The influence of the topographic location of geographic atrophy on vision-related quality of life in nonexudative age-related macular degeneration.

PURPOSE: To examine associations between the topographic distribution of geographic atrophy (GA) and vision-related quality of life (VRQoL). METHODS: This study included 237 eyes from 161 participants in the Age-Related Eye Disease Study (AREDS). GA lesions were manually delineated with color fundus photographs obtained by the AREDS Research Group and atrophic area was measured in an Early Treatment Diabetic Retinopathy Study (ETDRS) grid. VRQoL was measured using the National Eye Institute Visual Function Questionnaire (NEI-VFQ). Area of atrophy in the ETDRS grid subfields was correlated with VRQoL by linear regression modeling. RESULTS: The average area of atrophy in the better and worse eye was 3.43mm2 and 7.15mm2 respectively. In multivariable analysis, VRQoL was not associated with total area of atrophy in the better eye (ß, - 0.53; 95% confidence interval [CI], - 1.11 to 0.05; P = 0.07) or worse eye (ß, 0.12; 95% CI, - 0.32 to 0.55; P = 0.59). However, area of atrophy in the central 1-mm-diameter zone of the better eye was significantly associated with VRQoL when the ETDRS subfields were examined individually (ß, - 14.57; 95% CI, - 27.12 to - 2.02; P = 0.023), grouped into quadrants (ß, - 18.35; 95% CI, - 30.03 to - 6.67; P = 0.002), inner and outer zones (ß, - 17.26; 95% CI, - 29.38 to - 5.14; P = 0.006), or vertical and horizontal zones (ß, - 18.97; 95% CI, - 30.18 to - 7.77; P = 0.001). CONCLUSION: In patients with GA, greater area of atrophy in the central 1-mm-diameter zone of the better eye was independently associated with lower VRQoL, while total area of atrophy in the better or worse eye was not.

Sample Preparation and Imaging Conditions Affect mEos3.2 Photophysics in Fission Yeast Cells.

Photoconvertible fluorescent proteins (PCFPs) are widely used in super-resolution microscopy and studies of cellular dynamics. However, our understanding of their photophysics is still limited, hampering their quantitative application. For example, we do not know the optimal sample preparation methods or imaging conditions to count protein molecules fused to PCFPs by single-molecule localization microscopy in live and fixed cells. We also do not know how the behavior of PCFPs in live cells compares with fixed cells. Therefore, we investigated how formaldehyde fixation influences the photophysical properties of the popular green-to-red PCFP mEos3.2 in fission yeast cells under a wide range of imaging conditions. We estimated photophysical parameters by fitting a three-state model of photoconversion and photobleaching to the time course of fluorescence signal per yeast cell expressing mEos3.2. We discovered that formaldehyde fixation makes the fluorescence signal, photoconversion rate, and photobleaching rate of mEos3.2 sensitive to the buffer conditions likely by permeabilizing the yeast cell membrane. Under some imaging conditions, the time-integrated mEos3.2 signal per yeast cell is similar in live cells and fixed cells imaged in buffer at pH 8.5 with 1 mM DTT, indicating that light chemical fixation does not destroy mEos3.2 molecules. We also discovered that 405-nm irradiation drove some red-state mEos3.2 molecules to enter an intermediate dark state, which can be converted back to the red fluorescent state by 561-nm illumination. Our findings provide a guide to quantitatively compare conditions for imaging mEos3.2-tagged molecules in yeast cells. Our imaging assay and mathematical model are easy to implement and provide a simple quantitative approach to measure the time-integrated signal and the photoconversion and photobleaching rates of fluorescent proteins in cells.

Geographic atrophy severity and mortality in age-related macular degeneration.

PURPOSE: To examine the association between geographic atrophy (GA) disease characteristics and mortality risk. METHODS: We manually delineated color fundus photographs of 209 Age-Related Eye Disease Study (AREDS) participants with GA secondary to age-related macular degeneration to identify total area of atrophy, GA effective radius growth rate, disease laterality, and the presence of foveal center involvement. Associations between GA characteristics and mortality were assessed with Cox proportional hazards models adjusted for health status indicators. RESULTS: During a median follow-up of 6.8 years, 48 (23.0%) participants with GA died. In adjusted models, accounting for age, sex, and health status, participants with total GA area in the highest quartile had a significantly increased risk of all-cause mortality compared to those with total GA area in the lowest quartile (hazard ratio [HR], 3.42; 95% confidence interval [CI], 1.32-8.86; P = 0.011). GA effective radius growth rate, bilateral disease, and the presence of foveal center involvement were not significantly associated with mortality. In a multivariable model, including health status indicators and all GA characteristics, total area of atrophy in the highest quartile remained significantly associated with mortality (HR, 4.65; 95% CI, 1.29-16.70; P = 0.019). CONCLUSIONS: More extensive GA, as indicated by a greater total area of atrophy, was associated with an increased risk of all-cause mortality in our cohort. The extent of GA may reflect the extent of underlying disease processes that contribute to greater mortality risk, further suggesting that GA may be part of a systemic rather than purely ocular disease process.

The roles of cadmium on growth of seedlings by analysing the composition of metabolites in pumpkin tissues.

Changes in the types and contents of metabolites in plants can occur in response to environmental stress. In this study, pumpkin seeds were cultivated in a cadmium ion solution (cadmium sulfate) for 7 days, and growth parameters, antioxidant enzyme activities, and metabolites in the root, stem, and leaf were analyzed. The results showed that cadmium accumulation characteristics were in the order of root > stem > leaf. Cadmium restrained root growth and promoted superoxide dismutase, peroxidase, catalase activities in the root, but inhibited their activities in the leaf. Cadmium did not change the total biomass of pumpkin seedlings. Orthogonal partial least squares (OPLS) analyses were conducted to detect the relationships between fresh weight and metabolites. These analyses revealed that maltose had significantly positive relationships with the fresh weight of the root, stem, and leaf. Cadmium influenced glyoxylate and dicarboxylate metabolism, aminoacyl-tRNA biosynthesis, sulfur metabolism, butanoate metabolism, alanine, aspartate and glutamate metabolism, glutathione metabolism, glycine, serine and threonine metabolism in the root; glycolysis/gluconeogenesis in the stem; and biosynthesis of unsaturated fatty acids, galactose metabolism, cutin, suberine and wax biosynthesis in the leaf. It is important that cadmium inhibited root growth by inhibiting carbohydrate transport from the leaf to the root and promoted leaf growth by the accumulation of carbohydrates in the leaf. Furthermore, cadmium also restrained amino acid metabolism in the root of pumpkin seedlings. These results provide new information about how pumpkin seedlings respond to cadmium stress.

Reactive oxygen species-mediated BIN2 activity revealed by single-molecule analysis.

Much evidence has shown that reactive oxygen species (ROS) regulate several plant hormone signaling cascades, but little is known about the real-time kinetics and the underlying molecular mechanisms of the target proteins in the brassinosteroid (BR) signaling pathway. In this study, we used single-molecule techniques to investigate the true signaling timescales of the major BR signaling components BRI1-EMS-SUPPRESSOR 1 (BES1) and BRASSINOSTEROID INSENSITIVE 2 (BIN2) of Arabidopsis thaliana. The rate constants of BIN2 associating with ATP and phosphorylating BES1 were determined to be 0.7 ± 0.4 mM-1  s-1 and 2.3 ± 1.4 s-1 , respectively. Interestingly, we found that the interaction of BIN2 and BES1 was oxygen-dependent, and oxygen can directly modify BIN2. The activity of BIN2 was switched on via modification of specific cysteine (Cys) residues, including C59, C95, C99 and C162. The mutation of these Cys residues inhibited the BR signaling outputs. These findings demonstrate the power of using single-molecule techniques to study the dynamic interactions of signaling components, which is difficult to be discovered by conventional physiological and biochemical methods.

Natural History of Autosomal Recessive Stargardt Disease in Untreated Eyes: A Systematic Review and Meta-analysis of Study- and Individual-Level Data.

TOPIC: Systematic review and meta-analysis of the natural history of autosomal recessive Stargardt disease (STGD1). CLINICAL RELEVANCE: Controversy exists regarding the progression pattern of atrophic lesions secondary to STGD1, and the reported growth rates vary widely among studies. METHODS: We searched in 6 literature databases up through March 15, 2019, to identify studies that monitored atrophy progression by fundus autofluorescence in untreated eyes with STGD1 for 6 months or more. We analyzed both study- and individual-level data from the included studies using 3 models: the area linear model (ALM), radius linear model (RLM), and area exponential model (AEM), in which the area, radius, and natural log-transformed area changes linearly with time, respectively. A horizontal translation factor was added to each dataset to correct for different participants' entry times into the studies. The best model was determined by the predicted age of lesion onset and dependence of growth rates on baseline lesion sizes. The risk of bias was assessed using the Newcastle-Ottawa scale. RESULTS: Of 3158 articles screened, 7 studies (564 eyes) met our inclusion criteria. Cumulative study- and individual-level datasets fit along a straight line in the RLM after introducing horizontal translation factors to correct for different entry times (r2 = 0.99 and r2 = 0.93, respectively). The growth rate of effective lesion radius was 0.104 mm/year (95% confidence interval, 0.086-0.123 mm/year). The age of atrophy onset predicted by the RLM (22.7±5.0 years) was comparable to the reported age at onset of symptoms (22.1±3.1 years); in contrast, the predictions by the ALM and AEM deviated from this number by more than 5 years. Based on the individual-level data, the effective radius growth rate was independent of the baseline lesion size (r = 0.06); in comparison, the growth rates of area and natural log-transformed area were significantly dependent on the baseline lesion size (r = 0.47 and r = -0.33, respectively). CONCLUSIONS: The progression of STGD1 lesions followed the RLM in both study- and individual-level data. The effective radius growth rate of atrophic lesions could serve as a reliable outcome measure to monitor STGD1 progression.

Dispersive liquid-liquid microextraction based on amine-functionalized Fe3O4 nanoparticles for the determination of phenolic acids in vegetable oils by high-performance liquid chromatography with UV detection.

A novel dispersive liquid-liquid microextraction method based on amine-functionalized Fe3O4 magnetic nanoparticles was developed for the determination of six phenolic acids in vegetable oils by high-performance liquid chromatography. Amine-functionalized Fe3O4 was synthesized by a one-pot solvothermal reaction between Fe3O4 and 1,6-hexanediamine and characterized by transmission electron microscopy and Fourier transform infrared spectrophotometry. A trace amount of phosphate buffer solution (extractant) was adsorbed on bare Fe3O4-NH2 nanoparticles by hydrophilic interaction to form the "magnetic extractant". Rapid extraction could be achieved while the "magnetic extractant" on amine-functionalized Fe3O4 nanoparticles was dispersed in the sample solution by vortexing. After extraction, the "magnetic extractant" was collected by application of an external magnet. Some important parameters, such as pH and volume of extraction and desorption solvents, the extraction and desorption time needed were carefully investigated and optimized to achieve the best extraction efficiency. Under the optimal conditions, satisfactory extraction recoveries were obtained for the six phenolic acids in the range of 84.2-106.3%. Relative standard deviations for intra- and inter-day precisions were less than 6.3 and 10.0%, respectively. Finally, the established method was successfully applied for the determination of six phenolic acids in eight kinds of vegetable oils.

A Fast Transient Response Capacitor-Less LDO with Transient Enhancement Technology.

This paper proposes a fast transient load response capacitor-less low-dropout regulator (CL-LDO) for digital analog hybrid circuits in the 180 nm process, capable of converting input voltages from 1.2 V to 1.8 V into an output voltage of 1 V. The design incorporates a rail-to-rail input and push-pull output (RIPO) amplifier to enhance the gain while satisfying the requirement for low power consumption. A super source follower buffer (SSFB) with internal stability is introduced to ensure loop stability. The proposed structure ensures the steady-state performance of the LDO without an on-chip capacitor. The auxiliary circuit, or transient enhancement circuit, does not compromise the steady-state stability and effectively enhances the transient performance during sudden load current steps. The proposed LDO consumes a quiescent current of 47 µA and achieves 25 µV/mA load regulation with a load current ranging from 0 to 20 mA. The simulation results demonstrate that a settling time of 0.2 µs is achieved for load steps ranging from 0 mA to 20 mA, while a settling time of 0.5 µs is attained for load steps ranging from 20 mA to 0 mA, with an edge time of 0.1 µs.

Automated localization of mandibular landmarks in the construction of mandibular median sagittal plane.

OBJECTIVE: To use deep learning to segment the mandible and identify three-dimensional (3D) anatomical landmarks from cone-beam computed tomography (CBCT) images, the planes constructed from the mandibular midline landmarks were compared and analyzed to find the best mandibular midsagittal plane (MMSP). METHODS: A total of 400 participants were randomly divided into a training group (n = 360) and a validation group (n = 40). Normal individuals were used as the test group (n = 50). The PointRend deep learning mechanism segmented the mandible from CBCT images and accurately identified 27 anatomic landmarks via PoseNet. 3D coordinates of 5 central landmarks and 2 pairs of side landmarks were obtained for the test group. Every 35 combinations of 3 midline landmarks were screened using the template mapping technique. The asymmetry index (AI) was calculated for each of the 35 mirror planes. The template mapping technique plane was used as the reference plane; the top four planes with the smallest AIs were compared through distance, volume difference, and similarity index to find the plane with the fewest errors. RESULTS: The mandible was segmented automatically in 10 ± 1.5 s with a 0.98 Dice similarity coefficient. The mean landmark localization error for the 27 landmarks was 1.04 ± 0.28 mm. MMSP should use the plane made by B (supramentale), Gn (gnathion), and F (mandibular foramen). The average AI grade was 1.6 (min-max: 0.59-3.61). There was no significant difference in distance or volume (P > 0.05); however, the similarity index was significantly different (P < 0.01). CONCLUSION: Deep learning can automatically segment the mandible, identify anatomic landmarks, and address medicinal demands in people without mandibular deformities. The most accurate MMSP was the B-Gn-F plane.

Progress of stimulus responsive nanosystems for targeting treatment of bacterial infectious diseases.

In recent decades, due to insufficient concentration at the lesion site, low bioavailability and increasingly serious resistance, antibiotics have become less and less dominant in the treatment of bacterial infectious diseases. It promotes the development of efficient drug delivery systems, and is expected to achieve high absorption, targeted drug release and satisfactory therapy effects. A variety of endogenous stimulation-responsive nanosystems have been constructed by using special infection microenvironments (pH, enzymes, temperature, etc.). In this review, we firstly provide an extensive review of the current research progress in antibiotic treatment dilemmas and drug delivery systems. Then, the mechanism of microenvironment characteristics of bacterial infected lesions was elucidated to provide a strong theoretical basis for bacteria-targeting nanosystems design. In particular, the discussion focuses on the design principles of single-stimulus and dual-stimulus responsive nanosystems, as well as the use of endogenous stimulus-responsive nanosystems to deliver antimicrobial agents to target locations for combating bacterial infectious diseases. Finally, the challenges and prospects of endogenous stimulus-responsive nanosystems were summarized.

METformin for the MINimization of Geographic Atrophy Progression (METforMIN): A Randomized Trial.

Purpose: Metformin use has been associated with a decreased risk of age-related macular degeneration (AMD) progression in observational studies. We aimed to evaluate the efficacy of oral metformin for slowing geographic atrophy (GA) progression. Design: Parallel-group, multicenter, randomized phase II clinical trial. Participants: Participants aged ≥ 55 years without diabetes who had GA from atrophic AMD in ≥ 1 eye. Methods: We enrolled participants across 12 clinical centers and randomized participants in a 1:1 ratio to receive oral metformin (2000 mg daily) or observation for 18 months. Fundus autofluorescence imaging was obtained at baseline and every 6 months. Main Outcome Measures: The primary efficacy endpoint was the annualized enlargement rate of the square root-transformed GA area. Secondary endpoints included best-corrected visual acuity (BCVA) and low luminance visual acuity (LLVA) at each visit. Results: Of 66 enrolled participants, 34 (57 eyes) were randomized to the observation group and 32 (53 eyes) were randomized to the treatment group. The median follow-up duration was 13.9 and 12.6 months in the observation and metformin groups, respectively. The mean ± standard error annualized enlargement rate of square root transformed GA area was 0.35 ± 0.04 mm/year in the observation group and 0.42 ± 0.04 mm/year in the treatment group (risk difference = 0.07 mm/year, 95% confidence interval = -0.05 to 0.18 mm/year; P = 0.26). The mean ± standard error decline in BCVA was 4.8 ± 1.7 letters/year in the observation group and 3.4 ± 1.1 letters/year in the treatment group (P = 0.56). The mean ± standard error decline in LLVA was 7.3 ± 2.5 letters/year in the observation group and 0.8 ± 2.2 letters/year in the treatment group (P = 0.06). Fourteen participants in the metformin group experienced nonserious adverse events related to metformin, with gastrointestinal side effects as the most common. No serious adverse events were attributed to metformin. Conclusions: The results of this trial as conducted do not support oral metformin having effects on reducing the progression of GA. Additional placebo-controlled trials are needed to explore the role of metformin for AMD, especially for earlier stages of the disease. Financial Disclosures: Proprietary or commercial disclosure may be found in the Footnotes and Disclosures at the end of this article.

Improving Performance of Al2O3/AlN/GaN MIS HEMTs via In Situ N2 Plasma Annealing.

A novel monocrystalline AlN interfacial layer formation method is proposed to improve the device performance of the fully recessed-gate Al2O3/AlN/GaN Metal-Insulator-Semiconductor High Electron Mobility Transistors (MIS-HEMTs), which is achieved by plasma-enhanced atomic layer deposition (PEALD) and in situ N2 plasma annealing (NPA). Compared with the traditional RTA method, the NPA process not only avoids the device damage caused by high temperatures but also obtains a high-quality AlN monocrystalline film that avoids natural oxidation by in situ growth. As a contrast with the conventional PELAD amorphous AlN, C-V results indicated a significantly lower interface density of states (Dit) in a MIS C-V characterization, which could be attributed to the polarization effect induced by the AlN crystal from the X-ray Diffraction (XRD) and Transmission Electron Microscope (TEM) characterizations. The proposed method could reduce the subthreshold swing, and the Al2O3/AlN/GaN MIS-HEMTs were significantly enhanced with ~38% lower on-resistance at Vg = 10 V. What is more, in situ NPA provides a more stable threshold voltage (Vth) after a long gate stress time, and ΔVth is inhibited by about 40 mV under Vg,stress = 10 V for 1000 s, showing great potential for improving Al2O3/AlN/GaN MIS-HEMT gate reliability.

Blending controlled-release urea and urea under ridge-furrow with plastic film mulching improves yield while mitigating carbon footprint in rainfed potato.

Ridge-furrow with plastic film mulching and various urea types have been applied in rainfed agriculture, but their interactive effects on potato (Solanum tuberosum L.) yield and especially environments remain poorly understood. A three-year experiment was conducted to explore the responses of tuber yield, methane (CH4) and nitrous oxide (N2O) emissions, net global warming potential (NGWP), carbon footprint (CF), and net ecosystem economic budget (NEEB) of rainfed potato to two mulching practices [plastic film mulching (RM) and no plastic film mulching (NM)] and three urea types [conventional urea (U), controlled-release urea (C), and a mixture of equal amounts of conventional urea and controlled-release urea at a ratio of 1:1 (CU)] and their interactions. The results showed that RM significantly decreased cumulative N2O emissions and CH4 uptake by 4.9% and 28.4%, but significantly increased NGWP by 8.9% relative to NM. Compared with U, the C and CU produced much lower cumulative N2O emissions and NGWP and higher CH4 uptake. The interaction of mulching methods and urea type had significant influence on tuber yield and NEEB. Considering both environment and production, RMCU could not only achieve a high tuber yield and NEEB (by up to 26.5% and 42.9%, respectively), but also reduce the CF (by up to 13.7%), and therefore should be considered an effective strategy for dryland potato.

A Fast Transient Adaptive On-Time Controlled BUCK Converter with Dual Modulation.

This paper proposed a fully integrated adaptive on-time (AOT) controlled buck converter with fast load transient. An adaptive on-time generator is presented to stabilize the output frequency. To enhance the light load efficiency, the converter could transfer from the pulse width modulation (PWM) to pulse skip modulation (PSM) as the load current decreases. The buck converter can switch between these two modulation modes adaptively with the assistance of a zero current detection circuit. Implemented in the TSMC 0.18 µm BCD (BiCMOS/DMOS) process, the proposed buck converter works with an input voltage ranging from 5.5 to 15 V, an output voltage ranging from 0.5 to 5 V, and an output load ranging up to 5 A. The experimental results show that based on the dual modulation adaptive on-time controlled mode, the transient recovery time from light to heavy load and from heavy load to light load is 13 µs and 15 µs, respectively. An overshot voltage of 57 mV and an undershot voltage of 53 mV are also achieved.

Associations of systemic health and medication use with the enlargement rate of geographic atrophy in age-related macular degeneration.

BACKGROUND: The associations of geographic atrophy (GA) progression with systemic health status and medication use are unclear. METHODS: We manually delineated GA in 318 eyes in the Age-Related Eye Disease Study. We calculated GA perimeter-adjusted growth rate as the ratio between GA area growth rate and mean GA perimeter between the first and last visit for each eye (mean follow-up=5.3 years). Patients' history of systemic health and medications was collected through questionnaires administered at study enrolment. We evaluated the associations between GA perimeter-adjusted growth rate and 27 systemic health factors using univariable and multivariable linear mixed-effects regression models. RESULTS: In the univariable model, GA perimeter-adjusted growth rate was associated with GA in the fellow eye at any visit (p=0.002), hypertension history (p=0.03), cholesterol-lowering medication use (p<0.001), beta-blocker use (p=0.02), diuretic use (p<0.001) and thyroid hormone use (p=0.03). Among the six factors, GA in the fellow eye at any visit (p=0.008), cholesterol-lowering medication use (p=0.002), and diuretic use (p<0.001) were independently associated with higher GA perimeter-adjusted growth rate in the multivariable model. GA perimeter-adjusted growth rate was 51.1% higher in patients with versus without cholesterol-lowering medication use history and was 37.8% higher in patients with versus without diuretic use history. CONCLUSIONS: GA growth rate may be associated with the fellow eye status, cholesterol-lowering medication use, and diuretic use. These possible associations do not infer causal relationships, and future prospective studies are required to investigate the relationships further.

ABC Transporters and CYP3A4 Mediate Drug Interactions between Enrofloxacin and Salinomycin Leading to Increased Risk of Drug Residues and Resistance.

Enrofloxacin (ENR) is one of the most common drugs used in poultry production to treat bacterial diseases, and there is a high risk of drug interactions (DDIs) between polyether anticoccidial drugs added to poultry feed over time. This may affect the efficacy of antibiotics or lead to toxicity, posing a potential risk to the environment and food safety. This study aimed to investigate the DDI of ENR and salinomycin (SAL) in broilers and the mechanism of their DDI. We found that SAL increased the area under the curve and elimination half-life of ENR and ciprofloxacin (CIP) by 1.3 and 2.4 times, 1.2 and 2.5 times, respectively. Cytochrome 3A4 (CYP3A4), p-glycoprotein (P-gp) and breast cancer resistance protein (BCRP) were important factors for the DDI between ENR and SAL in broilers. ENR and SAL are substrates of CYP3A4, P-gp and BCRP in broilers; ENR and SAL inhibited the expression of CYP3A4 activity in a time- and concentration-dependent. Meanwhile, ENR downregulated the expression of P-gp and BCRP in a time- and concentration-dependent manner. A single oral administration of SAL inhibited CYP3A4, P-gp, and BCRP, but long-term mixed feeding upregulated the expression of CYP3A4, P-gp, and BCRP. Molecular docking revealed that ENR and SAL compete with each other for CYP3A4 to affect hepatic metabolism, and compete with ATP for P-gp and BCRP binding sites to inhibit efflux. ENR and SAL in broilers can lead to severe DDI. Drug residues and resistance following co-administration of ENR and SAL and other SAL-based drug-feed interactions warrant further study.

Identification and Expression Analysis of DFR Gene Family in Brassica napus L.

Dihydroflavonol 4-reductase (DFR) is a key enzyme in the flavonoid biosynthetic pathway and is essential for the formation of plants' color. In this study, 26 BnDFR genes were identified using 6 Arabidopsis DFR genes as reference. The physicochemical properties, subcellular localization, and conserved structure of BnDFR proteins were analyzed; the evolutionary relationship, collinearity analysis, and expression characteristics of BnDFR genes were studied; and the correlation between the expression level of BnDFR genes and anthocyanin content in rape petals were analyzed. The results showed that the 26 BnDFRs were located in chloroplasts, cytoplasm, nuclei, and mitochondria, distributed on 17 chromosomes, and divided into 4 groups; members of the same group have a similar function, which may be related to the environmental response elements and plant hormone response elements. Intraspecific collinearity analysis showed 51 pairs of collinear genes, and interspecific collinearity analysis showed 30 pairs of collinear genes. Analysis of the expression levels of BnDFRs and anthocyanin content in different color rape petals showed that BnDFR6 and BnDFR26 might play an important role in the synthesis of anthocyanins in rape petals. This provides theoretical guidance for further analysis of the anthocyanin anabolism mechanism involved in the DFR gene in Brassica napus.

Integration of mRNA and miRNA analysis reveals the molecular mechanisms of sugar beet (Beta vulgaris L.) response to salt stress.

The continuous increase of saline-alkali areas worldwide has led to the emergence of saline-alkali conditions, which are the primary abiotic stress or hindering the growth of plants. Beet is among the main sources of sugar, and its yield and sugar content are notably affected by saline-alkali stress. Despite sugar beet being known as a salt-tolerant crop, there are few studies on the mechanisms underlying its salt tolerance, and previous studies have mainly delineated the crop's response to stress induced by NaCl. Recently, advancements in miRNA-mRNA network analysis have led to an increased understanding of how plants, including sugar beet, respond to stress. In this study, seedlings of beet variety "N98122" were grown in the laboratory using hydroponics culture and were exposed to salt stress at 40 days of growth. According to the phenotypic adaptation of the seedlings' leaves from a state of turgidity to wilting and then back to turgidity before and after exposure, 18 different time points were selected to collect samples for analysis. Subsequently, based on the data of real-time quantitative PCR (qRT-PCR) of salt-responsive genes, the samples collected at the 0, 2.5, 7.5, and 16 h time points were subjected to further analysis with experimental materials. Next, mRNA-seq data led to the identification of 8455 differentially expressed mRNAs (DEMs) under exposure to salt stress. In addition, miRNA-seq based investigation retrieved 3558 miRNAs under exposure to salt stress, encompassing 887 known miRNAs belonging to 783 families and 2,671 novel miRNAs. With the integrated analysis of miRNA-mRNA network, 57 miRNA-target gene pairs were obtained, consisting of 55 DEMIs and 57 DEMs. Afterwards, we determined the pivotal involvement of aldh2b7, thic, and δ-oat genes in the response of sugar beet to the effect of salt stress. Subsequently, we identified the miRNAs novel-m035-5p and novel-m0365-5p regulating the aldh gene and miRNA novel-m0979-3p regulating the thic gene. The findings of miRNA and mRNA expression were validated by qRT-PCR.

Natural history of central sparing in geographic atrophy secondary to non-exudative age-related macular degeneration.

BACKGROUND: The macular central 1 mm diameter zone is crucial to patients' visual acuity, but the long-term natural history of central sparing in eyes with geographic atrophy (GA) is unknown. METHODS: We manually segmented GA in 210 eyes with GA involving central 1 mm diameter zone (mean follow-up=3.8 years) in the Age-Related Eye Disease Study. We measured the residual area in central 1 mm diameter zone and calculated central residual effective radius (CRER) as square root of (residual area/π). A linear mixed-effects model was used to model residual size over time. We added a horizontal translation factor to each data set to account for different durations of GA involving the central zone. RESULTS: The decline rate of central residual area was associated with baseline residual area (p=0.008), but a transformation from central residual area to CRER eliminated this relationship (p=0.51). After the introduction of horizontal translation factors to each data set, CRER declined linearly over approximately 13 years (r2=0.80). The growth rate of total GA effective radius was 0.14 mm/year (95% CI 0.12 to 0.15), 3.7-fold higher than the decline rate of CRER (0.038 mm/year, 95% CI 0.034 to 0.042). The decline rate of CRER was 53.3% higher in eyes with than without advanced age-related macular degeneration in the fellow eyes at any visit (p=0.007). CONCLUSIONS: CRER in eyes with GA declined linearly over approximately 13 years and may serve as an anatomic endpoint in future clinical trials aiming to preserve the central zone.

Relationship between gut microbiota and lymphocyte subsets in Chinese Han patients with spinal cord injury.

This study is to investigate the changes of lymphocyte subsets and the gut microbiota in Chinese Han patients with spinal cord injury (SCI). We enrolled 23 patients with SCI and 21 healthy controls. Blood and fecal samples were collected. The proportion of lymphocyte subsets was detected by flow cytometry. 16S rDNA sequencing of the V4 region was used to analyze the gut microbiota. The changes of the gut microbiota were analyzed by bioinformatics. Correlation analysis between gut microbiota and lymphocyte subsets was performed. CD4 + cells, CD4 + /CD8 + ratio and CD4 + CD8 + cells in peripheral blood of SCI patients were significantly lower than those of the control group (P < 0.05). There was no significant difference in B cells and CIK cells between the SCI group and the control group. The gut microbiota community diversity index of SCI patients was significantly higher than that of healthy controls. In SCI patients, the relative abundance of Lachnospiraceae (related to lymphocyte subset regulation), Ruminococcaceae (closely related to central nervous system diseases), and Escherichia-Shigella (closely related to intestinal infections) increased significantly, while the butyrate producing bacteria (Fusobacterium) that were beneficial to the gut were dramatically decreased. Correlation analysis showed that the five bacterial genera of SCI patients, including Lachnospiraceae UCG-008, Lachnoclostridium 12, Tyzzerella 3, Eubacterium eligens group, and Rumencocciucg-002, were correlated with T lymphocyte subsets and NK cells. In the SCI group, the flora Prevotella 9, Lachnospiraceae NC2004 group, Veillonella, and Sutterella were positively correlated with B cells. However, Fusobacterium and Akkermansia were negatively correlated with B cells. Moreover, Roseburia and Ruminococcaceae UCG-003 were positively correlated with CIK cells. Our results suggest that the gut microbiota of patients with SCI is associated with lymphocyte subsets. Therefore, it is possible to improve immune dysregulation in SCI patients by modulating gut microbiota, which may serve as a new therapeutic method for SCI.