Functional OCT reveals anisotropic changes of retinal flicker-evoked vasodilation.

The purpose of this study is to verify the effect of anisotropic property of retinal biomechanics on vasodilation measurement. A custom-built optical coherence tomography (OCT) was used for time-lapse imaging of flicker stimulation-evoked vessel lumen changes in mouse retinas. A comparative analysis revealed significantly larger (18.21%) lumen dilation in the axial direction compared to the lateral (10.77%) direction. The axial lumen dilation predominantly resulted from the top vessel wall movement toward the vitreous direction, whereas the bottom vessel wall remained stable. This observation indicates that the traditional vasodilation measurement in the lateral direction may result in an underestimated value.

Intrinsic signal optoretinography of dark adaptation abnormality due to rod photoreceptor degeneration.

This research aims to investigate the potential of using intrinsic optical signal (IOS) optoretinography (ORG) to objectively detect dark adaptation (DA) abnormalities related to rod photoreceptor degeneration. Functional optical coherence tomography (OCT) was employed in both wild-type (WT) and retinal degeneration 10 (rd10) mice to conduct this assessment. Dynamic OCT measurements captured the changes in retinal thickness and reflectance from light-to-dark transition. Comparative analysis revealed significant IOS alterations within the outer retina. Specifically, a reduction in thickness from external limiting membrane (ELM) peak to retinal pigment epithelium (RPE) peak was observed (WT: 1.13 ± 0.69 µm, 30 min DA; rd10: 2.64 ± 0.86 µm, 30 min DA), as well as a decrease in the intensity of the inner segment ellipsoid zone (EZ) in 30 min DA compared to light adaptation (LA). The reduction of relative EZ intensity was notable in rd10 after 5 min DA and in WT after 15 min DA, with a distinguishable difference between rd10 and WT after 10 min DA. Furthermore, our findings indicated a significant decrease in the relative intensity of the hypo-reflective band between EZ and RPE in rd10 retinas during DA, which primarily corresponds to the outer segment (OS) region. In conclusion, the observed DA-IOS abnormalities, including changes in ELM-RPE thickness, EZ, and OS intensity, hold promise as differentiators between WT and rd10 mice before noticeable morphological abnormalities occur. These findings suggest the potential of this non-invasive imaging technique for the early detection of dysfunction in retinal photoreceptors.

Intrinsic signal optoretinography revealing AD-induced retinal photoreceptor hyperexcitability before a detectable morphological abnormality.

Neuronal hyperexcitability promises an early biomarker of Alzheimer's disease (AD). However, in vivo detection of neuronal hyperexcitability in the brain is technically challenging. The retina, one part of the central nervous system, presents a unique window for noninvasive monitoring of the brain function. This study aims to test the feasibility of using intrinsic signal optoretinography (ORG) for mapping retinal hyperexcitability associated with early-stage AD. Custom-designed optical coherence tomography (OCT) was employed for both morphological measurement and functional ORG of wild-type mice and 3xTg-AD mice. Comparative analysis revealed AD-induced retinal photoreceptor hyperexcitability prior to detectable structural degeneration.

Improvement of retinal function in Alzheimer disease-associated retinopathy by dietary lysophosphatidylcholine-EPA/DHA.

Alzheimer disease (AD) is the most prevalent cause of dementia in the elderly. Although impaired cognition and memory are the most prominent features of AD, abnormalities in visual functions often precede them, and are increasingly being used as diagnostic and prognostic markers for the disease. Retina contains the highest concentration of the essential fatty acid docosahexaenoic acid (DHA) in the body, and its deficiency is associated with several retinal diseases including diabetic retinopathy and age related macular degeneration. In this study, we tested the hypothesis that enriching retinal DHA through a novel dietary approach could ameliorate symptoms of retinopathy in 5XFAD mice, a widely employed model of AD. The results show that 5XFAD mice have significantly lower retinal DHA compared to their wild type littermates, and feeding the lysophosphatidylcholine (LPC) form of DHA and eicosapentaenoic acid (EPA) rapidly normalizes the DHA levels, and increases retinal EPA by several-fold. On the other hand, feeding similar amounts of DHA and EPA in the form of triacylglycerol had only modest effects on retinal DHA and EPA. Electroretinography measurements after 2 months of feeding the experimental diets showed a significant improvement in a-wave and b-wave functions by the LPC-diet, whereas the TAG-diet had only a modest benefit. Retinal amyloid ß levels were decreased by about 50% by the LPC-DHA/EPA diet, and by about 17% with the TAG-DHA/EPA diet. These results show that enriching retinal DHA and EPA through dietary LPC could potentially improve visual abnormalities associated with AD.

Rotational Distortion and Compensation in Optical Coherence Tomography with Anisotropic Pixel Resolution.

Accurate image registration is essential for eye movement compensation in optical coherence tomography (OCT) and OCT angiography (OCTA). The spatial resolution of an OCT instrument is typically anisotropic, i.e., has different resolutions in the lateral and axial dimensions. When OCT images have anisotropic pixel resolution, residual distortion (RD) and false translation (FT) are always observed after image registration for rotational movement. In this study, RD and FT were quantitively analyzed over different degrees of rotational movement and various lateral and axial pixel resolution ratio (RL/RA) values. The RD and FT provide the evaluation criteria for image registration. The theoretical analysis confirmed that the RD and FT increase significantly with the rotation degree and RL/RA. An image resizing assisting registration (RAR) strategy was proposed for accurate image registration. The performance of direct registration (DR) and RAR for retinal OCT and OCTA images were quantitatively compared. Experimental results confirmed that unnormalized RL/RA causes RD and FT; RAR can effectively improve the performance of OCT and OCTA image registration and distortion compensation.

Adaptive vessel tracing and segmentation in OCT enables the robust detection of wall-to-lumen ratio abnormalities in 5xFAD mice.

The wall-to-lumen ratio (WLR) of retinal blood vessels promises a sensitive marker for the physiological assessment of eye conditions. However, in vivo measurement of vessel wall thickness and lumen diameter is still technically challenging, hindering the wide application of WLR in research and clinical settings. In this study, we demonstrate the feasibility of using optical coherence tomography (OCT) as one practical method for in vivo quantification of WLR in the retina. Based on three-dimensional vessel tracing, lateral en face and axial B-scan profiles of individual vessels were constructed. By employing adaptive depth segmentation that adjusts to the individual positions of each blood vessel for en face OCT projection, the vessel wall thickness and lumen diameter could be reliably quantified. A comparative study of control and 5xFAD mice confirmed WLR as a sensitive marker of the eye condition.

The complete plastome sequence of Heuchera micrantha Douglas ex Lindl. (Saxifragaceae), an ornamental plant.

The perennial herb Heuchera micrantha (Saxifragaceae) is a popular ornamental plant. However, the plastome sequence of H. micrantha has not been reported yet. Here, we assembled the complete plastome of H. micrantha using Illumina high-throughput pair-end sequencing. The plastome is a circular DNA molecule of 155,469 bp, comprising a pair of inverted repeat (IR, 25,654 bp) regions, a small single copy (SSC, 18,050 bp) region, and a large single copy (LSC, 86,111 bp) region. It encodes 129 genes, of which 84 are protein-coding genes, 37 are transfer RNAs, and eight are rRNAs. The total GC content is 37.8%. Phylogenetic analysis shows that H. micrantha, together with three other Heuchera species is clustered with Tiarella cordifolia. This complete plastome is beneficial for future genetic research on the Heuchera group.

Characterization of the complete chloroplast genome of the Helleborus atrorubens Waldst. & Kit. (Ranunculaceae).

Helleborus atrorubens is an economically important perennial garden plant with medicinal value. Here, we sequenced the complete chloroplast genome of H. atrorubens. The results revealed the chloroplast genome to be 166,695 bp in length. It possesses a typical quadripartite structure containing one large single copy (LSC) region (84994 bp), one small single copy (SSC) region (17,825 bp), and a pair of inverted repeat (IR) regions (31938 bp). This chloroplast genome encoded 130 genes, out of which 85 code for proteins, 37 for transfer RNAs, and 8 for ribosomal RNAs. Simple sequence repeat (SSR) markers and the top variable coding regions were identified. Our study lays a foundation for further research, such as species differentiation and phylogenetic reconstruction of the Helleborus genus.

ADC-Net: An Open-Source Deep Learning Network for Automated Dispersion Compensation in Optical Coherence Tomography.

Chromatic dispersion is a common problem to degrade the system resolution in optical coherence tomography (OCT). This study is to develop a deep learning network for automated dispersion compensation (ADC-Net) in OCT. The ADC-Net is based on a modified UNet architecture which employs an encoder-decoder pipeline. The input section encompasses partially compensated OCT B-scans with individual retinal layers optimized. Corresponding output is a fully compensated OCT B-scan with all retinal layers optimized. Two numeric parameters, i.e., peak signal to noise ratio (PSNR) and structural similarity index metric computed at multiple scales (MS-SSIM), were used for objective assessment of the ADC-Net performance and optimal values of 29.95 ± 2.52 dB and 0.97 ± 0.014 were obtained respectively. Comparative analysis of training models, including single, three, five, seven and nine input channels were implemented. The mode with five-input channels was observed to be optimal for ADC-Net training to achieve robust dispersion compensation in OCT.

Functional Optical Coherence Tomography for Intrinsic Signal Optoretinography: Recent Developments and Deployment Challenges.

Intrinsic optical signal (IOS) imaging of the retina, also termed as optoretinogram or optoretinography (ORG), promises a non-invasive method for the objective assessment of retinal function. By providing the unparalleled capability to differentiate individual retinal layers, functional optical coherence tomography (OCT) has been actively investigated for intrinsic signal ORG measurements. However, clinical deployment of functional OCT for quantitative ORG is still challenging due to the lack of a standardized imaging protocol and the complication of IOS sources and mechanisms. This article aims to summarize recent developments of functional OCT for ORG measurement, OCT intensity- and phase-based IOS processing. Technical challenges and perspectives of quantitative IOS analysis and ORG interpretations are discussed.

Depth-resolved vascular profile features for artery-vein classification in OCT and OCT angiography of human retina.

This study is to characterize reflectance profiles of retinal blood vessels in optical coherence tomography (OCT), and to test the potential of using these vascular features to guide artery-vein classification in OCT angiography (OCTA) of the human retina. Depth-resolved OCT reveals unique features of retinal arteries and veins. Retinal arteries show hyper-reflective boundaries at both upper (inner side towards the vitreous) and lower (outer side towards the choroid) walls. In contrast, retinal veins reveal hyper-reflectivity at the upper boundary only. Uniform lumen intensity was observed in both small and large arteries. However, the venous lumen intensity was dependent on the vessel size. Small veins exhibit a hyper-reflective zone at the bottom half of the lumen, while large veins show a hypo-reflective zone at the bottom half of the lumen.

Functional optoretinography: concurrent OCT monitoring of intrinsic signal amplitude and phase dynamics in human photoreceptors.

Intrinsic optical signal (IOS) imaging promises a noninvasive method for objective assessment of retinal function. This study demonstrates concurrent optical coherence tomography (OCT) of amplitude-IOS and phase-IOS changes in human photoreceptors. A new procedure for differential-phase-mapping (DPM) is validated to enable depth-resolved phase-IOS imaging. Dynamic OCT revealed rapid amplitude-IOS and phase-IOS changes, which occur almost right away after the stimulus onset. These IOS changes were predominantly observed within the photoreceptor outer segment (OS), particularly two boundaries connecting to the inner segment and retinal pigment epithelium. The comparative analysis supports that both amplitude-IOS and phase-IOS attribute to transient OS morphological change associated with phototransduction activation in retinal photoreceptors. A simulation modeling is proposed to discuss the relationship between the photoreceptor OS length and phase-IOS changes.

In vivo optoretinography of phototransduction activation and energy metabolism in retinal photoreceptors.

The objective of this study is to verify the anatomic correlate of the second (2nd) outer retina band in optical coherence tomography (OCT), and to demonstrate the potential of using intrinsic optical signal (IOS) imaging for concurrent optoretinography (ORG) of phototransduction activation and energy metabolism in stimulus activated retinal photoreceptors. A custom-designed OCT was employed for depth-resolved IOS imaging in mouse retina activated by a visible light flicker stimulation. The spatiotemporal properties of the IOS changes at the photoreceptor outer segment (OS) and inner segment (IS) were quantitatively evaluated. Rapid IOS change was observed at the OS almost right away, and the IOS at the IS was relatively slow. Comparative analysis indicates that the OS-IOS reflects transient OS deformation caused by the phototransduction activation, and IS-IOS might reflect the energy metabolism caused by mitochondria activation in retinal photoreceptors. The consistency of the distribution of the IS-IOS and the 2nd OCT band supports the IS ellipsoid (ISe), which has abundant mitochondria, as the signal source of the 2nd OCT band of the outer retina.

Functional intrinsic optical signal imaging for objective optoretinography of human photoreceptors.

Functional mapping of photoreceptor physiology is important for better disease diagnosis and treatment assessment. Fast intrinsic optical signal (IOS), which arises before light-evoked pupillary response, promises a unique biomarker of photoreceptor physiology for objective optoretinography with high resolution. This study is to test the feasibility of non-mydriatic IOS mapping of retinal photoreceptors in awake human. Depth-resolved optical coherence tomography verified outer segment (OS) as the anatomic origin of fast photoreceptor-IOS. Dynamic IOS changes are primarily confined at OS boundaries connected with inner segment and retinal pigment epithelium, supporting transient OS shrinkage due to phototransduction process as the mechanism of the fast photoreceptor-IOS response.

Piriformospora indica-induced phytohormone changes and root colonization strategies are highly host-specific.

Piriformospora indica, an endophytic fungus of Sebacinales, has a wide host range and promotes the performance of mono- and eudicot plants. Here, we compare the interaction of P. indica with the roots of seven host plants (Anthurium andraeanum, Arabidopsis thaliana, Brassica campestris, Lycopersicon esculentum, Oncidium orchid, Oryza sativa, and Zea mays). Microscopical analyses showed that the colonization time and the mode of hyphal invasion into the roots differ in the symbiotic interactions. Substantial differences between the species were also observed for the levels and accumulation of jasmonate (JA) and gibberellin (GA) and the transcript levels for genes involved in their syntheses. No obvious correlation could be detected between the endogenous JA and/or GA levels and the time point of root colonization in a given plant species. Our results suggest that root colonization strategies and changes in the two phytohormone levels are highly host-specific.

Identification and characterization of PsDREB2 promoter involved in tissue-specific expression and abiotic stress response from Paeonia suffruticosa.

Dehydration-responsive element-binding factor 2 (DREB2) belongs to the C-repeat-binding factor (CBF)/DREB subfamily of proteins. In this study, a 2,245 bp PsDREB2 promoter fragment was isolated from the genome of Paeonia suffruticosa. The fragment was rich in A/T bases and contained TATA box sequences, abscisic acid (ABA)-response elements, and other cis-elements, such as MYB and CAAT box. The promoter was fused with the ß-glucuronidase (GUS) reporter gene to generate an expression vector. Arabidopsis thaliana was transformed with a flower dipping method. Gus activity in different tissues and organs of transgenic plants was determined via histochemical staining and quantified via GUS fluorescence. The activity of promoter regulatory elements in transgenic plants under drought, low-temperature, high-salt, and ABA stresses was analyzed. The results showed that the PsDREB2 gene promoter was expressed in the roots, stems, leaves, flowers, and silique pods but not in the seeds of transgenic Arabidopsis. Furthermore, the promoter was induced by drought, low temperature, high salt, and ABA. Hence, the PsDREB2 promoter is tissue- and stress-specific and can be used in the genetic engineering of novel peony cultivars in the future.

Ectopic expression of the AaFUL1 gene identified in Anthurium andraeanum affected floral organ development and seed fertility in tobacco.

Anthurium andraeanum is a high-grade potted flower that enjoys global popularity. Its floral organs have been substantially modified, and its ornamental value is based on its petaloid bracts. MADS-box gene products are important transcription factors that control plant development. In particular, the APETALA1 (AP1)/FRUITFULL (FUL) family of MADS-box genes plays a key role in flowering transitions and out-whorl floral organ identity specification. In this report, one FUL-like gene was cloned from Anthurium andraeanum and named AaFUL1 after bioinformatics identification. Subsequent subcellular localization experiments confirmed that the AaFUL1 protein was located in the nucleus, and data obtained from an expression analysis indicated that the relative expression level of AaFUL1 was the highest in bracts and inflorescences, while its expression was relatively low in stems and roots. Next, an AaFUL1 overexpression vector was constructed and ectopically expressed in tobacco. The transformants did not show any early flowering phenotype, but the average internode length of the inflorescence branch was significantly higher than that observed in the control, and its petal color had substantially faded. The morphology of the petal and pistil was clearly changed, the fruit was deformed, and the seed was largely aborted. These data indicate that even though the sequence of AaFUL1 is relatively conserved, its function differs from that of other orthologs, and the FUL subfamily of MADS-box transcription factors may have taken on new functions during the evolution processes. The results of this experiment enrich our knowledge of FUL transcription factors in monocotyledon plants.

Overexpression of particular MADS-box transcription factors in heat-stressed plants induces chloroplast biogenesis in petals.

Chloroplasts convert solar energy into biologically useful forms of energy by performing photosynthesis. Although light and particular genes are known to promote chloroplast development, little is known about the mechanisms that regulate the tissue-specificity and cell-specificity of chloroplast biogenesis. Thus, the mechanisms that determine whether non-photosynthetic plastids rather than chloroplasts develop in petals remain largely unexplored. Although heat stress is known to inhibit photosynthesis, we do not know whether heat stress affects chloroplast biogenesis. Here, we report that heat stress up-regulates the expression of chlorophyll biosynthesis-related genes and promotes chloroplasts biogenesis in petals overexpressing SOC1 (suppressor of overexpression of CO) and novel SOC1-like genes. We also found that these specific MADS-box transcription factors are present in most photosynthetic eukaryotes and that the expression of more than one homolog is observed in chloroplast-containing tissues. These findings not only provide novel insights into the tissue specificity of chloroplast biogenesis and a method for producing green petals but also are consistent with heat stress influencing chloroplast biogenesis in higher plants.

Protein responses in kenaf plants exposed to drought conditions determined using iTRAQ technology.

The molecular mechanisms that underlie drought stress responses in kenaf, an important crop for the production of natural fibers, are poorly understood. To address this issue, we describe here the first iTRAQ-based comparative proteomic analysis of kenaf seedlings. Plants were divided into the following three treatment groups: Group A, watered normally (control); Group B, not watered for 6 days (drought treatment); and Group C, not watered for 5 days and then rewatered for 1 day (recovery treatment). A total of 5014 proteins were detected, including 4932 (i.e., 98.36%) that were matched to known proteins in a BLAST search. We detected 218, 107, and 348 proteins that were upregulated in Group B compared with Group A, Group C compared with Group A, and Group B compared with Group C, respectively. Additionally, 306, 145, and 231 downregulated proteins were detected during the same comparisons. Seventy differentially expressed proteins were analyzed and classified into 10 categories: photosynthesis, sulfur metabolism, amino sugar and nucleotide sugar metabolism, oxidative phosphorylation, ribosome, fatty acid elongation, thiamine metabolism, tryptophan metabolism, plant-pathogen interaction, and propanoate. Kenaf adapted to stress mainly by improving the metabolism of ATP, regulating photosynthesis according to light intensity, promoting the synthesis of osmoregulators, strengthening ion transport signal transmission, and promoting metabolism and cell stability. This is the first study to examine changes in protein expression in kenaf plants exposed to drought stress. Our results identified key drought-responsive genes and proteins and may provide useful genetic information for improving kenaf stress resistance.

Edge Defect Engineering of Nitrogen-Doped Carbon for Oxygen Electrocatalysts in Zn-Air Batteries.

Metal-free bifunctional oxygen electrocatalysts are extremely critical to the advanced energy conversion devices, such as high energy metal-air batteries. Effective tuning of edge defects and electronic density on carbon materials via simple methods is especially attractive. In this work, a facile alkali activation method has been proposed to prepare carbon with large specific surface area and optimized porosity. In addition, subsequent nitrogen-doping leads to high pyridinic-N and graphitic-N contents and abundant edge defects, further enhancing electrochemical activities. Theoretical modeling via first-principles calculations has been conducted to correlate the electrocatalytic activities with their fundamental chemical structure of N doping and edge defect engineering. The metal-free product (NKCNPs-900) shows a high half-wave potential of 0.79 V (ORR). Furthermore, the assembled Zn-air batteries display excellent performance among carbon-based metal-free oxygen electrocatalysts, such as large peak power density up to 131.4 mW cm-2, energy density as high as 889.0 W h kg-1 at 4.5 mA cm-2, and remarkable discharge-charge cycles up to 575 times. Preliminarily, the rechargeable nonaqueous Li-air batteries were also investigated. Therefore, our work provides a low-cost, metal-free, and high-performance bifunctional carbon-based electrocatalyst for metal-air batteries.