Water-Free Cyclosporine Ophthalmic Solution vs Vehicle for Dry Eye Disease: A Randomized Clinical Trial.

Importance: Dry eye disease (DED) is a prevalent eye disorder. Cyclosporine is an effective immunomodulator that is widely used in DED; however, due to its highly hydrophobic nature, delivery of cyclosporine to the ocular surface is challenging. Objective: To evaluate the efficacy and safety of SHR8028, a water-free cyclosporine ophthalmic solution, 0.1%, compared with vehicle in Chinese participants with DED. Design, Setting, and Participants: This was a multicenter, double-blind, vehicle-controlled, phase 3 randomized clinical trial conducted from March 4, 2021, to July 22, 2022. Adult participants with moderate to severe DED were recruited from 12 hospitals in China. Study data were analyzed April 2, 2022, for the primary analysis. Interventions: Following a 14-day run-in period with an artificial tear, participants were randomly assigned (1:1) to receive water-free cyclosporine or vehicle (1 eye drop in each eye twice daily). After a 29-day treatment, participants of both groups were given the option to receive water-free cyclosporine for an additional 12 weeks for longer-term safety assessment. Main Outcomes and Measures: The primary end points were changes from baseline in total corneal fluorescein staining (tCFS) using the National Eye Institute scale and in dryness score on a visual analog scale at day 29. Results: A total of 206 participants (mean [SD] age, 47.8 [14.2] years; 185 female [90%]) were enrolled, with 103 each in the cyclosporine group and the vehicle group. At day 29, the cyclosporine group experienced improved tCFS compared with vehicle (change [Δ] = -1.8; 95% CI, -2.7 to -1.0; P < .001), with a tCFS score decrease from baseline of -4.8 in the cyclosporine group and -3.0 in the vehicle group. Dryness score decreased from baseline in both groups (-19.2 vs -15.4; Δ = -3.8; 95% CI, -9.2 to 1.6; P = .17). During the 29-day treatment, treatment-related adverse events were reported in 15 participants (14.6%) in the cyclosporine group and 11 participants (10.7%) in the vehicle group. Conclusions And Relevance: Results demonstrated superiority of a water-free cyclosporine, 0.1%, eye solution over vehicle in improving tCFS score at day 29 in Chinese participants with DED. However, dryness score (VAS) was not improved at day 29. Trial Registration: ClinicalTrials.gov Identifier: NCT05841043.

Impact of Cooperative Innovation on the Technological Innovation Performance of High-Tech Firms: A Dual Moderating Effect Model of Big Data Capabilities and Policy Support.

The mechanism of cooperative innovation (CI) for high-tech firms aims to improve their technological innovation performance. It is the effective integration of the internal and external innovation resources of these firms, along with the simultaneous reduction in the uncertainty of technological innovation and the maintenance of the comparative advantage of the firms in the competition. This study used 322 high-tech firms as our sample, which were located in 33 national innovation demonstration bases identified by the Chinese government. We implemented a multiple linear regression to test the impact of CI conducted by these high-tech firms at the level of their technological innovation performance. In addition, the study further examined the moderating effect of two boundary conditions-big data capabilities and policy support (PS)-on the main hypotheses. Our study found that high-tech firms carrying out CI can effectively improve their technological innovation performance, with big data capabilities and PS significantly enhancing the degree of this influence. The study reveals the intrinsic mechanism of the impact of CI on the technological innovation performance of high-tech firms, which, to a certain extent, expands the application context of CI and enriches the research perspective on the impact of CI on the innovation performance of firms. At the same time, the findings provide insight for how high-tech firms in the digital era can make reasonable use of data empowerment in the process of CI to achieve improved technological innovation performance.

Association of the endothelial nitric oxide synthase (eNOS) 4a/b polymorphism with the risk of incident diabetic retinopathy in patients with type 2 diabetes mellitus: a systematic review and updated meta-analysis.

OBJECTIVE: To conduct a systematic review and updated meta-analysis on the potential association between endothelial nitric oxide synthase (eNOS) 4a/b polymorphism and the risk of developing diabetic retinopathy (DR) in patients with type 2 diabetes mellitus (T2DM) and to identify possible clinical biomarkers for early screening of DR. MATERIALS AND METHODS: A meta-analysis based on case-control or cross-sectional studies was conducted to examine the correlation between eNOS 4a/b polymorphism and DR. Pooled odds ratio (OR) and 95% confidence interval (CI) were used to estimate the association strength. RESULTS: We included 19 studies, covering 7838 subjects. An association was observed in Caucasians (allelic model: OR = 1.273, 95% CI: 1.006-1.610, p = .045; recessive model: OR = 0.575, 95% CI: 0.371-0.892, p = .014; dominant model: OR = 1.268, 95% CI: 1.052-1.528, p = .013; homozygote model: OR = 1.833, 95% CI: 1.176-2.856, p = .007). Moreover, population-based studies have indicated an association between eNOS 4a/b polymorphism and DR susceptibility. CONCLUSIONS: The present study showed that intron 4a allele of eNOS 4a/b is a risk factor for DR in Caucasians with T2DM. Thus, eNOS 4a/b may be used as a biomarker for the early screening and diagnosis of DR in Caucasian T2DM patients.Key messagesEndothelial nitric oxide synthase 4a/b gene polymorphism is not associated with the risk of developing diabetic retinopathy in the overall population, Asians, or Chinese Han patients with type 2 diabetes. However, 4a is a risk factor for the development of diabetic retinopathy in Caucasians.Endothelial nitric oxide synthase 4a/b gene polymorphism is not associated with the type of diabetic retinopathy.

Correlation between Eye Movements and Asthenopia: A Prospective Observational Study.

Purpose: To analyze the correlation between eye movements and asthenopia so as to explore the possibility of using eye-tracking techniques for objective assessment of asthenopia. Methods: This prospective observational study used the computer visual syndrome questionnaire to assess the severity of asthenopia in 93 enrolled college students (age 20−30) who complained about asthenopia. Binocular accommodation and eye movements during the reading task were also examined. The correlations between questionnaire score and accommodation examination results and eye movement parameters were analyzed. Differences in eye movement parameters between the first and last reading paragraphs were compared. The trends in eye movement changes over time were observed. Results: About 81.7% of the subjects suffered from computer visual syndrome. Computer visual syndrome questionnaire total score was positively correlated with positive relative accommodation (p < 0.05). In the first reading paragraph, double vision was positively correlated with unknown saccades (all p < 0.05). Difficulty focusing at close range was positively correlated with total fixation duration, total visit duration, and reading speed (all p < 0.05). Feeling that sight was worsening was positively correlated with regressive saccades (p < 0.05). However, visual impairment symptoms were not significantly correlated with any accommodative function. In a total 20 min reading, significantly reduced eye movement parameters were: total fixation duration, fixation count, total visit duration, visit count, fixation duration mean, and reading speed (all p < 0.01). The eye movement parameters that were significantly increased were: visit duration mean and unknown saccades (all p < 0.001). Conclusion: Eye tracking could be used as an effective assessment for asthenopia. Among the various eye movement parameters, a decrease in fixation duration and counts may be one of the potential indicators related to asthenopia.

Highly efficient degradation of cypermethrin by a co-culture of Rhodococcus sp. JQ-L and Comamonas sp. A-3.

Cypermethrin is an important synthetic pyrethroid pesticide that widely used to control pests in agriculture. However, extensive use has caused its residue and the metabolite 3-phenoxybenzoic acid (3-PBA) to seriously pollute the environments and agricultural products. In this study, a highly efficient cypermethrin-degrading bacterial consortium was acclimated from long-term pyrethroid-contaminated soil. Two strains, designated JQ-L and A-3, were screened from the consortium, and identified as Rhodococcus sp. and Comamonas sp., respectively. Strain JQ-L transformed 100 mg/L of cypermethrin to 3-PBA within 60 h of incubation; however, 3-PBA could not be further degraded by the strain. Strain A-3 utilized 3-PBA as sole carbon for growth, and completely degraded 100 mg/L of 3-PBA within 15 h of incubation. Co-culture of JQ-L and A-3 completely degraded 100 mg/L of cypermethrin within 24 h of incubation. Furthermore, a complete catabolic pathway of cypermethrin and the metabolite 3-PBA by the co-culture was proposed. This study provided a promising strategy for efficient elimination of cypermethrin residue-contaminated environments and agricultural products.

The impact of policy perception on technology transfer from boundary-spanning perspective-empirical evidence from Chinese technological enterprises.

Technology transfer is an essential source of technological innovation for enterprises, which is conducive to the market transformation of patent achievements and the commercial application of new technologies. Building upon social capital theory, all data analyses were performed using SPSS 22.0 and Amos software with the multiple linear regression method. The study explores the mechanism of policy perception to obtain the technical resources needed for enterprise development through boundary-spanning behavior, with a moderating effect of inter-organizational trust and technological potential gap. The study uses survey data from 125 enterprise teams of 42 technology-based enterprises in China. The results show that policy perceived usefulness and usability significantly promote technology transfer performance and boundary-spanning behavior plays a mediating role between them. Speaking of the influencing factors of technology transfer, technological potential gap significantly moderates the relationship between boundary-spanning and technology transfer performance. In contrast, inter-organizational trust positively moderates the relationship between boundary-spanning and technology transfer performance. The research provides theoretical reference and guidance for enterprises on using policy perception better to improve technology transfer performance in the institutional environment. It also helps inspire enterprises to better deal with the cooperative relationship between relevant stakeholders and achieve win-win cooperation.

The impact of entrepreneurs' cognitive flexibility on the business performance of New Ventures: an empirical study based on Chinese New Ventures.

We developed a "thinking-behavior-outcome" logical framework to explore the effect of entrepreneurs' cognitive flexibility on dual innovation and the performance of new ventures, drawing on social cognitive theory, and collected data from a sample of 293 new ventures through a questionnaire to conduct an empirical analysis. We find that entrepreneurs' cognitive flexibility may indirectly affect the performance of new ventures by influencing their dual innovation activities; resource management capabilities positively moderate the relationship between entrepreneurs' cognitive flexibility and dual innovation. In addition, our study demonstrates that dual innovation equilibrium has a stronger effect on the performance of new ventures than single-dimensional innovation activities. Our study highlights the importance of entrepreneurial cognitive flexibility for new ventures to implement innovation strategies and improve performance. The findings not only help to clarify the relationship between entrepreneurs' cognitive flexibility and new venture performance and to enrich social cognitive theory, but they also provide new perspectives on responses to enhance dual innovation capabilities of new ventures under the influence of COVID-19. Supplementary Information: The online version contains supplementary material available at 10.1007/s12144-022-03532-x.

The TetR Family Repressor HpaR Negatively Regulates the Catabolism of 5-Hydroxypicolinic Acid in Alcaligenes faecalis JQ135 by Binding to Two Unique DNA Sequences in the Promoter of Hpa Operon.

5-Hydroxypicolinic acid (5HPA), an important natural pyridine derivative, is microbially degraded in the environment. Previously, a gene cluster, hpa, responsible for 5HPA degradation, was identified in Alcaligenes faecalis JQ135. However, the transcription regulation mechanism of the hpa cluster is still unknown. In this study, the transcription start site and promoter of the hpa operon was identified. Quantitative reverse transcription-PCR and promoter activity analysis indicated that the transcription of the hpa operon was negatively regulated by a TetR family regulator, HpaR, whereas the transcription of hpaR itself was not regulated by HpaR. Electrophoretic mobility shift assay and DNase I footprinting revealed that HpaR bound to two DNA sequences, covering the -35 region and -10 region, respectively, in the promoter region of the hpa operon. Interestingly, the two binding sequences are partially palindromic, with 3 to 4 mismatches and are complementary to each other. 5HPA acted as a ligand of HpaR, preventing HpaR from binding to promoter region and derepressing the transcription of the hpa operon. The study revealed that HpaR binds to two unique complementary sequences of the promoter of the hpa operon to negatively regulate the catabolism of 5HPA. IMPORTANCE This study revealed that the transcription of the hpa operon was negatively regulated by a TetR family regulator, HpaR. The binding of HpaR to the promoter of the hpa operon has the following unique features: (i) HpaR has two independent binding sites in the promoter of the hpa operon, covering -35 region and -10 region, respectively; (ii) the palindrome sequences of the two binding sites are complementary to each other; and (iii) both of the binding sites include a 10-nucleotide partial palindrome sequence with 3 to 4 mismatches. This study provides new insights into the binding features of the TetR family regulator with DNA sequences.

Prediction of occult tumor progression via platelet RNAs in a mouse melanoma model: a potential new platform for early detection of cancer.

BACKGROUND: Cancer screening provides the opportunity to detect cancer early, ideally before symptom onset and metastasis, and offers an increased opportunity for a better prognosis. The ideal biomarkers for cancer screening should discriminate individuals who have not developed invasive cancer yet but are destined to do so from healthy subjects. However, most cancers lack effective screening recommendations. Therefore, further studies on novel screening strategies are urgently required. METHODS: We used a simple suboptimal inoculation melanoma mouse model to obtain 'pre-diagnostic samples' of mice with macroscopic melanomas. High-throughput sequencing and bioinformatic analysis were employed to identify differentially expressed RNAs in platelet signatures of mice injected with a suboptimal number of melanoma cells (eDEGs) compared with mice with macroscopic melanomas and negative controls. Moreover, 36 genes selected from the eDEGs via bioinformatics analysis were verified in a mouse validation cohort via quantitative real-time PCR. LASSO regression was utilized to generate the prediction models with gene expression signatures as the best predictors for occult tumor progression in mice. RESULTS: These RNAs identified from eDEGs of mice injected with a suboptimal number of cancer cells were strongly enriched in pathways related to immune response and regulation. The prediction models generated by 36 gene qPCR verification data showed great diagnostic efficacy and predictive value in our murine validation cohort, and could discriminate mice with occult tumors from control group (area under curve (AUC) of 0.935 (training data) and 0.912 (testing data)) (gene signature including Cd19, Cdkn1a, S100a9, Tap1, and Tnfrsf1b) and also from macroscopic tumor group (AUC of 0.920 (training data) and 0.936 (testing data)) (gene signature including Ccr7, Cd4, Kmt2d, and Ly6e). CONCLUSIONS: Our proof-of-concept study provides evidence for potential clinical relevance of blood platelets as a platform for liquid biopsy-based early detection of cancer.

Genetic Foundations of Direct Ammonia Oxidation (Dirammox) to N2 and MocR-Like Transcriptional Regulator DnfR in Alcaligenes faecalis Strain JQ135.

Ammonia oxidation is an important process in both the natural nitrogen cycle and nitrogen removal from engineered ecosystems. Recently, a new ammonia oxidation pathway termed Dirammox (direct ammonia oxidation, NH3→NH2OH→N2) has been identified in Alcaligenes ammonioxydans. However, whether Dirammox is present in other microbes, as well as its genetic regulation, remains unknown. In this study, it was found that the metabolically versatile bacterium Alcaligenes faecalis strain JQ135 could efficiently convert ammonia into N2 via NH2OH under aerobic conditions. Genetic deletion and complementation results suggest that dnfABC is responsible for the ammonia oxidation to N2 in this strain. Strain JQ135 also employs aerobic denitrification, mainly producing N2O and trace amounts of N2, with nitrite as the sole nitrogen source. Deletion of the nirK and nosZ genes, which are essential for denitrification, did not impair the capability of JQ135 to oxidize ammonia to N2 (i.e., Dirammox is independent of denitrification). Furthermore, it was also demonstrated that pod (which encodes pyruvic oxime dioxygenase) was not involved in Dirammox and that AFA_16745 (which was previously annotated as ammonia monooxygenase and is widespread in heterotrophic bacteria) was not an ammonia monooxygenase. The MocR-family transcriptional regulator DnfR was characterized as an activator of the dnfABC operon with the binding motif 5'-TGGTCTGT-3' in the promoter region. A bioinformatic survey showed that homologs of dnf genes are widely distributed in heterotrophic bacteria. In conclusion, this work demonstrates that, besides A. ammonioxydans, Dirammox occurs in other bacteria and is regulated by the MocR-family transcriptional regulator DnfR. IMPORTANCE Microbial ammonia oxidation is a key and rate-limiting step of the nitrogen cycle. Three previously known ammonia oxidation pathways (i.e., nitrification, anaerobic ammonia oxidation [Anammox], and complete ammonia oxidation [Comammox]) are mediated by autotrophic microbes. However, the genetic foundations of ammonia oxidation by heterotrophic microorganisms have not been investigated in depth. Recently, a previously unknown pathway, termed direct ammonia oxidation to N2 (Dirammox), has been identified in the heterotrophic bacterium Alcaligenes ammonioxydans HO-1. This paper shows that, in the metabolically versatile bacterium Alcaligenes faecalis JQ135, the Dirammox pathway is mediated by dnf genes, which are independent of the denitrification pathway. A bioinformatic survey suggests that homologs of dnf genes are widely distributed in bacteria. These findings enhance the understanding of the molecular mechanisms of heterotrophic ammonia oxidation to N2.

Efficacy of long-term orthokeratology treatment in children with anisometropic myopia.

AIM: To explore the efficacy of the orthokeratology lens for anisometropic myopia progression. METHODS: A retrospective study was performed. Cycloplegic refraction and axial length (AL) were collected from 50 children (10.52±1.72y) who visited Peking University Third Hospital from July 2015 to August 2020. These children's one eyes (Group A) received monocular orthokeratology lenses at first, after different durations (12.20±6.94mo), their contralateral eyes (Group B) developed myopia and receive orthokeratology as well. The data in 1-year of binocular period were recorded. AL growth rate (difference of follow-up and baseline per month) were compared between two groups by paired t test. Interocular differences of AL were compared by Wilcoxon test. RESULTS: During monocular period, the AL growth rate of the Group A (0.008±0.022 mm/mo) was significantly slower than that of the Group B (0.038±0.018 mm/mo; P<0.0001). However, during binocular period, the AL growth rate of the Group A (0.026±0.014 mm/mo) was significantly faster than that of the Group B (0.016±0.015 mm/mo; P<0.0001). The AL difference between both eyes was 0.6 (0.46) mm, then significantly decreased to 0.22 (0.39) mm when started binocular treatment (P<0.0001). However, it was significantly increased to 0.30 (0.32) mm after a year (P<0.0001), but still significantly lower than baseline (P<0.0001). CONCLUSION: The orthokeratology lens is efficient for control the AL elongation of monocular myopia eyes and reduce anisometropia. For the condition that the contralateral eyes develop myopia and receive orthokeratology lens later, there is no efficiency observed on control interocular difference of AL during binocular treatment.

Characterization of a new heterotrophic nitrification bacterium Pseudomonas sp. strain JQ170 and functional identification of nap gene in nitrite production.

Heterotrophic nitrification bacteria play a critical role in nitrogen cycling and pollution removal. However, the underlying nitrification mechanisms are diverse and have rarely been investigated at the genetic level. In this study, the new heterotrophic nitrifier Pseudomonas sp. strain JQ170 was isolated. Strain JQ170 can utilize ammonia (NH4+-N), nitrite (NO2--N), or nitrate (NO3--N) as sole nitrogen sources, preferring NH4+-N. A ratio of 96.4% of 1.0 mM NH4+-N was removed in 24 h. The optimum pH, temperature, and carbon source for NH4+-N removal were pH 7.0, 30 °C, and citrate, at a C/N ratio of 9-18, respectively. During the NH4+-N removal process, only NO2--N but neither hydroxylamine, NO3--N, nor gaseous nitrogen were detected. A random transposon insertion mutagenesis library of strain JQ170 was constructed. Two NO2--N-production deficient mutants were screened and transposon insertion sites were located in nap genes (which encode periplasmic NO3--N reductase Nap). Further gene knockout and complementation of the napA gene confirmed nap as essential for NO2--N production. The following nitrification processes in strain JQ170 is proposed: NH4+-N to NO3--N in the cytoplasm; then NO3--N to NO2--N in the periplasmic space by Nap; finally, NO2--N secreted out of cells. Overall, this paper provides new insight towards understanding heterotrophic nitrification at the genetic level.

Pseudaminobacter soli sp. nov., Isolated from Paddy Soil Contaminated with Heavy Metals.

A Gram stain-negative, aerobic, rod-shaped strain, designated HC19T, was isolated from heavy metals contaminated paddy soil. The 16S rRNA gene-based phylogenetic analysis indicated that strain HC19T belonged to the genus Pseudaminobacter, and shared 97.0% 16S rRNA gene sequence similarity with P. manganicus JH-7T, and less than 97% similarities with other type strains belonging to the genus. The major cellular fatty acids were C19:0 cyclo ω8c (55.0%) and C18: 1ω7c (18.7%). The major quinone was ubiquinone Q-10. The major polar lipids were phosphatidylmonomethylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine and phosphatidylethanolamine. The average nucleotide identity and digital DNA-DNA hybridization values between the genomes of HC19T and P. manganicus JH-7T were 68.0% and 22%, respectively. The G+C content of the genomic DNA was 63.3 mol%. On the basis of phenotypic, chemotaxonomic, and genotypic data, strain HC19T is considered as a novel species in the genus Pseudaminobacter, for which the name Pseudaminobacter. soli sp. nov. is proposed. The type strain is HC19T (= KCTC 82870T = CCTCC AB 2021107T).

Proteiniclasticum sediminis sp. nov., an obligate anaerobic bacterium isolated from anaerobic sludge.

An obligate anaerobic bacterial BAD-10 T was isolated from anaerobic acetochlor-degrading sludge. The strain was Gram-stain negative, curved rod-shaped, non-motile and non-spore-forming. Growth was observed in PYT medium at pH 6.0-9.0 (optimum, pH 7.5), at 25-47 °C (37 °C) and with 0-1.0% NaCl (w/v, 0%). Strain BAD-10 T could degrade acetochlor. The major fermentation products from peptone-yeast (PY) medium were acetate and butyrate. The predominant cellular fatty acids were iso-C15:0 FAME, anteiso-C15:0 FAME and C16:0 FAME. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the strain BAD-10 T showed closest affiliation to Proteiniclasticum ruminis D3RC-2 T, with a sequence similarity of 97.6%. Genome sequencing revealed a genome size of 2,983,986 bp, a G + C content of 51.4 mol% and protein-coding genes of 3,102. The average nucleotide identity and in silico DNA-DNA hybridization values between strain BAD-10 T and Proteiniclasticum ruminis D3RC-2 T were 71.0% and 20.4%, respectively, which were below the standard thresholds for species differentiation. On the basis of phenotypic, physiological and phylogenetic evidence, strain BAD-10 T represents a novel species in the genus Proteiniclasticum, for which the name Proteiniclasticum sediminis sp. nov. is proposed. Strain BAD-10 T (= CCTCC AB 2021091 T = KCTC 25288 T) is the type strain of the proposed novel species.

Cotinine Hydroxylase CotA Initiates Biodegradation of Wastewater Micropollutant Cotinine in Nocardioides sp. Strain JQ2195.

Cotinine is a stable toxic contaminant, produced as a by-product of smoking. It is of emerging concern due to its global distribution in aquatic environments. Microorganisms have the potential to degrade cotinine; however, the genetic mechanisms of this process are unknown. Nocardioides sp. strain JQ2195 is a pure-culture strain that has been reported to degrade cotinine at micropollutant concentrations. This strain utilizes cotinine as its sole carbon and nitrogen source. In this study, a 50-kb gene cluster (designated cot), involved in cotinine degradation, was predicted based on genomic and transcriptomic analyses. A novel three-component cotinine hydroxylase gene (designated cotA1A2A3), which initiated cotinine catabolism, was identified and characterized. CotA from Shinella sp. strain HZN7 was heterologously expressed and purified and was shown to convert cotinine into 6-hydroxycotinine. H218O-labeling and electrospray ionization-mass spectrometry (ESI-MS) analysis confirmed that the hydroxyl group incorporated into 6-hydroxycotinine was derived from water. This study provides new molecular insights into the microbial metabolism of heterocyclic chemical pollutants. IMPORTANCE In the human body, cotinine is the major metabolite of nicotine, and 10 to 15% of generated cotinine is excreted in urine. Cotinine is a structural analogue of nicotine and is much more stable than nicotine. Increased tobacco consumption has led to high environmental concentrations of cotinine, which may have detrimental effects on aquatic ecosystems and human health. Nocardioides sp. strain JQ2195 is a unique cotinine-degrading bacterium. However, the underlying genetic and biochemical foundations of cotinine degradation are still unknown. In this study, a 50-kb gene cluster (designated cot) was identified by genomic and transcriptomic analyses as being involved in the degradation of cotinine. A novel three-component cotinine hydroxylase gene (designated cotA1A2A3) catalyzed cotinine to 6-hydroxy-cotinine. This study provides new molecular insights into the microbial degradation and enzymatic transformation of cotinine.

MicroRNA-124 regulates lens epithelial cell apoptosis by affecting Fas alternative splicing by targeting polypyrimidine tract-binding protein in age-related cataract.

BACKGROUND: Age-related cataract (ARC) is a primary cause of visual blindness worldwide. Lens epithelial cell (LEC) apoptosis, in which Fas plays an essential role, is a vital cytological basis for cataractogenesis. However, the regulatory mechanism of Fas-dependent LEC apoptosis is not well understood. This study aimed to investigate whether MicroRNA (miRNA)-124 can regulate LEC apoptosis by targeting polypyrimidine tract-binding protein (PTB) and thereby affecting Fas alternative splicing in ARC. METHODS: Lens capsule samples from patients with ARC and cornea donors with a transparent lens were collected. HLE-B3 cells were cultured and treated with hydrogen peroxide (H2 O2 ) to establish an apoptosis model in LECs. The expression of miRNA-124, PTB, Fas, and Fas isoforms in tissues and cell lines was assessed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR), western blotting, polyacrylamide gel electrophoresis, and flow cytometry. The miRNA-124 mimic and inhibitor were transfected into HLE-B3 cells, and the effects of the miRNA-124/PTB/Fas pathway in LECs were assessed by analysis of their related targets. RESULTS: High expression of miRNA-124 and membrane Fas (mFas) mRNA and decreased PTB expression were observed in the lens capsule samples. In cells undergoing H2 O2 -induced apoptosis, mFas expression was increased, accompanied by decreased PTB and increased miRNA-124 expression. Subsequently, miRNA-124 upregulation suppressed PTB expression, elevated the mFas level without affecting total Fas expression and promoted apoptosis; miRNA-124 downregulation exerted the opposite effects. CONCLUSION: This study revealed that miRNA-124 promotes LEC apoptosis in ARC by upregulating mFas through targeted inhibition of PTB. Moreover, the identification of the miRNA-124/PTB/Fas pathway provides novel insight into Fas-dependent LEC apoptosis.

Improved Herbicide Resistance of 4-Hydroxyphenylpyruvate Dioxygenase from Sphingobium sp. TPM-19 through Directed Evolution.

4-Hydroxyphenylpyruvate dioxygenase (HPPD) has attracted extensive interest as a promising target for the genetic engineering of herbicide-resistant crops. However, naturally occurring HPPDs are generally very sensitive to HPPD inhibitors. In this study, random mutagenesis was performed to increase the HPPD inhibitors' resistance of Sphingobium sp. HPPD (SpHPPD). Two mutants, Q258M and Y333F, with improved resistance were obtained. Subsequently, a double-mutant (Q258M/Y333F) was generated through combined mutation. Q258M/Y333F exhibited the highest resistance to four HPPD inhibitors [topramezone, mesotrione, tembotrione, and diketonitrile (DKN)]. The enzyme fitness of Q258M/Y333F to topramezone, mesotrione, tembotrione, and DKN was increased by 4.0-, 4.1-, 4.2-, and 3.2-folds, respectively, in comparison with that of the wild-type. Molecular modeling and docking revealed that Q258M mutation leads to the decrease of enzyme-inhibitor-binding strength by breaking the hydrogen bond between the enzyme and the inhibitor, and Y333F mutation changes the conformational balance of the C-terminal helix H11, which hinders the binding of the inhibitor to the enzyme and thus would contribute to improved herbicide resistance. This study helps to further elucidate the structural basis for herbicide resistance and provides better genetic resources for the genetic engineering of herbicide-resistant crops.

Iridium-Catalyzed Cross-Coupling Reactions of Alkenes by Hydrogen Transfer.

A range of Ru-, Rh-, or Pd-catalyzed vinylic C-H/C-H cross-coupling reactions of olefins have been demonstrated to provide 1,3-dienes, using a quantitative amount of metal oxidants. Although transfer hydrogenation and C-H alkenylation are two important areas that evolved independently, we herein report the first iridium-catalyzed cross-coupling reactions of alkenes by integration of directed C(alkenyl)-H alkenylation and transfer hydrogenation to obviate the usage of a metal oxidant, employing a hydrogen acceptor such as inexpensive chloranil.

The role of aryl hydrocarbon receptor in bone remodeling.

Bone remodeling is a persistent process for maintaining skeletal system homeostasis, and it depends on the dynamic equilibrium between bone-forming osteoblasts and bone-resorbing osteoclasts. Aryl hydrocarbon receptor (Ahr), a ligand-activated transcription factor, plays a pivotal role in regulating skeletal system. In order to better understand the role of Ahr in bone remodeling, we focused on bone remodeling characteristic, and the effects of Ahr on bone formation and differentiation, which suggest that Ahr is a critical control factor in the process of bone remodeling. Moreover, we discussed the impacts of Ahr on several signaling pathways related to bone remodeling, hoping to provide a theoretical basis to improve bone remodeling.