The Impact of Cataracts on the Measurement of Macular Choriocapillaris Flow Deficits Using Swept-Source OCT Angiography.

Purpose: The impact of cataracts on the measurement of macular choriocapillaris flow deficits (CC FDs) was assessed by comparing the quantitative results before and after cataract surgery using an image quality algorithm developed for swept-source optical coherence tomography angiography (SS-OCTA) scans and a validated strategy for quantifying the CC FDs. Methods: SS-OCTA image quality scores and CC FDs measurements within the fovea-centered 1-mm, 3-mm, and 5-mm diameter circles were compared before and after cataract surgery. CC FDs changes in a modified Early Treatment Diabetic Retinopathy Study (ETDRS) grid were further investigated. Results: Twenty-four eyes were studied. Overall image quality in all three circles was observed to improve significantly following the removal of cataracts (all P < 0.05). Although there was good repeatability in the measurements of CC FDs at both visits (intraclass correlation coefficients were over 0.95), significant decreases in CC FD measurements were observed after surgery within the 1-mm circle (P < 0.001) and the 3-mm circle (P = 0.011), but no changes were observed within the 5-mm circle (P = 0.509) or any of the quadrant sectors of the modified ETDRS grid (all P > 0.05). Conclusions: The presence of cataracts resulted in worse image quality and increased CC FD measurements within the fovea-centered 1-mm and 3-mm circles, with the 1-mm circle being impacted the most. Translational Relevance: The impaired detection of CC perfusion deficits within the central macula of cataract eyes needs to be appreciated when imaging the CC in phakic eyes, especially in clinical trials.

Biochar and nano-ferric oxide synergistically alleviate cadmium toxicity of muskmelon.

Cadmium is toxic to plants. The accumulation of cadmium in edible plants such as muskmelon may affect the safe production of crops and result in human health problem. Thus effective measures are urgently needed for soil remediation. This work aims to investigate the effects of nano-ferric oxide and biochar alone or mixture on muskmelon under cadmium stress. The results of growth and physiological indexes showed that compared with the application of cadmium alone, the composite treatment (biochar and nano-ferric oxide) decreased malondialdehyde content by 59.12% and ascorbate peroxidase activity increased by 276.6%. Their addition can increase the stress resistance of plants. The results of soil analysis and cadmium content determination in plants showed that the composite treatment was beneficial to reduce the cadmium content in various parts of muskmelon. In the presence of high concentration of cadmium, the Target Hazard Quotient value of peel and flesh of muskmelon in the composite treatment was less than 1, which means the edible risk was greatly reduced. Furthermore, the addition of composite treatment increased the content of effective components; the contents of polyphenols, flavonoids, and saponins in the flesh of the compound treatment were increased by 99.73%, 143.07%, and 18.78% compared with the cadmium treatment. The results provide a technical reference for the further application of biochar combined with nano-ferric oxide in the field of soil heavy metal remediation, and provide a theoretical basis for further research on reducing the toxicity of cadmium to plants and improving the edible quality of crops.

Transcriptome studies on cadmium tolerance and biochar mitigating cadmium stress in muskmelon.

Cadmium pollution in agricultural soil is a great threat to crop growth and human health. In this research, with 1%, 3% and 5% biochar applied to control soil cadmium pollution, melon was selected to be the experimental object for physiological detection and transcriptome analysis, through which we explored the mechanism of cadmium tolerance and biochar mitigating cadmium stress in muskmelon. Three set concentrations of biochar have a mitigative effect on muskmelon cadmium stress, and 5% biochar and 3% biochar respectively have the best and the worst alleviative effect. The alleviation of biochar to cadmium stress on muskmelon is primarily in the manner of inhibiting cadmium transfer, while the resistance of muskmelon to cadmium stress is through activating phenylpropanoid pathway and overexpressing stress related genes. Under cadmium treatment, 11 genes of the phenylpropane pathway and 19 stress-related genes including cytochrome P450 family protein genes and WRKY transcription factor genes were up-regulated, while 1%, 3%, 5% biochar addition significantly downregulated 3, 0, 7 phenylpropane pathway genes and 17, 5, 16 stress-related genes, respectively. Genes such as cytochrome P450 protein family genes, WRKY transcription factor genes, and annexin genes may play a key role in muskmelon's resistance to cadmium stress. The results show the key pathways and genes of cadmium stress resistance and the effect of different concentrations of biochar in alleviating cadmium stress, which provide a reference for the research of cadmium stress resistance in crops and the application of biochar in cadmium pollution in agricultural soil.

Investigating the AC079305/DUSP1 Axis as Oxidative Stress-Related Signatures and Immune Infiltration Characteristics in Ischemic Stroke.

Background: Oxidative stress (OS) and immune inflammation play complex intersections in the pathophysiology of ischemic stroke (IS). However, a competing endogenous RNA- (ceRNA-) based mechanism linked to the intersections in IS has not been explored. We aimed to identify potential OS-related signatures and analyze immune infiltration characteristics in IS. Methods: Three datasets (GSE22255, GSE110993, and GSE140275) from the GEO database were extracted. Differentially expressed long noncoding RNAs, microRNAs, and messenger RNAs (DElncRNAs, DEmiRNAs, and DEmRNAs) between IS patients and controls were identified. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis were explored. Moreover, a triple ceRNA network was constructed to reveal transcriptional regulation mechanisms. A comprehensive strategy among least absolute shrinkage and selection operator (LASSO) regression, DEmRNAs, uprelated DEmRNAs, and OS-related genes was adopted to select the best signature. Then, we evaluated and verified the discriminant ability of the signature via receiver operating characteristic (ROC) analysis. Immune infiltration characteristics were explored via the CIBERSORT algorithm. Moreover, the best signature was verified via qPCR and western blot methods in rat brain tissues and PC12 cells. Results: 11 DEmRNAs were identified totally. Enrichment analysis showed that the DEmRNAs were primarily concentrated in MAPK-associated biological processes and immune or inflammation-involved pathways. DUSP1 was identified as the best signature with an area under the ROC curve of 73.5% (95%CI = 57.02-89.98, sensitivity = 95%, and specificity = 60%) in GSE22255 and 100.0% (95%CI = 100.00-100.00, sensitivity = 100%, and specificity = 100%) in GSE140275. Importantly, we also identified the AC079305/DUSP1 axis in the ceRNA network. Immune infiltration showed that resting mast cells infiltrate less in IS patients compared with controls. And DUSP1 was negatively correlated with resting mast cells (r = -0.703, P < 0.01), whereas it was positively correlated with neutrophils (r = 0.339, P < 0.05). Both in vivo and in vitro models confirmed the upregulated expression of DUSP1 and the downregulated expression of miR-429. Conclusion: This study identified the ceRNA-based AC079305/DUSP1 axis as a promising OS-related signature for IS. Immune infiltrating cells, especially mast cells, may exert a pivotal role in IS progression. Pharmacological agents targeting signatures, their receptors, or mast cells may shed a novel light on therapeutic targets for IS.

Application of OCT-Derived Attenuation Coefficient in Acute Burn-Damaged Skin.

BACKGROUND AND OBJECTIVES: There remains a need to objectively monitor burn wound healing within a clinical setting, and optical coherence tomography (OCT) is proving itself one of the ideal modalities for just such a use. The aim of this study is to utilize the noninvasive and multipurpose capabilities of OCT, along with its cellular-level resolution, to demonstrate the application of optical attenuation coefficient (OAC), as derived from OCT data, to facilitate the automatic digital segmentation of the epidermis from scan images and to work as an objective indicator for burn wound healing assessment. STUDY DESIGN/MATERIALS AND METHODS: A simple, yet efficient, method was used to estimate OAC from OCT images taken over multiple time points following acute burn injury. This method enhanced dermal-epidermal junction (DEJ) contrast, which facilitated the automatic segmentation of the epidermis for subsequent thickness measurements. In addition, we also measured and compared the average OAC of the dermis within said burns for correlative purposes. RESULTS: Compared with unaltered OCT maps, enhanced DEJ contrast was shown in OAC maps, both from single A-lines and completed B-frames. En face epidermal thickness and dermal OAC maps both demonstrated significant changes between imaging sessions following burn injury, such as a loss of epidermal texture and decreased OAC. Quantitative analysis also showed that OAC of acute burned skin decreased below that of healthy skin following injury. CONCLUSIONS: Our study has demonstrated that the OAC estimated from OCT data can be used to enhance imaging contrast to facilitate the automatic segmentation of the epidermal layer, as well as help elucidate our understanding of the pathological changes that occur in human skin when exposed to acute burn injury, which could serve as an objective indicator of skin injury and healing.

Salt-Dependent RNA Pseudoknot Stability: Effect of Spatial Confinement.

Macromolecules, such as RNAs, reside in crowded cell environments, which could strongly affect the folded structures and stability of RNAs. The emergence of RNA-driven phase separation in biology further stresses the potential functional roles of molecular crowding. In this work, we employed the coarse-grained model that was previously developed by us to predict 3D structures and stability of the mouse mammary tumor virus (MMTV) pseudoknot under different spatial confinements over a wide range of salt concentrations. The results show that spatial confinements can not only enhance the compactness and stability of MMTV pseudoknot structures but also weaken the dependence of the RNA structure compactness and stability on salt concentration. Based on our microscopic analyses, we found that the effect of spatial confinement on the salt-dependent RNA pseudoknot stability mainly comes through the spatial suppression of extended conformations, which are prevalent in the partially/fully unfolded states, especially at low ion concentrations. Furthermore, our comprehensive analyses revealed that the thermally unfolding pathway of the pseudoknot can be significantly modulated by spatial confinements, since the intermediate states with more extended conformations would loss favor when spatial confinements are introduced.

Robust three-dimensional registration on optical coherence tomography angiography for speckle reduction and visualization.

BACKGROUND: In the clinical applications of optical coherence tomography angiography (OCTA), the repeated scanning and averaging method can provide better contrast with reduced speckle noises in the final results, which are useful for visualizing and quantifying vascular components with high accuracy, reproducibility, and reliability. However, the inevitable patient motion presents a challenge to this method. The objective of this study is to meet this challenge by introducing a 3D registration method to register optical coherence tomography (OCT)/OCTA scans for precise volume averaging of multiple scans to improve the signal-to-noise ratio (SNR) and increase quantification accuracy. METHODS: The proposed method utilized both rigid affine transformation and non-rigid B-spline transformation in which their parameters were optimized and calculated by the average stochastic gradient descent on OCT structural images. In addition, we also introduced a multi-level resolution approach to further improve the robustness and computational speed of our proposed method. The imaging performance was tested on in vivo imaging of human skin and eye and assessed by SNR, peak signal-to-noise ratio (PSNR) and normalized correlation coefficient (NCC). RESULTS: Five subjects were enrolled in this study for obtaining in vivo images of skin and retina. The proposed registration and averaging method provided substantial improvements of the imaging performance in terms of vessel connectivity and signal to noise ratio. The increase of repeated volume numbers in the averaging improves all the metrics assessed, i.e., SNR, PSNR and NCC. An improvement of the SNR from 10 to 40 dB after 10 repeated volumetric averaging was achieved. CONCLUSIONS: The proposed 3D registration and averaging method is effective in reducing speckle noises and suppressing motion artifacts, thereby improving SNR, PSNR and NCC metrics for final averaged images. It is expected that the proposed algorithm would be practically useful in better visualization and more reliable quantification of in vivo OCT and OCTA data, which would be beneficial to OCT clinical applications.

Optical coherence tomography angiography measures blood pulsatile waveforms at variable tissue depths.

BACKGROUND: Photoplethysmography (PPG) is routinely used to detect the blood pulse signal from skin tissue beds in clinics. However, the origin of the PPG signal remains controversial. The purpose of this study is to explore optical coherence tomography angiography (OCTA) to indicate pulsatile waveforms in the papillary plexus and dermal plexus separately under different hand elevations. METHOD: Optical microangiography (OMAG) algorithm was used to obtain a 3D OCTA signals, from which the depth-resolved pulsatile blood flow signals were extracted from different skin vascular plexus. The systolic amplitude, crest time, and delta T were measured from the OCTA pulsatile signals when the hand was placed at the positions of 50 cm below, 0 cm, and 50 cm above the heart level. RESULTS: The pulse signal integrated from all the depths has a similar waveform to that of the PPG and showed the same morphological change at different hand elevations. The pulsatile patterns from the papillary plexus and dermal plexus showed distinct morphological changes at different local blood pressure. Less amplitude difference was found from papillary plexus comparing to that of the dermal plexus. Crest time was found in an increasing trend in the OCTA pulsatile waveform from both plexuses when the arm was raised from the position below to above the heart level. In contrast, a decreasing trend of Delta T was detected in the dermal pulsatile but was not observed from that of the papillary plexus, indicating that vascular resistance associated with the arm elevations does not necessarily have the same effect on the two plexuses. CONCLUSIONS: OCTA can provide depth-resolved pulsatile waveforms within different microvascular plexus within tissue skin beds. This technique could open doors to understanding the mechanisms of how blood flow changes at different skin circulatory plexus.

Optical coherence tomography angiography distortion correction in widefield montage images.

BACKGROUND: Optical coherence tomography (OCT) imaging is inherently susceptible to distortion artifacts due to the natural curvature of the eye. This study proposes a novel model for widefield OCT angiography (OCTA) distortion correction and analyzes the effects of this correction on quantification metrics. METHODS: Widefield OCTA images were obtained on normal subjects at five fixation spatial positions. Radial and field distortion correction were applied and images stitched together to form a corrected widefield montage image. Vessel area density (VAD), vessel complexity index (VCI), and flow impairment area were quantified on the original and corrected montage images. RESULTS: This model allows for distortion correction and montaging of widefield images. There were either statistically insignificant or small magnitude changes in vessel density and vessel complexity between uncorrected and corrected widefield images. There was a significant and large difference in flow impairment area, both in the macular (+8.2%, P=0.049) and peripheral areas (+17.2%, P=0.011), following correction. The relationship between pre- and post-correction flow impairment area was non-linear. CONCLUSIONS: Distortion correction of widefield OCTA images can result in clinically and statistically significant differences in important quantification metrics. This effect appears to be most pronounced in the periphery.

Scaffold-free human mesenchymal stem cell construct geometry regulates long bone regeneration.

Biomimetic bone tissue engineering strategies partially recapitulate development. We recently showed functional restoration of femoral defects using scaffold-free human mesenchymal stem cell (hMSC) condensates featuring localized morphogen presentation with delayed in vivo mechanical loading. Possible effects of construct geometry on healing outcome remain unclear. Here, we hypothesized that localized presentation of transforming growth factor (TGF)-ß1 and bone morphogenetic protein (BMP)-2 to engineered hMSC tubes mimicking femoral diaphyses induces endochondral ossification, and that TGF-ß1 + BMP-2-presenting hMSC tubes enhance defect healing with delayed in vivo loading vs. loosely packed hMSC sheets. Localized morphogen presentation stimulated chondrogenic priming/endochondral differentiation in vitro. Subcutaneously, hMSC tubes formed cartilage templates that underwent bony remodeling. Orthotopically, hMSC tubes stimulated more robust endochondral defect healing vs. hMSC sheets. Tissue resembling normal growth plate was observed with negligible ectopic bone. This study demonstrates interactions between hMSC condensation geometry, morphogen bioavailability, and mechanical cues to recapitulate development for biomimetic bone tissue engineering.

OCT-Based Angiography and Surface Topography in Burn-Damaged Skin.

BACKGROUND AND OBJECTIVES: There is a clinical need for an accurate, non-invasive imaging tool that can provide the objective assessment of burn wounds. The aims of this study are to demonstrate the potential of optical coherence tomography (OCT) in evaluating burn wound healing, as well as exploring the physiological basis of human wound healing. STUDY DESIGN/MATERIALS AND METHODS: This was a retrospective study. Seven patients with severe burn wounds who were admitted to Harborview Medical Center were imaged using an in-house-built, clinical-prototype OCT system. OCT imaging was carried out at multiple scan sites on the burned skin across two time points (imaging session #1 and #2) with a field of view of ~9 × 9 mm. Due to pathological differences among burn zones, scan sites were classified into red sites (zone of hyperemia), white sites (zone of coagulation), and mixed sites. In addition to obtaining qualitative en face vascular and surface topography maps, we quantified vascular area density and surface roughness for comparative purposes. RESULTS: En face vascular and surface topography maps demonstrated numerous morphological changes over both imaging sessions associated with burn injury, such as altered blood flow and loss of regular texture. Quantitative analyses revealed that during imaging session #1, vascular area density was significantly increased within the red sites compared with that of a healthy control (P = 0.0130), while vascular area density was significantly decreased within the white sites compared with that of a healthy control (P < 0.0001). During imaging session #2, vascular area density was significantly reduced to a more normal range within the red sites compared with imaging session #1 (P = 0.0215); however, vascular area density was still significantly lower within the white sites compared with that of a healthy control (P < 0.0001). Furthermore, vascular area density and surface roughness were significantly increased within the white sites during imaging session #2 compared with imaging session #1 (both P < 0.0001). CONCLUSIONS: OCT is clinically feasible to monitor vascular changes and alterations in skin surface roughness during the process of burn wound healing. Variations in vascular area density and roughness measurements within the burn wounds revealed by OCT offer some key insights into the underlying pathophysiological mechanisms responsible for wound healing, which may become critical biological indicators in future clinical evaluation and monitoring of wound healing. Lasers Surg. Med. © 2020 Wiley Periodicals LLC.

Applications of carbon quantum dots to alleviate Cd2+ phytotoxicity in Citrus maxima seedlings.

Heavy metal pollution may affect plant growth. The focus of this study was to explore remediation agents that alleviate cadmium toxicity in plants. The Citrus maxima (grapefruit) seedlings were cultivated for 10 days under hydroponic conditions amended with different concentrations of Cd2+ (50 and 200 mg/L) and CDs (600 and 900 mg/L). Our observations on roots and leaves showed that, the plant exposed to 200 mg/L Cd2+ alone was damaged, supported by the changes in anthocyanin contents, activity of antioxidant enzymes and cell membrane peroxidation damage (up to 35.8-45%). However, the physiological properties of the plant were improved upon exposed to 200 mg/L Cd2+ plus 900 mg/L CDs; it can be ascribed to Cd2+ sorption to the co-exposed CDs which reduced its freely dissolved concentration by more than 22.5%, thus significantly reducing the amount of Cd2+ entered the plant roots by 50.7-89.4%. Due to the oxidative stress induced by Cd2+ exposure at 200 mg/L, expression of glutathione-producing genes was up-regulated by 30-360% relative to the control, while the genes expression upon exposure to 200 mg/L Cd2+ and 900 mg/L CDs was reduced by 48.4-91.5% relative to that exposed to 200 mg/L Cd2+ alone. However, detoxification of CDs on plant leaves at 600 mg/L was insignificant, because a portion of Cd2+ taken up by roots can be transported to leaves associated with the internalized CDs. Therefore, CDs can be utilized as a repair agent to mitigate toxicity of Cd2+ to plant especially at a high amendment level (900 mg/L).

Compound repair effect of carbon dots and Fe2+ on iron deficiency in Cucumis melon L.

Iron-deficiency is one of the most widespread micronutrient deficiency faced by plants, and proper iron supplementation is essential for the growth of crops and for people to obtain iron from food. In order to explore new methods of iron supplementation, we studied the repair effect of CDs on iron-deficient (Cucumis melo L.) muskmelon. Iron-deficient muskmelons were treated with different concentrations of Fe2+, CDs and their complexes. The results showed that CDs significantly increased the iron transport rate and it is noteworthy that 75 mg/L CDs increased the iron transport rate of 0.7 mg/L Fe2+ by 134%. The compound treatment reduced the oxidative stress caused by iron deficiency, such as the CAT activity in the leaves of the compound treatment group was 10%-50% lower than that of the iron supplementation alone. Fluorescent imaging results of melon proved that CDs entered into the muskmelon seedlings. In combination with the above results and the adsorption of CDs, we speculated that the way CDs promoted iron absorption and transport was most likely to combine with Fe2+ and co-transport in melon, which changed the content of reactive oxygen species and other free radicals, thus causing changes of physiological state of melon. This study confirmed that CDs had a positive effect on the iron deficiency of muskmelon, and improved the growth of muskmelon under the condition of iron deficiency, which has a certain reference value for further optimization of iron supplementation solution.

Thiol-Epoxy "Click" Chemistry to Engineer Cytocompatible PEG-Based Hydrogel for siRNA-Mediated Osteogenesis of hMSCs.

Thiol-epoxy "click" chemistry is employed for the first time to engineer a new cytocompatible PEG-based hydrogel system in aqueous media with the ability to encapsulate human mesenchymal stem cells (hMSCs) and control their fate for tissue regeneration. Cells were easily encapsulated into the hydrogels and exhibited high cell viability over 4 weeks of culture regardless of the presence of siRNA, complexed with polyethylenimine (PEI) in the form of siRNA/PEI nanocomplexes, indicating the biocompatibility of the developed hydrogel. Loading pro-osteogenic siNoggin in the hydrogel significantly enhanced the osteogenesis of encapsulated hMSCs, demonstrating the potential application of this system in tissue engineering.