium as the discharge gas. Protonated [M + H](+) or deprotonated [M - H](-) ions were observed in the positive or negative mode. MIPDI was also used for the analysis of compounds in a complex matrix without any sample preparation. MIPDI was also capable of analyzing liquid samples. The signal-to-noise ratio was 463 in the analysis of 9.2 ng of phenylalanine, and the limit of detection was 60 pg for phenylalanine. MIPDI could desorb and ionize analytes with a molecular weight of up to 1200, which was demonstrated by the analysis of polyethylene glycol 800 (PEG800). MIPDI has advantages of simple instrumentation, relatively high temperature, stability, and reproducibility.

Genotoxicity-related chemistry of human metabolites of benzo[ghi]perylene (B[ghi]P) investigated using electro-optical arrays and DNA/microsome biocolloid reactors with LC-MS/MS.

There is limited and sometimes contradictory information about the genotoxicity of the polycyclic aromatic hydrocarbon benzo[ghi]perylene (B[ghi]P). Using recently developed metabolic toxicity screening arrays and a biocolloid reactor-LC-MS/MS approach, both featuring films of DNA and human metabolic enzymes, we demonstrated the relatively low reactivity of metabolically activated B[ghi]P toward DNA. Electro-optical toxicity screening arrays showed that B[ghi]P metabolites damage DNA at a 3-fold lower rate than benzo[a]pyrene (B[a]P), whose metabolites have a strong and well-understood propensity for DNA damage. Metabolic studies using magnetic bead biocolloid reactors coated with microsomal enzymes in 96-well plates showed that cyt P450s 1A1 and 1B1 provide high activity for B[ghi]P and B[a]P conversion. Consistent with published results, the major metabolism of B[ghi]P involved oxidations at 3,4 and 11,12 positions, leading to the formation of B[ghi]P 3,4-oxide and B[ghi]P 3,4,11,12-bisoxide. B[ghi]P 3,4-oxide was synthesized and reacted with deoxyadenosine at N6 and N7 positions and with deoxyguanosine at the N2 position. B[ghi]P 3,4-oxide is hydrolytically unstable and transforms into the 3,4-diol or converts to 3- or 4-hydroxy B[ghi]P. LC-MS/MS of reaction products from the magnetic biocolloid reactor particles coated with DNA and human enzymes revealed for the first time that a major DNA adduct results from the reaction between B[ghi]P 3,4,11,12-bisoxide and deoxyguanosine. Results also demonstrated 5-fold lower formation rates of the major DNA adduct for B[ghi]P metabolites compared to B[a]P. Overall, results from both the electro-optical array and biocolloid reactor-LC-MS/MS consistently suggest a lower human genotoxicity profile of B[ghi]P than B[a]P.