A ratiometric SERS strategy for the prediction of cancer cell proportion and guidance of glioma surgical resection.

Rapid and accurate identification of tumor boundaries is critical for the cure of glioma, but it is difficult due to the invasive nature of glioma cells. This paper aimed to explore a rapid diagnostic strategy based on a label-free surface-enhanced Raman scattering (SERS) technique for the quantitative detection of glioma cell proportion intraoperatively. With silver nanoparticles as substrate, an in-depth SERS analysis was performed on simulated clinical samples containing normal brain tissue and different concentrations of patient-derived glioma cells. The results revealed two universal characteristic peaks of 655 and 717 cm-1, which strongly correlated with glioma cell proportion regardless of individual differences. Based on the intensity ratio of the two peaks, a ratiometric SERS strategy for the quantification of glioma cells was established by employing an artificial neuron network model and a polynomial regression model. Such a strategy accurately estimated the proportion of glioma cells in simulated clinical samples (R2 = 0.98) and frozen samples (R2 = 0.85). More importantly, it accurately facilitated the delineation of tumor margins in freshly obtained samples. Taken together, this SERS-based method ensured a rapid and more detailed identification of tumor margins during surgical resection, which could be beneficial for intraoperative decision-making and pathological evaluation.

Convolutional sparse coding for compressed sensing photoacoustic CT reconstruction with partially known support.

In photoacoustic tomography (PAT), imaging speed is an essential metric that is restricted by the pulse laser repetition rate and the number of channels on the data acquisition card (DAQ). Reconstructing the initial sound pressure distribution with fewer elements can significantly reduce hardware costs and back-end acquisition pressure. However, undersampling will result in artefacts in the photoacoustic image, degrading its quality. Dictionary learning (DL) has been utilised for various image reconstruction techniques, but they disregard the uniformity of pixels in overlapping blocks. Therefore, we propose a compressive sensing (CS) reconstruction algorithm for circular array PAT based on gradient domain convolutional sparse coding (CSCGR). A small number of non-zero signal positions in the sparsely encoded feature map are used as partially known support (PKS) in the reconstruction procedure. The CS-CSCGR-PKS-based reconstruction algorithm can use fewer ultrasound transducers for signal acquisition while maintaining image fidelity. We demonstrated the effectiveness of this algorithm in sparse imaging through imaging experiments on the mouse torso, brain, and human fingers. Reducing the number of array elements while ensuring imaging quality effectively reduces equipment hardware costs and improves imaging speed.

Trait-based approach revealed the seasonal variation of mesozooplankton functional groups in the South Yellow Sea.

Functional traits determine the fitness of organisms and mirror their ecological functions. Although trait-based approaches provide ecological insights, it is underexploited for marine zooplankton, particularly with respect to seasonal variation. Here, based on four major functional traits, including body length, feeding type, trophic group, and reproduction mode, we quantified the seasonal variations of mesozooplankton functional groups in the South Yellow Sea (SYS) in the spring, summer, and autumn of 2018. Strong seasonal dynamics were identified for all traits but patterns varied among traits. Small zooplankton (47.7-88.6%), omnivores-herbivores (81.3-97.6%), and free spawners (54.8-92.5%) dominated in three seasons, while ambush feeders and current feeders dominated in spring (45.7%), and autumn (73.4%), respectively. Cluster analysis of the functional traits showed that the mesozooplankton in the SYS can be classified into eight functional groups. The biogeographic and seasonal variations of functional groups can be partially explained by environmental drivers. Group 1, represented by omnivores-herbivores, was the most dominant functional group, the abundance of which peaked in spring and was positively correlated with chlorophyll a concentration, indicating its close association with phytoplankton dynamics. The contribution of giant, active ambush carnivores, passive ambush carnivore jellyfish, current omnivores-detritivores, and parthenogenetic cladocerans increased with sea surface temperature. The proportion of giant, active ambush carnivores and active ambush omnivore-carnivore copepods decreased with salinity in autumn. This study presents a new perspective for understanding the dynamics of zooplankton and paves the way for further research on the functional diversity of zooplankton in the SYS. Supplementary Information: The online version contains supplementary material available at 10.1007/s42995-022-00156-9.

Latitudinal transition of epipelagic mesozooplankton in the northwestern Pacific in winter.

The northwestern Pacific (NWP) is a hotspot of marine biodiversity study, and zooplankton is a crucial secondary producer in the marine ecosystem. It is of utmost importance to do extensive study on the distribution of zooplankton community in the NWP. The distribution of epipelagic zooplankton community in the 143-146°E section between the equator and 36°N in winter was examined in this study. The findings indicated that the Kuroshio extension, the North Pacific Subtropical Gyre, the North Equatorial Current and the North Equatorial Countercurrent were the four main ocean currents in the NWP that regulated the latitudinal transition of epipelagic mesozooplankton and split the sample section into four station groups. The key factors influencing zooplankton's geographic distribution were temperature, primary productivity, and current movement. In general, as latitude increased, zooplankton abundance and biomass first decreased and subsequently flourished in the NWP. Diversity indexes and indicator species also revealed the difference across the communities in four station groups. The body length of zooplankton declined gradually from north to south under the combined influence of temperature and primary production, with the minimum in the oligotrophic subtropical zone and a sub-peak in the high primary production area near the equator. Additionally, the abundance of zooplankton was higher in the winter than in the summer due to seasonal fluctuations in the properties of the ocean currents. This study elucidated the control effects of ocean currents on the latitudinal distribution of zooplankton, supplemented records of the geographic distribution and body length characteristics of zooplankton communities in the NWP, and provided the basis for further research on the ecological role of zooplankton communities and the global changes of marine ecosystems.

Reduced Fitness and Elevated Oxidative Stress in the Marine Copepod Tigriopus japonicus Exposed to the Toxic Dinoflagellate Karenia mikimotoi.

Blooms of the toxic dinoflagellate Karenia mikimotoi cause devastation to marine life, including declines of fitness and population recruitment. However, little is known about the effects of them on benthic copepods. Here, we assessed the acute and chronic effects of K. mikimotoi on the marine benthic copepod Tigriopus japonicus. Results showed that adult females maintained high survival (>85%) throughout 14-d incubation, but time-dependent reduction of survival was detected in the highest K. mikimotoi concentration, and nauplii and copepodites were more vulnerable compared to adults. Ingestion of K. mikimotoi depressed the grazing of copepods but significantly induced the generation of reactive oxygen species (ROS), total antioxidant capacity, activities of antioxidant enzymes (superoxide dismutase, catalase, and glutathione peroxidase), and acetylcholinesterase. Under sublethal concentrations for two generations, K. mikimotoi reduced the fitness of copepods by prolonging development time and decreasing successful development rate, egg production, and the number of clutches. Our findings suggest that the bloom of K. mikimotoi may threaten copepod population recruitment, and its adverse effects are associated with oxidative stress.

Diversity, prevalence, and expression of cyanase genes (cynS) in planktonic marine microorganisms.

Cyanate is utilized by many microbes as an organic nitrogen source. The key enzyme for cyanate metabolism is cyanase, converting cyanate to ammonium and carbon dioxide. Although the cyanase gene cynS has been identified in many species, the diversity, prevalence, and expression of cynS in marine microbial communities remains poorly understood. Here, based on the full-length cDNA sequence of a dinoflagellate cynS and 260 homologs across the tree of life, we extend the conserved nature of cyanases by the identification of additional ultra-conserved residues as part of the modeled holoenzyme structure. Our phylogenetic analysis showed that horizontal gene transfer of cynS appears to be more prominent than previously reported for bacteria, archaea, chlorophytes, and metazoans. Quantitative analyses of marine planktonic metagenomes revealed that cynS is as prevalent as ureC (urease subunit alpha), suggesting that cyanate plays an important role in nitrogen metabolism of marine microbes. Highly abundant cynS transcripts from phytoplankton and nitrite-oxidizing bacteria identified in global ocean metatranscriptomes indicate that cyanases potentially occupy a key position in the marine nitrogen cycle by facilitating photosynthetic assimilation of organic N and its remineralisation to NO3 by the activity of nitrifying bacteria.

Effects of Prorocentrum donghaiense bloom on zooplankton functional groups in the coastal waters of the East China Sea.

Blooms of the non-toxic dinoflagellate Prorocentrum donghaiense are common in the East China Sea; however, the in situ impacts of these blooms on zooplankton community functions have not yet been conducted in this area. Using functional trait-based methods, we found that P. donghaiense bloom significantly changed the zooplankton community structure and functions in the coastal water of the East China Sea. Zooplankton species richness and biodiversity increased after the bloom. Based on body length, feeding type, trophic group and reproductive mode, we categorized zooplankton into four functional groups and characterized their dynamics. Before and after the bloom, the zooplankton community was dominated by herbivorous- and free-spawner- lineages represented by copepods and tunicates, while during the bloom, the zooplankton community was dominated by carnivorous- and egg-brooding- lineages represented by small jellyfish, chaetognaths and copepods.

Measurement of Iris Thickness at Different Regions in Healthy Chinese Adults.

PURPOSE: To study the variation of iris thicknesses in different regions and explore the possible correlations with age and gender. METHODS: Healthy Chinese adults were recruited; the anterior segment of their eyes was imaged by swept-source optical coherence tomography (SS-OCT). The horizontal scan of the right eye was selected, and the thicknesses of both the nasal and temporal irises were measured at 199 evenly spaced points. RESULTS: A total of 233 subjects with an average age of 36.79 ± 10.04 years (range 19 to 62) were included in the final analysis. The mean iris thicknesses of the temporal and nasal sides were 364.79 ± 47.58 µm and 372.44 ± 43.75 µm, respectively. The mean nasal iris thickness was positively correlated with age (ß = 0.9 µm/year; P = 0.002), but the temporal one was not (ß = 0.077 µm/year; P = 0.806). At 139 points of the nasal iris and 146 points of the temporal iris, the iris thickness was significantly correlated with age (P < 0.05). The thicknesses of the peripheral and pupillary parts were positively correlated with age, while the middle part was negatively correlated with age. No significant difference was observed in the mean iris thickness between genders (temporal: t = 1.597, P = 0.112; nasal: t = 1.870, P = 0.063), but females had a thicker iris than males at 50 points in the temporal side and 49 points in the nasal side (P < 0.05); no point in males was observed to have thicker iris compared to females. CONCLUSION: Using SS-OCT and the novel method, thicknesses of the iris at different regions were measured. The thicknesses of the peripheral and pupillary irises increase with age, while the thicknesses of the middle part decrease.

CXCL1 stimulates decidual angiogenesis via the VEGF-A pathway during the first trimester of pregnancy.

Angiogenesis is critical to establishing a successful pregnancy. The chemokine (C-X-C motif) ligand 1 (CXCL1) is a small cytokine belonging to the CXC chemokine family that is an important chemokine involved in the processes of angiogenesis and arteriogenesis; however, little is known about its role in decidual angiogenesis. Effects of CXCL1 on cell proliferation and migration (propidium iodide staining and wound healing assays) of HUVEC cells were determined. The angiogenesis roles of CXCL1 in HUVEC-HTR8/SVneo co-culture system were detected by the tube formation assay. Signal transduction pathways in HUVEC cells in response to CXCL1 were determined by in-cell western analyses. In vivo, mice were injected with (1) PBS (Group A) or (2) CXCL1-neutralizing antibody (Group B) or (3) CXCL1-neutralizing antibody plus recombinant VEGF-A protein (Group C) from E1 to E5 and sacrificed at E6.5 of pregnancy. The decidual angiogenesis in mice was examined by immunohistochemistry of cluster designation 34 (CD34), and the expression levels of vascular endothelial growth factor-A (VEGF-A) in the decidual cells and vascular endothelial growth factor receptor 2 (VEGFR2) in decidual vascular endothelial cells were also tested. Exogenous recombinant human CXCL1 supported endothelial cell proliferation and migration, and this effect was blocked by CXCL1-neutralizing antibody or CXCR2 inhibitor SB265610. The tube formation of HUVEC-HTR8/SVneo co-culture system was significantly stimulated by CXCL1, but this effect was markedly abrogated once they were pretreated with CXCL1-neutralizing antibody or CXCR2 inhibitor SB265610. In addition, the level of vascular endothelial growth factor A (VEGF-A) expression in HUVEC cells was increased by CXCL1, and this level was suppressed by CXCL1-neutralizing antibody or CXCR2 inhibitor SB265610. In vivo, compared with Group A (n = 3), decidual angiogenesis was significantly reduced in Group B by CD34 immunostaining. But compared with Group B, decidual angiogenesis was significantly increased in Group C. In addition, the expression of VEGF-A and VEGFR2 was significantly increased after neutralizing of CXCL1 in Group B. In conclusions, CXCL1 may play essential roles in decidual angiogenesis during the first trimester, and this function may be mediated in part via altering VEGF-A expression.

Automatic Classification of Anterior Chamber Angle Based on Ultrasound Biomicroscopy Images.

INTRODUCTION: Evaluating the anterior chamber angle (ACA) is important for the early diagnosis and treatment of primary angle-closure glaucoma. The assessment of ultrasound biomicroscopy (UBM) images usually requires well-trained ophthalmologists and screening for patients with narrow ACA is usually time- and labor-intensive. Therefore, the automatic assessment of UBM could be cost-effective and valuable in daily practice. OBJECTIVE: The objective of this study is to develop an automatic method for localizing and classifying ACA based on UBM images. METHODS: UBM images were collected and a coarse-to-fine method was used to localize the apex of the angle recess. By analyzing the grayscale features around the angle recess, closed angles were identified, and the rest were then classified as open or narrow angles, based on the degree of ACA. Using manual classification as the reference standard, the overall accuracy (OAcc), sensitivity (Sen), specificity (Spe), and balanced accuracy of the automatic classification method were evaluated. RESULTS: A total of 540 UBM images from 290 participants were analyzed. Using these UBM images and the proposed method, the ACA was classified as open, narrow, or closed. During processing, the method localized the angle recess with 95% accuracy. The OAcc of the ACA classification was 77.8%, and the Spe and Sen of our method were 85.8 and 81.7% for angle closure; 88.9 and 75.6% for open angles; 91.9 and 76.1% for narrow angles, respectively. CONCLUSIONS: Our method of automatic angle localization and classification based on UBM images is feasible and reliable. The automatic classification of ACA provides a basis and reference for future studies.

Interplane bulk motion analysis and removal based on normalized cross-correlation in optical coherence tomography angiography.

Bulk motion seriously degrades the image quality of optical coherence tomography angiography (OCTA). Conventional correction methods focus on in-plane displacement, while the bulk motion component perpendicular to B-scans also introduces noise. This work first presents an evaluation of this component using a specific scan protocol and an approximate expression derived from peak-normalized cross-correlation values, and then quantitatively assesses how interplane bulk motion noise reduce the sensitivity of cross-sectional angiograms. Finally, we developed a repetitive bulk motion correction method based on the estimated displacements and redundant volume scans. The correction does not require registration and angiogram reconstruction of low flow sensitivity frames, and the results of in vivo mice skin OCTA imaging experiments show that the proposed method can effectively reduce bulk motion noise caused by cardiac and respiratory motion and occasional shaking, and improve OCTA image quality, which has practical significance for clinical OCTA diagnosis and analysis.

Quality improvement of adaptive optics retinal images using conditional adversarial networks.

The adaptive optics (AO) technique is widely used to compensate for ocular aberrations and improve imaging resolution. However, when affected by intraocular scatter, speckle noise, and other factors, the quality of the retinal image will be degraded. To effectively improve the image quality without increasing the imaging system's complexity, the post-processing method of image deblurring is adopted. In this study, we proposed a conditional adversarial network-based method for directly learning an end-to-end mapping between blurry and restored AO retinal images. The proposed model was validated on synthetically generated AO retinal images and real retinal images. The restoration results of synthetic images were evaluated with the metrics of peak signal-to-noise ratio (PSNR), structural similarity (SSIM), perceptual distance, and error rate of cone counting. Moreover, the blind image quality index (BIQI) was used as the no-reference image quality assessment (NR-IQA) algorithm to evaluate the restoration results on real AO retinal images. The experimental results indicate that the images restored by the proposed method have sharper quality and higher signal-to-noise ratio (SNR) when compared with other state-of-the-art methods, which has great practical significance for clinical research and analysis.

Ratiometric fluorescence method for ctDNA analysis based on the construction of a DNA four-way junction.

We present a novel ratiometric fluorescent biosensor for ctDNA analysis based on the construction of a DNA four-way junction (FWJ). Three fuel strands for the FWJ are firstly designed and prepared. Another essential strand for the formation of the structure is the DNA product generated from target ctDNA initiated strand displacement amplification. With the transformation of the DNA structure, the FRET states of two fluorophores change and the ratiometric fluorescence response can be recorded to indicate the level of the initial ctDNA. The proposed method also has excellent capability to discriminate mismatches and shows potential practical utility for clinical samples.

Bright carbon nanodots for miRNA diagnostics coupled with concatenated hybridization chain reaction.

We developed a bright carbon nanodot-based miRNA detection method with signal amplification by concatenated hybridization chain reaction (CHCR). In the presence of target miRNA, CHCR was triggered and a frond-like DNA product was formed, which recovered remarkable fluorescence. The location and level of the target miRNA were then indicated.

Feasibility of microscope-integrated swept-source optical coherence tomography in canaloplasty.

BACKGROUND: Several researchers have used commercial microscope-integrated optical coherence tomography (OCT) systems in glaucoma surgery, including ab interno trabeculectomy and canaloplasty. However, the 840 nm wavelength light source of the OCT systems is not ideal for imaging the anterior chamber angle structures because of its limited penetration. We evaluated the potential value of a microscope-integrated swept-source OCT system with a 1,310 nm center-wavelength light in canaloplasty for glaucoma. METHODS: Sixteen porcine eyes were used to simulate canaloplasty. The critical surgical steps were monitored using a prototype microscope-integrated OCT system with a 1,310 nm light source and a high axial scan rate of 100 kHz. The images from swept-source OCT and three-dimensional images from the microscope were projected simultaneously onto a liquid crystal display three-dimensional monitor (LMD-4251TD, Sony, Japan). The changes in the collector vessel (aqueous drainage structure in the porcine eye, similar to Schlemm's canal in humans) were measured using Image J software. Histological sections stained with hematoxylin and eosin were used to assess surgical efficacy. RESULTS: High-resolution real-time images of the anterior segment were acquired during canaloplasty using the microscope-integrated OCT system. With the real-time OCT images, the position of the collector vessel was identified and the scleral flap could be created at the ideal location. The expansion of the collector vessel after viscoelastic injection was also visualized in real time. Compared with baseline, there was a significant increase in the cross-sectional area (from 14,502.98±9,242.55 to 59,499.04±20,506.41 µm2, P<0.001) of the collector vessel. CONCLUSIONS: Using the microscope-integrated OCT system, real-time images of the anterior segment were successfully acquired during the operation. The microscope-integrated OCT system might be useful in future anti-glaucoma surgery.

Self-protective transcriptional alterations in ZF4 cells exposed to Pb(NO3 )2 and AgNO3.

In this study, gene expression alterations of phase I to III enzymes, transcription factors, and microRNA (miRNA) in embryonic zebrafish fibroblasts (ZF4) cells after the treatment of Pb(NO3 )2 and AgNO3 were investigated, to illustrate the possible detoxification pathway of heavy metal ions. It was observed that both metals caused concentration-dependent death and moderate elevation of oxidative stress in ZF4 cells. In response to such toxicity, upregulation of multidrug resistance protein (mdr)4 and multiresistance-associated protein (mrp)1 were found. However, enhanced expression of glutathione S-transferase (gst) and cytochrome P450 (cyp)1a could only be detected during the exposure of Pb2+ . In addition, both metals induced extensive upregulation of pregnane X receptor (pxr), but only moderate elevation of E2-related factor (nrf2), while they suppressed the expression of miR-122 and miR-126. In conclusion, Pb2+ and Ag+ shared the same detoxification mechanism including ABC transporters, Pxr, and miRNA in ZF4 cells, which needs further investigation.

Automatic Classification of Anterior Chamber Angle Using Ultrasound Biomicroscopy and Deep Learning.

PURPOSE: To develop a software package for automated classification of anterior chamber angle of the eye by using ultrasound biomicroscopy. METHODS: Ultrasound biomicroscopy images were collected, and the trabecular-iris angle was manually measured and classified into three categories: open angle, narrow angle, and angle closure. Inception v3 was used as the classifying convolutional neural network and the algorithm was trained. RESULTS: With a recall rate of 97% in the test set, the neural network's classification accuracy can reach 97.2% and the overall area under the curve was 0.988. The sensitivity and specificity were 98.04% and 99.09% for the open angle, 96.30% and 98.13% for the narrow angle, and 98.21% and 99.05% for the angle closure categories, respectively. CONCLUSIONS: Preliminary results show that an automated classification of the anterior chamber angle achieved satisfying sensitivity and specificity and could be helpful in clinical practice. TRANSLATIONAL RELEVANCE: The present work suggests that the algorithm described here could be useful in the categorizing of anterior chamber angle and screening for subjects who are at high risk of angle closure.

Altered Gene expression of ABC transporters, nuclear receptors and oxidative stress signaling in zebrafish embryos exposed to CdTe quantum dots.

Adenosine triphosphate-binding cassette (ABC) transporters, including P-glycoprotein (Pgp) and multi-resistance associated proteins (Mrps), have been considered important participants in the self-protection of zebrafish embryos against environmental pollutants, but their possible involvement in the efflux and detoxification of quantum dots (QDs), as well as their regulation mechanism are currently unclear. In this work, gene expression alterations of ABC transporters, nuclear receptors, and oxidative stress signaling in zebrafish embryos after the treatment of mercaptopropionic acid (MPA)CdTe QDs and MPA-CdSCdTe QDs were investigated. It was observed that both QDs caused concentration-dependent delayed hatching effects and the subsequent induction of transporters like mrp1&2 in zebrafish embryos, indicating the protective role of corresponding proteins against CdTe QDs. Accompanying these alterations, expressions of nuclear receptors including the pregnane X receptor (pxr), aryl hydrocarbon receptor (ahr) 1b, and peroxisome proliferator-activated receptor (ppar)-ß were induced by QDs in a concentration- and time-dependent manner. Moreover, elevated oxidative stress, reflected by the reduction of glutathione (GSH) level and superoxide dismutase (SOD) activities, as well as the dramatic induction of nuclear factor E2 related factor (nrf) 2, was also found. More importantly, alterations of pxr and nrf2 were more pronounced than that of mrps, and these receptors exhibited an excellent correlation with delayed hatching rate in the same embryos (R2 > 0.8). Results from this analysis demonstrated that the induction of mrp1 and mrp2 could be important components for the detoxification of QDs in zebrafish embryos. These transporters could be modulated by nuclear receptors and oxidative stress signaling. In addition, up-regulation of pxr and nrf2 could be developed as toxic biomarkers of CdTe QDs.

Ultrasensitive Detection of DNA Based on Exonuclease III-Assisted Recycling Amplification and DNAzyme Motor.

Herein, we have developed a dual amplification strategy for ultrasensitive detection of DNA combining exonuclease III (Exo III)-assisted reaction and DNAzyme motor. DNA probes are carefully designed; thus, target recognition and the first amplification cycle are accomplished simultaneously, which makes the operation very convenient. Moreover, the self-powered DNAzyme motor may translate a single binding event into cleavage of multiple fluorescence probes, which significantly heightens the signal intensity. As a result, the limit of detection as low as 21 fM is achieved. The fluorescence intensity is found to have a linear relationship with respect to the logarithm of DNA concentration in a wide range from 100 fM to 10 nM. This proposed method shows great potential for the applications of biological studies and clinical diagnosis.