Impact of Starch Binding Domain Fusion on Activities and Starch Product Structure of 4-α-Glucanotransferase.

A broad range of enzymes are used to modify starch for various applications. Here, a thermophilic 4-α-glucanotransferase from Thermoproteus uzoniensis (TuαGT) is engineered by N-terminal fusion of the starch binding domains (SBDs) of carbohydrate binding module family 20 (CBM20) to enhance its affinity for granular starch. The SBDs are N-terminal tandem domains (SBDSt1 and SBDSt2) from Solanum tuberosum disproportionating enzyme 2 (StDPE2) and the C-terminal domain (SBDGA) of glucoamylase from Aspergillus niger (AnGA). In silico analysis of CBM20s revealed that SBDGA and copies one and two of GH77 DPE2s belong to well separated clusters in the evolutionary tree; the second copies being more closely related to non-CAZyme CBM20s. The activity of SBD-TuαGT fusions increased 1.2-2.4-fold on amylose and decreased 3-9 fold on maltotriose compared with TuαGT. The fusions showed similar disproportionation activity on gelatinised normal maize starch (NMS). Notably, hydrolytic activity was 1.3-1.7-fold elevated for the fusions leading to a reduced molecule weight and higher α-1,6/α-1,4-linkage ratio of the modified starch. Notably, SBDGA-TuαGT and-SBDSt2-TuαGT showed Kd of 0.7 and 1.5 mg/mL for waxy maize starch (WMS) granules, whereas TuαGT and SBDSt1-TuαGT had 3-5-fold lower affinity. SBDSt2 contributed more than SBDSt1 to activity, substrate binding, and the stability of TuαGT fusions.

Evaluating cost and benefit of air pollution control policies in China: A systematic review.

China has put great efforts into air pollution control over the past years and recently committed to its most ambitious climate target. Cost and benefit analysis has been widely used to evaluate the control policies in terms of past performance, future reduction potential, and direct and indirect impacts. To understand the cost and benefit analysis for air pollution control in China, we conducted a bibliometric review of more than 100 studies published over the past two decades, including the current research progress, most commonly adopted methods, and core findings. The control target in cost and benefit analysis has shifted in three stages, from individual and primary pollution control, moving to joint prevention of multiple and secondary pollutants, and then towards synergistic control of air pollution and carbon. With the expansion of the research scope, the integrated assessment model has gradually demonstrated the necessity for long-term ex-anti policy simulation, especially for dealing with complex factors. To ensure long-term air quality, climate, public health, and sustainable economic development, substantial evidence from published studies has suggested that China needs to continue its efforts in the upstream adjustment of the energy system and industrial structure with multi-regional and -sector collaboration. This cost and benefit review paper provides decision-makers with the fundamental information and knowledge gaps in air pollution control strategies in China, and direction for facing future challenges.

Intercomparison of equivalent black carbon (eBC) and elemental carbon (EC) concentrations with three-year continuous measurement in Beijing, China.

Due to the lack of black carbon (BC) measurement data in some cases, elemental carbon (EC) is often used as a surrogate of BC, with a simple assumption that they are interchangeable. Such assumption will inevitably lead to uncertainties in radiative forcing estimation and health impact assessment. In order to quantitatively and systematically evaluate the relationship between BC and EC as well as factors responsible for their difference, 3-year collocated equivalent BC (eBC) and EC measurements with 1-h resolution were performed in Beijing, China continuously from 2016 to 2019. EBC concentration was measured by the multi-wavelength aethalometer (AE-33) based on optical analysis, while EC concentration was determined by semi-continuous OC/EC analyzer with thermal-optical method. The results showed that around 90% of eBC concentration was higher than that of EC, with average difference between eBC and EC as 1.21 µg m-3 (accounting for 33% of average eBC in Beijing). EBC and EC concentrations exhibited strong correlation (r = 0.90) during the whole study period, but the slopes (or eBC/EC ratio) and correlation coefficients varied across seasons (spring: 1.67 and 0.94; summer: 0.91 and 0.65; fall: 1.15 and 0.88; winter: 1.09 and 0.91, respectively). Based on the information from shell/core ratios by Single Particle Soot Photometer (SP2), source apportionment results by positive matrix factorization model, and chemical composition of PM2.5, the differences between eBC and EC concentrations were found to be primarily related to BC aging process and secondary components as evidenced by strong positive correlation with secondary species (e.g., secondary organic carbon and nitrate). This study provided seasonal specific conversion factors of eBC and EC in Beijing and helpful reference for other areas, which will contribute new knowledge of carbonaceous aerosol and reduce uncertainty in assessing future climate change and health studies of BC.

The Critical Role of Policy Enforcement in Achieving Health, Air Quality, and Climate Benefits from India's Clean Electricity Transition.

The coal-dominated electricity system poses major challenges for India to tackle air pollution and climate change. Although the government has issued a series of clean air policies and low-carbon energy targets, a key barrier remains enforcement. Here, we quantify the importance of policy implementation in India's electricity sector using an integrated assessment method based on emissions scenarios, air quality simulations, and health impact assessments. We find that limited enforcement of air pollution control policies leads to worse future air quality and health damages (e.g., 14 200 to 59 000 more PM2.5-related deaths in 2040) than when energy policies are not fully enforced (5900 to 8700 more PM2.5-related deaths in 2040), since coal power plants with end-of-pipe controls already emit little air pollution. However, substantially more carbon dioxide will be emitted if low-carbon and clean coal policies are not successfully implemented (e.g., 400 to 800 million tons more CO2 in 2040). Thus, our results underscore the important role of effectively implementing existing air pollution and energy policy to simultaneously achieve air pollution, health, and carbon mitigation goals in India.

Long noncoding RNA MALAT1 participates in the pathological angiogenesis of diabetic retinopathy in an oxygen-induced retinopathy mouse model by sponging miR-203a-3p.

Diabetic retinopathy (DR) is a devastating complication of diabetes. The aim of the present study is to investigate the exact role and mechanism of long noncoding RNA MALAT1 (MALAT1) in the progress of DR. An oxygen-induced retinopathy (OIR) mouse model and high glucose (HG) stimulated human retinal microvascular endothelial cells (HRMECs) were employed to mimic the pathological statues of DR. Quantitative real-time PCR (qRT-PCR) and Western blot results showed that MALAT1, VEGFA, and HIF-1α levels were increased in DR retinal tissues and HG-stimulated HRMECs, whereas the expression of miR-203a-3p was decreased. Knockdown of MALAT1 or upregulation of miR-203a-3p both suppressed HG-induced proliferation, migration, and tube formation of HRMECs. A dual-luciferase reporter assay showed that miR-203a-3p could bind to the predicted seed regions of MALAT1 as evidenced by the reduced luciferase activity. Furthermore, enforced downregulation of miR-203a-3p abolished the suppressive effect of MALAT1 silencing on HRMEC cell migration and tube formation. In conclusion, these data demonstrated that MALAT1 may affect angiogenesis by sponging miR-203a-3p in DR, suggesting that MALAT1 may act as a novel therapeutic target for the treatment of DR.

MiR-203a-3p inhibits retinal angiogenesis and alleviates proliferative diabetic retinopathy in oxygen-induced retinopathy (OIR) rat model via targeting VEGFA and HIF-1α.

Proliferative diabetic retinopathy (PDR) is a common complication of diabetes mellitus, characterized by abnormal retinal angiogenesis. MicroRNA-203-3p (miR-203-3p) was found to be down-regulated in a murine model of proliferative retinopathy. This study was performed to explore the role of miR-203a-3p in retinal angiogenesis of PDR. Firstly, a rat OIR model, which was used to mimic PDR, was established and the OIR rats were treated with scrambled control or miR-203a-3p agomir by intravitreal injection. The results showed that the level of miR-203a-3p was decreased in OIR rats, and forced over-expression of miR-203a-3p inhibited OIR-induced retinal angiogenesis as evidenced by reduced blood vessel profiles and CD31 expression. OIR-induced up-regulation of VEGFA, HIF-α, PCNA, and MMPs in the retina was also counteracted by miR-203a-3p. Additionally, high glucose (HG)-induced proliferation, migration and tube formation of human retinal microvascular endothelial cells (HRMECs) were also dampened by the up-regulation of miR-203a-3p. Dual-luciferase reporter assay showed that miR-203a-3p could specifically bind to the 3'UTR of VEGFA and HIF-1α. Over-expression of VEGFA or HIF-1α restored the tube formation activity of HRMECs suppressed by miR-203a-3p. In conclusion, our findings demonstrate that up-regulation of miR-203a-3p might inhibit pathological retinal angiogenesis of PDR by targeting VEGFA and HIF-1α.

Preloaded vs manually loaded IOL delivery systems in cataract surgery in the largest ambulatory surgery center of northwestern China: an efficiency analysis.

BACKGROUND: To compare the efficiency of preloaded vs manually loaded IOL (P-IOL vs M-IOL, respectively) delivery systems in cataract surgery in the largest ambulatory surgery center of Northwestern China. METHODS: A total of 200 cases were included in this prospective, observational study. Time and motion data were collected in a one- or two-operating room (1-OR or 2-OR, respectively) scenario. A model of the efficiency and revenue implications of introducing a preloaded delivery system for IOLs in cataract surgery was used to estimate the changes in cataract throughput and hospital revenue through transitioning from the M-IOL delivery system to the P-IOL system. RESULTS: In the 1-OR scenario, the mean total case time was 16.9 min using P-IOL, which was a 7.7% reduction compared with M-IOL (P < 0.01). In the 2-OR scenario, the mean total surgeon time was 10.8 min using P-IOL, which was a 7.8% reduction compared with M-IOL (P < 0.05). By switching from M-IOL to P-IOL, annual throughput will increase by 5.2% (960 cases) in the 1-OR scenario and 7.7% (1440 cases) in the 2-OR scenario, accompany by an increase in revenue of approx. 284,352 USD in the 1-OR scenario and approx. 426,528 USD in the 2-OR scenario. CONCLUSION: This report is the first of a comparison of two IOL delivery systems in China using different settings in the scenario. IOL delivery with preloaded systems is time saving in both the 1-OR scenario and the 2-OR scenario. Moreover, switching from the M-IOL delivery system to the P-IOL system holds potential to increase cataract throughput and hospital revenue.

Detecting stochastic multiresonance in neural networks via statistical complexity measure.

This paper employs statistical complexity measure (SCM) to investigate the occurrence of stochastic multiresonance (SMR) induced by noise and time delay in small-world neural networks coupled with FitzHugh-Nagumo (FHN) neurons. Our findings reveal that SCM exhibits four local maxima at four optimal noise levels, providing evidence for the occurrence of quadruple stochastic resonances. When time delay τ is taken into account in the information transmission, under moderate noise levels, SCM shows several local maxima when τ = n T e with n being a positive integer and T e being the period of subthreshold signal. This indicates the appearance of delay-induced SMR at the multiples of the period of subthreshold signal. Intriguingly, at low noise levels, a strong coherence between time delay and neuronal firing dynamics emerges at τ = n T e - 2 , as confirmed by a series of SCM maxima at these time delays. Furthermore, the study demonstrates that by adjusting the degrees and sizes of small-world networks, as well as the coupling strength, it is possible to optimize the strength of delay-induced SMR, thus maximizing the detection capability of subthreshold signal. The research results may provide us with an effective approach for understanding the role of time delay in signal detection and information transmission.

Simvastatin induces caspase-dependent apoptosis and activates P53 in OCM-1 cells.

Simvastatin is a cholesterol-lowering drug which exhibits numerous pleiotropic effects including anti-cancer activity. Yet, the anti-cancer effects in choroidal melanoma remain poorly characterized. Therefore, in this study, we investigated the effects of simvastatin on OCM-1 cells growth, apoptosis and cycle. Simvastatin showed an inhibitory effects on OCM-1 cells viability in dose-dependent (2-10 µM) and time-dependent (24-72 h) manner. Further study suggested that simvastatin-induced inhibition OCM-1 cells proliferation was associated with G1 phase arrest, decreased protein and mRNA expression of proliferation marker cyclin D1, cyclin E, cyclin dependent kinase (CDK)2 and increased expression of CDK inhibitory protein P21. In addition, simvastatin resulted in an increase in levels of reactive oxygen species (ROS) in OCM-1 cells and simvastatin significantly triggered apoptosis in OCM-1 cells, which was characterized by increased chromatin condensation, activation of caspase-9 and cleaved-caspase-3, increased expression mitochondrion-related apoptosis protein of P53, Bax and decreased expression of Bcl2 and iASPP. Collectively, our study demonstrated that simvastatin can efficiently inhibit proliferation and induce apoptosis in OCM-1 cells.

A practical approach to producing the single-arm linear dextrin, a chimeric glucosaccharide containing an (α-1 → 4) linked portion at the nonreducing end of an (α-1 → 6) glucochain.

How to improve the solubility of linear dextrins (LD) and retain their characteristic helix amphiphilic cavities with flexible embedding capability, is a question worth exploring without adding new chemical groups. The strategy presented in this study is to attach a highly flexible (α-1 â†’ 6) glucochain at the reducing end of LD by preparing a new type of dextrin, referred to as single-arm linear dextrin (SLD). In the actual synthesis, an (α-1 â†’ 6) linked oligosaccharide of DP¯ 10.7 (PDI = 1.28) was formed by extension of glucose units onto sucrose (2 M) by using L940W mutant of the glucansucrase GTF180-ΔN firstly. Next using γ-CD as glucosylation donor γ-CGTase extended this (α-1 â†’ 6) glucochain with (α-1 â†’ 4) bonds. SLD is a chimeric glucosaccharide comprising an (α-1 â†’ 4) linked part (DP¯ 10.5) attached to the nonreducing end of an (α-1 â†’ 6) glucochain as verified by enzyme fingerprinting and 1H NMR. Furthermore, SLD was validated to show greatly improved solubility and dispersibility of resveratrol in water, as indicated by a 3.12-fold enhancement over the solubility in the presence of 0.014 M SLD. This study provided a new strategy for solving the solubility problem of LD and opens possibilities for new design of the fine structure of starch-like materials.