Optimization of Pinocembrin Biosynthesis in Saccharomyces cerevisiae

The flavonoid pinocembrin and its derivatives have gained increasing interest for their benefits on human health. While pinocembrin and its derivatives can be produced in engineered Saccharomyces cerevisiae, yields remain low. Here, we describe novel strategies for improved de novo biosynthesis of pinocembrin from glucose based on overcoming existing limitations in S. cerevisiae. First, we identified cinnamic acid as an inhibitor of pinocembrin synthesis. Second, by screening for more efficient enzymes and optimizing the expression of downstream genes, we reduced cinnamic acid accumulation. Third, we addressed other limiting factors by boosting the availability of the precursor malonyl-CoA, while eliminating the undesired byproduct 2′,4′,6′-trihydroxy dihydrochalcone. After optimizing cultivation conditions, 80 mg/L pinocembrin was obtained in a shake flask, the highest yield reported for S. cerevisiae. Finally, we demonstrated that pinocembrin-producing strains could be further engineered to generate 25 mg/L chrysin, another interesting flavone. The strains generated in this study will facilitate the production of flavonoids through the pinocembrin biosynthetic pathway

Visible-light promoted atom transfer radical addition-elimination (ATRE) reaction for the synthesis of fluoroalkylated alkenes using DMA as electron-donor

Here, we describe a mild, catalyst-free and operationally-simple strategy for the direct fluoroalkylation of olefins driven by the photochemical activity of an electron donor-acceptor (EDA) complex between DMA and fluoroalkyl iodides. The significant advantages of this photochemical transformation are high efficiency, excellent functional group tolerance, and synthetic simplicity, thus providing a facile route for further application in pharmaceuticals and life sciences

A p-Coumaroyl-CoA Biosensor for Dynamic Regulation of Naringenin Biosynthesis in Saccharomyces cerevisiae

In vivo biosensors that can convert metabolite concentrations into measurable output signals are valuable tools for high-throughput screening and dynamic pathway control in the field of metabolic engineering. Here, we present a novel biosensor in Saccharomyces cerevisiae that is responsive to p-coumaroyl-CoA, a central precursor of many flavonoids. The sensor is based on the transcriptional repressor CouR from Rhodopseudomonas palustris and was applied in combination with a previously developed malonyl-CoA biosensor for dual regulation of p-coumaroyl-CoA synthesis within the naringenin production pathway. Using this approach, we obtained a naringenin titer of 47.3 mg/L upon external precursor feeding, representing a 15-fold increase over the nonregulated system

Yeast optimizes metal utilization based on metabolic network and enzyme kinetics

Metal ions are vital to metabolism, as they can act as cofactors on enzymes and thus modulate individual enzymatic reactions. Although many enzymes have been reported to interact with metal ions, the quantitative relationships between metal ions and metabolism are lacking. Here, we reconstructed a genome-scale metabolic model of the yeast Saccharomyces cerevisiae to account for proteome constraints and enzyme cofactors such as metal ions, named CofactorYeast. The model is able to estimate abundances of metal ions binding on enzymes in cells under various conditions, which are comparable to measured metal ion contents in biomass. In addition, the model predicts distinct metabolic flux distributions in response to reduced levels of various metal ions in the medium. Specifically, the model reproduces changes upon iron deficiency in metabolic and gene expression levels, which could be interpreted by optimization principles (i.e., yeast optimizes iron utilization based on metabolic network and enzyme kinetics rather than preferentially targeting iron to specific enzymes or pathways). At last, we show the potential of using the model for understanding cell factories that harbor heterologous iron-containing enzymes to synthesize high-value compounds such as p-coumaric acid. Overall, the model demonstrates the dependence of enzymes on metal ions and links metal ions to metabolism on a genome scale

Total and horizontal distances of the foveal stereotaxic displacement can be prognostic indicators for patients with idiopathic epiretinal membrane

IntroductionThis study aimed to examine the foveal stereo deviations in the different ectopic inner foveal layer (EIFL) stages of idiopathic epiretinal membrane (iERM) and assess its predictive utility for the baseline and postoperative best-corrected visual acuity (BCVA).MethodsBased on the calculational combination of foveal displacements in the horizontal and vertical axial optical coherence tomography (OCT) images, the foveal stereotaxic displacement was estimated through the total distance (TD, the distance from the foveal bottom to the inner edge of displaced central foveal) and horizontal distance (HD, projection of the TD in the retinal plane). The preoperative TD, HD, and other OCT- and OCT angiography (OCTA)-related indicators were obtained. The correlations between structural parameters and baseline and postoperative BCVA were evaluated through correlation and multiple linear regression analyses.ResultsIn patients with advanced EIFL stage, there was a significant increase in the HD, TD, baseline log of the minimum angle of resolution unit for BCVA, central macular thickness (CMT), acircularity index, and incidence of microcystic macular edema (MME; p < 0.05). Further, they showed a decreased foveal avascular zone (FAZ) area and perimeter (p < 0.001). HD, TD, CMT, MME, FAZ area, and FAZ perimeter were significantly correlated with the baseline and postoperative BCVA (p < 0.05). TD had the highest correlation indexic and was an individual predictor of the baseline and postoperative BCVA. Moreover, FD-300 and MME were individual predictors of postoperative BCVA.DiscussionStereoscopic foveal deviations significantly correlated with the baseline and postoperative visual acuity. TD may be used as an independent prognostic factor for BCVA

Quantitative assessment of retinal microvascular remodeling in eyes that underwent idiopathic epiretinal membrane surgery

Purpose: To explore the surgical outcomes of the macular microvasculature and visual function in eyes with idiopathic epiretinal membrane (iERM) using spectral-domain optical coherence tomography angiography (SD-OCTA).Methods: This observational, cross-sectional study included 41 participants who underwent iERM surgery with a 3-month (3M) follow-up. Forty-one healthy eyes formed the control group. The assessments included best-corrected visual acuity (BCVA) and mean sensitivity (MS) by microperimetry and SD-OCTA assessment of vessel tortuosity (VT), vessel density (VD), foveal avascular zone, and retinal thickness (RT).Results: The findings showed statistically significant differences in VT, foveal avascular zone parameters, RT, BCVA, and MS between the iERM and control groups (p < 0.05). After iERM surgery, the macular VT, SCP VD, and RT decreased significantly (p < 0.01) while the DCP VD increased (p = 0.029). The BCVA improved significantly (p < 0.001) and was associated with the MS (rs = −0.377, p = 0.015). MS was associated with the SCP VD and RT at 3M (SCP VD rs = 0.511, p = 0.001; RT rs = 0.456, p = 0.003). In the superior quadrant, the MS improved significantly (p < 0.001) and the improvement of MS was associated with the reduction of VT (β = −0.330, p = 0.034).Conclusion: Microcirculatory remodeling and perfusion recovery were observed within 3 months after iERM surgery. VT was a novel index for evaluating the morphology of the retinal microvasculature in eyes with iERM and was associated with MS in the superior quadrant

Prediction by a multiparametric magnetic resonance imaging-based radiomics signature model of disease-free survival in patients with rectal cancer treated by surgery

ObjectiveThe aim of this study was to assess the ability of a multiparametric magnetic resonance imaging (MRI)-based radiomics signature model to predict disease-free survival (DFS) in patients with rectal cancer treated by surgery.Materials and methodsWe evaluated data of 194 patients with rectal cancer who had undergone radical surgery between April 2016 and September 2021. The mean age of all patients was 62.6 ± 9.7 years (range: 37–86 years). The study endpoint was DFS and 1132 radiomic features were extracted from preoperative MRIs, including contrast-enhanced T1- and T2-weighted imaging and apparent diffusion coefficient values. The study patients were randomly allocated to training (n=97) and validation cohorts (n=97) in a ratio of 5:5. A multivariable Cox regression model was used to generate a radiomics signature (rad score). The associations of rad score with DFS were evaluated using Kaplan–Meier analysis. Three models, namely a radiomics nomogram, radiomics signature, and clinical model, were compared using the Akaike information criterion.ResultThe rad score, which was composed of four MRI features, stratified rectal cancer patients into low- and high-risk groups and was associated with DFS in both the training (p = 0.0026) and validation sets (p = 0.036). Moreover, a radiomics nomogram model that combined rad score and independent clinical risk factors performed better (Harrell concordance index [C-index] =0.77) than a purely radiomics signature (C-index=0.73) or clinical model (C-index=0.70).ConclusionAn MRI radiomics model that incorporates a radiomics signature and clinicopathological factors more accurately predicts DFS than does a clinical model in patients with rectal cancer

Development and validation of a preoperative MRI-based radiomics nomogram to predict progression-free survival in patients with clival chordomas

ObjectivesThe aim of this study was to establish and validate a MRI-based radiomics nomogram to predict progression-free survival (PFS) of clival chordoma.MethodsA total of 174 patients were enrolled in the study (train cohort: 121 cases, test cohort: 53 cases). Radiomic features were extracted from multiparametric MRIs. Intraclass correlation coefficient analysis and a Lasso and Elastic-Net regularized generalized linear model were used for feature selection. Then, a nomogram was established via univariate and multivariate Cox regression analysis in the train cohort. The performance of this nomogram was assessed by area under curve (AUC) and calibration curve.ResultsA total of 3318 radiomic features were extracted from each patient, of which 2563 radiomic features were stable features. After feature selection, seven radiomic features were selected. Cox regression analysis revealed that 2 clinical factors (degree of resection, and presence or absence of primary chordoma) and 4 radiomic features were independent prognostic factors. The AUC of the established nomogram was 0.747, 0.807, and 0.904 for PFS prediction at 1, 3, and 5 years in the train cohort, respectively, compared with 0.582, 0.852, and 0.914 in the test cohort. Calibration and risk score stratified survival curves were satisfactory in the train and test cohort.ConclusionsThe presented nomogram demonstrated a favorable predictive accuracy of PFS, which provided a novel tool to predict prognosis and risk stratification. Our results suggest that radiomic analysis can effectively help neurosurgeons perform individualized evaluations of patients with clival chordomas

Perforating scleral vessels adjacent to myopic choroidal neovascularization achieved a poor outcome after intravitreal anti-VEGF therapy

BackgroundThis study aimed to summarize the features of perforating scleral vessels (PSVs) in patients with myopic choroidal neovascularization (CNV) (mCNV) using optical coherence tomography angiography (OCTA) and to identify the associations with the response after intravitreal anti-vascular endothelial growth factor (anti-VEGF) therapy.MethodsA consecutive series of naïve patients who had mCNV and received intravitreal anti-VEGF therapy with a follow-up duration of 12 months or more were enrolled. The prevalence, location, and branches of PSVs were analyzed. Projection-resolved OCTA (PR-OCTA) was used to analyze the neovascular signals between CNV and PSVs. Best corrected visual acuity (BCVA) and central macular thickness (CMT) were measured. The proportion of CMT change relative to baseline was used to assess therapeutic response.ResultsA total of 44 eyes from 42 patients with mCNV were enrolled. PSVs were identified in 41 out of 44 eyes. Branches were identified in the PSVs of 24 eyes (57.14%), and 20 eyes did not have PSV branches (47.62%). In eight eyes (18.18%), PSVs were adjacent to mCNV, and in 36 eyes (81.82%), PSVs were not adjacent to mCNV. After anti-VEGF therapy for mCNV, BCVA increased (F = 6.119, p < 0.001) and CMT decreased (F = 7.664, p < 0.001). In the eyes where PSVs were adjacent to mCNV, BCVA improvements (F = 7.649, p = 0.009) were poor, and changes in CMT were small.ConclusionThe eyes with PSVs adjacent to mCNV showed poor therapeutic responses after intravitreal anti-VEGF therapy