Circulating small extracellular vesicle-based miRNA classifier for follicular thyroid carcinoma: a diagnostic study.

BACKGROUND: The diagnosis of follicular thyroid carcinoma (FTC) prior to surgery remains a major challenge in the clinic. METHODS: This multicentre diagnostic study involved 41 and 150 age- and sex-matched patients in the training cohort and validation cohort, respectively. The diagnostic properties of circulating small extracellular vesicle (sEV)-associated and cell-free RNAs were compared by RNA sequencing in the training cohort. Subsequently, using a quantitative real-time polymerase chain reaction (qRT‒PCR) assay, high-quality candidates were identified to construct an RNA classifier for FTC and verified in the validation cohort. The parallel expression, stability and influence of the RNA classifier on surgical strategy were also investigated. RESULTS: The diagnostic properties of sEV long RNAs, cell-free long RNAs and sEV microRNAs (miRNAs) were comparable and superior to those of cell-free miRNAs in RNA sequencing. Given the clinical application, the circulating sEV miRNA (CirsEV-miR) classifier was developed from five miRNAs based on qRT‒PCR data, which could well identify FTC patients (area under curve [AUC] of 0.924 in the training cohort and 0.844 in the multicentre validation cohort). Further tests revealed that the CirsEV-miR score was significantly correlated with the tumour burden, and the levels of sEV miRNAs were also higher in sEVs from the FTC cell line, organoid and tissue. Additionally, circulating sEV miRNAs remained constant after different treatments, and the addition of the CirsEV-miR classifier as a biomarker improves the current surgical strategy. CONCLUSIONS: The CirsEV-miR classifier could serve as a noninvasive, convenient, specific and stable auxiliary test to help diagnose FTC following ultrasonography.

Posterior capsular radial sign: a novel method to confirm anterior vitreous cortex resection in phacovitrectomy.

BACKGROUND: The main purpose of this paper is to introduce a method that can accurately locate the posterior capsule of the lens to facilitate a relatively complete resection of the anterior vitreous body. METHODS: A total of 51 patients in the experimental group and control group were enrolled in this study. Phacoemulsification combined with vitrectomy was performed in all cases. After the cataract procedure was completed in the control group, the surgeon performed a conventional anterior vitrectomy with the operative eye. In the experimental group, anterior vitrectomy was performed according to the threadiness corrugation of the posterior capsule of the lens. During the operation, with the help of triamcinolone, two surgeons confirmed the resection of the anterior vitreous cortex; the best corrected visual acuity and intraocular pressure of all patients were recorded at 1 week, 1 month and 3 months after surgery. RESULTS: Fifty patients underwent phacoemulsification combined with vitrectomy, except one patient in the experimental group who was lost to follow-up. After surgery, no significant complications were observed in all patients except two patients in the control group with temporary increases in intraocular pressure. There was no significant difference in preoperative visual acuity between the two groups (t = 0.83, P = 0.25). Both groups had varying degrees of improvement in best corrected visual acuity at 1 week, 1 month and 3 months after surgery. Moreover, there was no significant difference in BCVA between the two groups at the three follow-up time points (t=-1.15, -1.65, -1.09, P = 0.53, 0.21, 0.23). After surgery, no significant complications were observed in all patients except two patients in the control group with temporary increases in intraocular pressure. Incomplete resection of the anterior vitreous cortex was observed in 2 patients in each group, but there was no significant difference (χ2 = 7.81, P > 0.05). CONCLUSION: In the process of cataract surgery combined with vitrectomy, thready corrugation appears in the posterior capsule of the lens and is an important sign of its localization. Anterior vitrectomy can be accomplished safely and effectively with the help of thread-like corrugation, and the surgical effect is almost the same as that of traditional surgery. Especially suitable for beginners in vitreous surgery.

Lack of Interaction Motivation in Older Adults Automatically Reduces Cognitive Empathy.

Empathy, the ability to understand and respond to the experiences of others, is an important skill for maintaining good relationships throughout one's life. Previous research indicated that emotional empathy remained stable or even increased in older adults compared to younger adults, while cognitive empathy showed age-related deficits. Based on the selective engagement hypothesis, this deficit was not caused by a decline in cognitive functioning, but by a lack of willingness to judge the target person's emotions more precisely, that is, by a lack of interaction motivation. In order to provide more evidence on the causes of empathic aging in older adults, the current study investigated the influence of interaction motivation on empathy in older adults in an Eastern cultural context (China) based on the selective engagement hypothesis. This study used older adults and younger adults as subjects. Through two experiments, empathy was measured by the multiple empathy test (Experiment 1) and film tasks (Experiment 2); at the same time, use accountability instructions (Experiment 1), the age-related events (Experiment 2) to manipulate interaction motivation. The results showed that emotional empathy was significantly higher in older adults than in younger adults, regardless of whether interaction motivation was elicited. In terms of cognitive empathy, when there is no motivation, the cognitive empathy of older adults is significantly lower than that of younger adults. When the interaction motivation is stimulated, the cognitive empathy of older adults is no less than that of younger adults. This suggested that empathic aging in older adults was not a permanent decline in cognitive empathy, but rather a decline in interaction motivation, supporting the selective engagement hypothesis.

Socioeconomic development shows positive links to the conservation efficiency of China's protected area network.

While the protected area (PA) covers >15% of the planet's terrestrial land area and continues to expand, factors determining its effectiveness in conserving endangered species are being debated. We investigated the links between direct anthropogenic pressures, socioeconomic settings, and the coverage of vertebrate taxa by China's PA network, and indicated that high socioeconomic status and low levels of human pressure correlate with high species coverage, with threatened mammals more effectively conserved than reptiles or amphibians. Positive links between conservation outcomes and socioeconomic progress appear linked to local livelihood improvements triggering positive perceptions of local PAs-aided further by ecological compensation and tourism schemes introduced in wealthy areas and reinforced by continued positive conservation outcomes. Socioeconomic development of China's less developed regions might assist regional PA efficiency and achievement of the Kunming-Montreal Global Biodiversity Framework, while also addressing potential shortcomings from an insufficient past focus on socioeconomic impacts for biodiversity conservation.

Corneal densitometry: an innovative method to quantitatively evaluate corneal changes after phacovitrectomy.

BACKGROUND: To quantitatively investigate corneal changes and the correlation between corneal densitometry (CD) and endothelial parameters after phacovitrectomy. METHODS: Thirty-eight eyes with idiopathic full-thickness macular holes (iFTMHs) and cataracts underwent phacovitrectomy. Examinations were conducted at baseline and Day 1, Day 7, Month 1, and Month 3 postoperatively. CD and central corneal thickness (CCT) were measured using Pentacam. Corneal endothelial cell density (ECD), coefficient of variation (CV), and hexagonality (HEX) were measured using specular microscopy. RESULTS: ECD and HEX significantly decreased after surgery and the change in HEX occurred prior to CV. CCT increased immediately after surgery and recovered 3 months postoperatively. CD values increased significantly 1 day after surgery and then gradually decreased. For CD in the 0-2 mm zone, it took 1 month to recover in the central and posterior layers and 3 months in the anterior and total layers. For CD in the 2-6 mm zone, the central layer recovered at Day 7, the anterior and total layers recovered at 1 month, and the posterior layer did not recover until 3 months postoperatively. The CD within all layers in the 0-2 mm zone was positively correlated with CCT. Posterior CD in the 0-2 mm zone was negatively correlated with ECD and HEX. CONCLUSIONS: CD is not only correlated with CCT, ECD, and HEX but also reflects the state of the whole cornea and each layer. CD can be an objective, rapid, and noninvasive tool that reflects corneal health and undetectable edema and monitors the process of lesion repair. TRIAL REGISTRATION: This study was registered with the Chinese Clinical Trial Registry (31/10/2021, ChiCTR2100052554).

IGF2BP3-NRF2 axis regulates ferroptosis in hepatocellular carcinoma.

Poor sensitivity to sorafenib has been an important constraint on the efficacy of targeted therapy in advanced hepatocellular carcinoma (HCC). Therefore, it is particularly important to explore effective therapeutic targets to improve the sensitivity of HCC cells to sorafenib. Upregulation of IGF2BP3 is strongly associated with tumor invasion, early recurrence and poor prognosis in various human cancers, including HCC, but its roles in the sorafenib treatment of HCC remain unclear. In our study, IGF2BP3 knock-down significantly promoted ferroptosis in HCC cells through the evaluation of the Reactive Oxygen Species (ROS), Fe2+ and malondialdehyde (MDA) levels after sorafenib administration. In addition, NRF2 mRNA was identified as an important target of IGF2BP3 by bioinformatics analysis, RNA binding protein immunoprecipitation (RIP) and RNA pulldown experiments. More importantly, IGF2BP3, as an m6A (N6-Methyladenosine) reader, was shown to promote the stability of NRF2 mRNA by reading its m6A modification. Similar results were obtained from in vivo experiments. In summary, our study uncovered the role of IGF2BP3-NRF2 axis on ferroptosis in HCC, providing significant evidence for new anti-cancer strategies aimed at improving the efficacy of sorafenib.

Distant metastasis in medullary thyroid carcinoma: Clinical outcomes and implications of T stage.

BACKGROUND: The eighth edition of the American Joint Committee on Cancer tumour, node, and metastasis staging system did not take T stage into consideration when evaluating Stage IV C medullary thyroid carcinoma (MTC) patients. The aim of this study is to investigate the clinical outcomes and implications of T stage in this population. METHODS: Eligible patients from the Surveillance, Epidemiology, and End Results database and the Department of Thyroid Surgery in West China Hospital of Sichuan University and who were diagnosed with Stage IV C MTC were included in this study. The overall survival (OS), the cancer-specific survival (CSS), and the precise cause of MTC-induced death were analysed. The potential risk factors, including the T stage, in the OS and CSS were evaluated by univariate and multivariate Cox regression models. RESULTS: This retrospective study enroled 204 Stage IV C MTC patients. The 5- and 10-year OS rates were 31.8% and 17.1%, respectively, and the 5- and 10-year CSS rates were 40.4% and 22.5%, respectively. More importantly, the rates of MTC-induced death between primary or distant metastatic lesions in Stage IV C MTC patients were comparable in our institution. Additionally, the univariate and multivariate analyses demonstrated that the presence of an advanced T stage was an independent prognostic factor for both the OS (T4 vs. T1-T3, hazard ratio [HR]: 1.714, 95% confidence interval [CI]: 1.175-2.500, p = .005) and the CSS (T4 vs. T1-T3, HR: 1.848, 95% CI: 1.229-2.780, p = .003). CONCLUSION: To achieve a better risk stratification, further classification of Stage IV C MTC patients by the T stage may be preferable.

An optional surgical technique for obtaining lamellar donor grafts: a pilot study.

BACKGROUND: To evaluate the surface quality and thickness uniformity of lamellar donor grafts using an optional surgical technique called reversed manual dissection (RMD) in porcine corneas. METHODS: Twenty-four paired porcine corneas (48 eyes) were numbered 1 to 24 and divided into 6 groups. All left corneas were assigned to conventional manual dissection (CMD), and all right corneas were assigned to RMD. Each group contained 8 corneas. For Groups I, II, and III, 30, 50, and 70% of the entire corneal thickness was dissected using CMD. For groups IV, V, and VI, 70, 50, and 30% of the entire corneal thickness was dissected using RMD. The residual stromal thickness was examined by anterior segment optical coherence tomography (ASOCT) to assess the thickness uniformity and scanning electron microscopy (SEM) to assess the surface quality. RESULTS: The thickness uniformity of the lamellar grafts between each paired group was not significantly different (p > 0.05). The qualitative surface roughness grading (QiSR) evaluated by masked observers through SEM was significantly higher in the RMD groups (p < 0.001). The quantitative surface roughness grading (QnSR) acquired from the Mountains software was significantly lower in the RMD groups (p < 0.001). CONCLUSIONS: RMD is an optional surgical technique for obtaining porcine lamellar grafts. The thickness uniformity of RMD is comparable to that of CMD, and a smoother surface with fewer ridges and roughness is achieved compared to CMD.

Drug Resistance (Dapsone, Rifampicin, Ofloxacin) and Resistance-Related Gene Mutation Features in Leprosy Patients: A Systematic Review and Meta-Analysis.

Dapsone (DDS), Rifampicin (RIF) and Ofloxacin (OFL) are drugs recommended by the World Health Organization (WHO) for the treatment of leprosy. In the context of leprosy, resistance to these drugs occurs mainly due to mutations in the target genes (Folp1, RpoB and GyrA). It is important to monitor antimicrobial resistance in patients with leprosy. Therefore, we performed a meta-analysis of drug resistance in Mycobacterium leprae and the mutational profile of the target genes. In this paper, we limited the study period to May 2022 and searched PubMed, Web of Science (WOS), Scopus, and Embase databases for identified studies. Two independent reviewers extracted the study data. Mutation and drug-resistance rates were estimated in Stata 16.0. The results demonstrated that the drug-resistance rate was 10.18% (95% CI: 7.85-12.51). Subgroup analysis showed the highest resistance rate was in the Western Pacific region (17.05%, 95% CI:1.80 to 13.78), and it was higher after 2009 than before [(11.39%, 7.46-15.33) vs. 6.59% (3.66-9.53)]. We can conclude that the rate among new cases (7.25%, 95% CI: 4.65-9.84) was lower than the relapsed (14.26%, 95 CI%: 9.82-18.71). Mutation rates of Folp1, RpoB and GyrA were 4.40% (95% CI: 3.02-5.77), 3.66% (95% CI: 2.41-4.90) and 1.28% (95% CI: 0.87-1.71) respectively, while the rate for polygenes mutation was 1.73% (0.83-2.63). For further analysis, we used 368 drug-resistant strains as research subjects and found that codons (Ser, Pro, Ala) on RpoB, Folp1 and GyrA are the most common mutation sites in the determining region (DRDR). In addition, the most common substitution patterns of Folp1, RpoB, and GyrA are Pro→Leu, Ser→Leu, and Ala→Val. This study found that a higher proportion of patients has developed resistance to these drugs, and the rate has increased since 2009, which continue to pose a challenge to clinicians. In addition, the amino acid alterations in the sequence of the DRDR regions and the substitution patterns mentioned in the study also provide new ideas for clinical treatment options.

HDAC1-Smad3-mSin3A complex is required for Smad3-induced transcriptional inhibition of hepatocyte growth factor receptor in human lung cancers.

c-Met hyperactivity has been observed in numerous neoplasms. Several researchers have shown that the abnormal activation of c-Met is mainly caused by transcriptional activation. However, the molecular mechanism behind this transcriptional regulation is poorly understood. Here, we suggest that Smad3 negatively regulates the expression and activation of c-Met via a transcriptional mechanism. We explore the molecular mechanisms that underlie Smad3-induced c-Met transcription inhibition. We found in contrast to the high expression of c-Met, Smad3 showed low protein and mRNA levels. Smad3 and c-Met expressions were inconsistent between lung cancer tissues and cell lines. We also found that Smad3 overexpression suppresses whereas Smad3 knockdown significantly promotes Epithelial-Mesenchymal Transition and production of the angiogenic factors VEGF, CTGF and COX-2 through the ERK1/2 pathway. In addition, Smad3 overexpression decreases whereas Smad3 knockdown significantly increases protein and mRNA levels of invasion-related ß-catenin and FAK through the PI3K/Akt pathway. Furthermore, using the chromatin immunoprecipitation analysis method, we demonstrate that a transcriptional regulatory complex consisting of HDAC1, Smad3 and mSin3A binds to the promoter of the c-Met gene. By either silencing endogenous mSin3A expression with siRNA or by pretreating cells with a specific HDAC1 inhibitor (MS-275), Smad3-induced transcriptional suppression of c-Met could be effectively attenuated. These results demonstrate that Smad3-induced inhibition of c-Met transcription depends on of a functional transcriptional regulatory complex that includes Smad3, mSin3A and HDAC1 at the c-Met promoter. Collectively, our findings reveal a new regulatory mechanism of c-Met signaling, and suggest a potential molecular target for the development of anticancer drugs.

Cooperation with partners of differing social experience: An fNIRS-based hyperscanning study.

Previous studies have shown the brain synchronization of all team members while completing a collaborative task. Moreover, this effect is influenced by a team's compositional elements, such as gender (opposite or same) or relationships (i.e., friends, lovers, or strangers) among team members. However, whether interpersonal brain synchronization (IBS) is affected by team members' experience, as well as the temporal dynamics of such brain synchronization, remains to be investigated. In the current study, we combined behavioral methods and functional near-infrared spectroscopy-based hyperscanning to examine the effect of member experience on team cooperation by an adopted continuous joint drawing task with 21 student-student dyads (S-S dyads) and 22 teacher-student dyads (T-S dyads). The results revealed that team members with differing experiences (T-S dyads) perform better than those with similar ones (S-S dyads). Moreover, we observed IBS in the left frontopolar region (channel 11). However, we did not observe significant changes of the task-related IBS across time. Besides, IBS was negatively correlated with the participants' behavioral performance. Our findings demonstrate the importance of social experience in teamwork in the real world and suggest a possible mechanism for cooperation from a temporal and spatial perspective.

Current operation state of wastewater treatment plants in urban China.

Wastewater treatment plants (WWTPs) in urban China generally face operational difficulties. To comprehensively analyze the current operation status and determine the factors that caused difficulties in urban domestic WWTPs, we conducted a questionnaire survey, and 18 operating conditions of scale-level WWTPs in seven regions in China were investigated. The research results showed that, of the 467-urban domestic WWTPs surveyed in China, approximately 63.17% of the WWTPs' hydraulic loading rates (HLRs) are greater than 80%, 67.02% of the WWTPs have mixed liquor suspended solids (MLSS) concentrations greater than 4000 mg/L, and 95.93% of the WWTPs' mixed liquor volatile suspended solids to mixed liquor suspended solids (MLVSS/MLSS) ratios are lower than 0.7. The WWTP energy consumption accounts for approximately 1% of the national electricity consumption, and approximately 100,000 tons of various chemicals are consumed every year. A high HLR, a high activated sludge concentration and low sludge activity, design value mismatches with actual values, high energy and material consumption, etc. have become the main operational difficulties of urban domestic WWTPs. In this study, the main barriers that hinder improving the operating efficiency of WWTPs were investigated, the causes of operational dilemmas were analyzed, and pathways were jointly explored to solve them. This study may provide valuable implications for industry practitioners with a comprehensive grasp of the industry's current status and related policy formulation.

The C-terminal of the α1b-adreneroceptor is a key determinant for its structure integrity and biological functions.

The C-terminal of G protein-coupled receptors is now recognized as being important for G protein activation and signaling function. To detect the role of C-terminal tail in receptor activation, we used the α1b-AR, which has a long C-terminal of 164 amino acids. We constructed the intramolecular FRET sensors, in which the C-terminal was truncated to 10 (∆C-10), 20 (∆C-20), 30 (∆C-30), 50 (∆C-50), 70 (∆C-70), or 90 (∆C-90). The truncated mutants of ∆C-10, ∆C-20, or ∆C-30 cannot induce FRET signal changes and downstream ERK1/2 phosphorylation. However, the truncated mutants of ∆C-50, ∆C-70, or ∆C-90 induce significant FRET signal changes and downstream ERK1/2 phosphorylation, especially ∆C-90. This is particularly true in the case of the ∆C-90, ∆C-70, or ∆C-50 which retained the potential phosphorylation sites (Ser401, Ser404, Ser408, or Ser410). The ∆C-90 showed an increase in agonist-induced FRET signal changes and ERK1/2 phosphorylation in PKC- or endocytosis-dependent and EGFR-, src-, or ß-arrestin2-independent.

Enhancing the activity and thermal stability of a phthalate-degrading hydrolase by random mutagenesis.

Our previous work has reported that EstJ6 was a phthalate-degrading hydrolase. In the study, a random mutant library was constructed by two rounds of error-prone PCR, three mutants (ET1.1, ET2.1, and ET2.2) with enhanced hydrolytic activity against dibutyl phthalate (DBP) were obtained. The best mutant ET2.2, accumulated three amino acid substitutions (Thr91Met, Ala67Val, and Val249Ile) and exhibited 2.8-fold increase enzyme activity and 2.3-fold higher expression level. Meanwhile, compared with EstJ6, ET2.2 showed over 50% improvement in thermostability (at 50 °C for 1 h) and 1.2-fold increase in 50% methanol tolerance. Kinetic parameters analysis revealed that the Km value for ET2.2 decreased by 60% and the kcat/Km value increased by 166%. The molecular docking indicated that the shortening of hydrogen bond between Ser146-OH and DBP-CO, which may led to an increase in enzyme activity and catalytic efficiency, the enhancement of hydrophobicity of hydrophobic pocket was related to the improvement of organic solvents tolerance, and three hydrophobic amino acid substitutions Thr91Met, Ala67Val, and Val249Ile facilitated to improve the thermal stability and organic solvents tolerance. These results confirmed that random mutagenesis was an effective tool for improving enzyme properties and lay a foundation for practical applications of phthalate-degrading hydrolase in biotechnology and industrial fields.

Regulation of anaerobic fermentation for producing short-chain fatty acids from primary sludge in WWTPs by different alkalis.

Carbon source production from primary sludge in wastewater treatment plants (WWTPs) via anaerobic fermentation process has been paid more attention. However, slow hydrolysis rate and low yield of short-chain fatty acids (SCFAs) limited its application. This study aimed at improving the anaerobic fermentation efficiency of primary sludge by alkali regulation (NaOH, Na2CO3 and Ca(OH)2), and revealing the mechanism. Results showed that three kinds of alkalis allowed enhancing hydrolysis and acidification, and reducing methane production in the anaerobic fermentation process of primary sludge. The Na2CO3 regulation contributed to highest yield and productivity of SCFAs, reaching 1626 mg COD/L and 0.189 g COD/g VSS at 4th day, respectively. Microbial community structure analysis indicated that the relative abundance of fermentative microbial community was improved in the alkali regulation system, where methanogenic archaea was effectively inhibited. The continuous flow experiment further verified that the Na2CO3 regulation could steadily increase yield of SCFAs in the anaerobic fermentation process of primary sludge, and the yield was also the highest among three kinds of alkali regulation.

Modeling sulfide production in full flow concrete sewers based on the HRT variation of sewerage.

The corrosion and odor in concrete sewers are mainly related to the sulfide production, which is, under certain circumstances, directly proportional to the hydraulic retention time (HRT) of the sewer. To reduce the corrosion and control the odor in concrete sewers, it is necessary to model the production of sulfide in the concrete sewers with different HRTs. However, previous researches were mostly carried out in simulated Perspex-made sewers, and the obtained theoretical formulas based on the Monod equation were impractical because of the complexity. An actual concrete pipe with domestic sewage was employed in this study to obtain a simple but practical model, which can be applied to quantitively describe the sulfide production according to the HRT of the sewer and the chemical oxygen demand (COD) of the sewage. The empirical equation obtained was rs = (0.045 × lnHRT + 0.071) × ([COD] - b)0.6, the coefficient is a logarithmic function of the HRT, and the sulfide production rate and COD have a power relationship. Based on the data of COD and HRT obtained in the realistic sewer, the production of sulfide in the sewer can be predicted for better maintaining sewers through sulfide control.

Behavioral and brain synchronization differences between expert and novice teachers when collaborating with students.

Differences in behavior and neural mechanisms between expert and novice teachers when collaborating with students are poorly understood. This study investigated whether expert teachers do better in collaborating with students than novice teachers and explored the neural basis of such differences. Novice teacher and student (NT-S) dyads and expert teacher and student (ET-S) dyads were recruited to complete an interactive task consisting of a cooperation and an independent condition. During the experiment, neural activity in the prefrontal cortex of the participants was recorded with functional near-infrared spectroscopy. The results show higher accuracy for the ET-S dyads than the NT-S dyads in the cooperation condition; however, no difference was found in the independent condition. Increased interpersonal brain synchronization (IBS) was detected in the left dorsolateral prefrontal cortex of participants in ET-S dyads, but not in NT-S dyads in the cooperation condition. Moreover, an interaction effect of dyad type and conditions on IBS was observed, revealing IBS was stronger in ET-S dyads than in NT-S dyads. In ET-S dyads, IBS was positively correlated with the teachers' perspective-taking ability and accuracy. These findings suggest that expert teachers collaborate better with students than novice teachers, and IBS might be the neural marker for this difference.

Identification and characterization of a novel phthalate-degrading hydrolase from a soil metagenomic library.

Phthalate esters have raised public concerns owing to their effects on the environment and human health. We identified a novel phthalate-degrading hydrolase, EstJ6, from a metagenomic library using function-driven screening. Phylogenetic analysis indicated that EstJ6 is a member of family IV esterases. EstJ6 hydrolyzed various dialkyl and monoalkyl phthalate esters, and exhibited high hydrolytic activity (128 U/mg) toward dibutyl phthalate at 40 °C and pH 7.5. EstJ6 hydrolyzed not only common phthalate esters with simple side chains but also diethylhexyl phthalate and monoethylhexyl phthalate, which have complex and long side chains. Site-directed mutagenesis indicated that the catalytic triad residues of EstJ6 consists of Ser146, Glu240, and His270. EstJ6 is therefore a promising biodegradation enzyme, and our study illustrates the advantages of a metagenomic approach in identifying enzyme-coding genes for agricultural, food, and biotechnological applications.

Amino-Functionalized Nitrogen-Doped Graphene-Quantum-Dot-Based Nanomaterials with Nitrogen and Amino-Functionalized Group Content Dependence for Highly Efficient Two-Photon Bioimaging.

We fabricated nanomaterials comprising amino-functionalized and nitrogen-doped graphene quantum dots (amino-N-GQDs) and investigated their photostability and intrinsic luminescence in the near-infrared spectrum to determine their suitability as contrast agents in two-photon imaging (TPI). We observed that amino-N-GQDs with a higher amount of bonded nitrogen and amino-functionalized groups (6.2%) exhibited superior two-photon properties to those with a lower amount of such nitrogen and groups (4.9%). These materials were conjugated with polymers containing sulfur (polystyrene sulfonate, PSS) and nitrogen atoms (polyethylenimine, PEI), forming amino-N-GQD-PSS-PEI specimens (amino-N-GQD-polymers). The polymers exhibited a high quantum yield, remarkable stability, and notable two-photon properties and generated no reactive oxygen species, rendering them excellent two-photon contrast agents for bioimaging. An antiepidermal growth factor receptor (AbEGFR) was used for labeling to increase specificity. Two-photon imaging (TPI) of amino-N-GQD (6.2%)-polymer-AbEGFR-treated A431 cancer cells revealed remarkable brightness, intensity, and signal-to-noise ratios for each observation at a two-photon excitation power of 16.9 nJ pixel-1 under 30 scans and a three-dimensional (3D) depth of 105 µm, indicating that amino-N-GQD (6.2%)-polymer-AbEGFR-treated cells can achieve two-photon luminescence with 71 times less power required for two-photon autofluorescence (1322.8 nJ pixel-1 with 500 scans) of similar intensity. This economy can minimize photodamage to cells, rendering amino-N-GQD-polymers suitable for noninvasive 3D bioimaging.