Removing crosstalk signals in neuron activity by time multiplexed excitations in a two-photon all-optical physiology system.

The two-photon all-optical physiology system has attracted great interest in deciphering neuronal circuits in vivo, benefiting from its advantages in recording and modulating neuronal activities at single neuron resolutions. However, the interference, or crosstalk, between the imaging and photostimulation beams introduces a significant challenge and may impede the future application of voltage indicators in two-photon all-optical physiology system. Here, we propose the time multiplexed excitation method to distinguish signals from neuronal activities and crosstalks from photostimulation. In our system, the laser pulses of the imaging beam and photostimulation beam are synchronized, and a time delay is introduced into these pulses to separate the fluorescence signal generated by these two beams. We demonstrate the efficacy of our system in eliminating crosstalk signals from photostimulation and evaluate its influence on both genetically encoded calcium indicators (GECIs) and genetically encoded voltage indicators (GEVIs) through in vivo experiments.

Single-cell profiling transcriptomic reveals cellular heterogeneity and cellular crosstalk in choroidal neovascularization model.

Choroidal neovascularization (CNV) is a hallmark of neovascular age-related macular degeneration (nAMD) and a major contributor to vision loss in nAMD cases. However, the identification of specific cell types associated with nAMD remains challenging. Herein, we performed single-cell sequencing to comprehensively explore the cellular diversity and understand the foundational components of the retinal pigment epithelium (RPE)/choroid complex. We unveiled 10 distinct cell types within the RPE/choroid complex. Notably, we observed significant heterogeneity within endothelial cells (ECs), fibroblasts, and macrophages, underscoring the intricate nature of the cellular composition in the RPE/choroid complex. Within the EC category, four distinct clusters were identified and EC cluster 0 was tightly associated with choroidal neovascularization. We identified five clusters of fibroblasts actively involved in the pathogenesis of nAMD, influencing fibrotic responses, angiogenic effects, and photoreceptor function. Additionally, three clusters of macrophages were identified, suggesting their potential roles in regulating the progression of nAMD through immunomodulation and inflammation regulation. Through CellChat analysis, we constructed a complex cell-cell communication network, revealing the role of EC clusters in interacting with fibroblasts and macrophages in the context of nAMD. These interactions were found to govern angiogenic effects, fibrotic responses, and inflammatory processes. In summary, this study reveals noteworthy cellular heterogeneity in the RPE/choroid complex and provides valuable insights into the pathogenesis of CNV. These findings will open up potential avenues for deep understanding and targeted therapeutic interventions in nAMD.

Challenges and opportunities in inflammatory bowel disease: from current therapeutic strategies to organoid-based models.

BACKGROUND: Inflammatory bowel disease (IBD) is an increasingly prevalent global health concern that has garnered substantial attention. However, the underlying mechanisms are still unclear and the current treatments have significant limitations. Intestinal organoids provide an in vitro model to explore the pathogenesis, test the therapeutic effects, and develop regenerative treatments as well as offer the potential to transform drug discovery of IBD. METHODS: To advance our understanding of the whole story of IBD spanning from the pathogenesis to the current therapeutic strategies and latest advancements, a comprehensive search of major databases including PubMed, Scopus, and Web of Science was conducted to retrieve original articles and reviews related to IBD, organoids, pathogenesis and therapy. RESULTS: This review deciphers the etiopathogenesis and the current therapeutic approaches in the treatment of IBD. Notably, critical aspects of intestinal organoids in IBD, such as their potential applications, viability, cell renewal ability, and barrier functionality are highlighted. We also discuss the advances, limitations, and prospects of intestinal organoids for precision medicine. CONCLUSION: The latest strides made in research about intestinal organoids help elucidate intricate aspects of IBD pathogenesis, and pave the prospective avenues for novel therapeutic interventions.

Transfer RNA derived fragment, tRF-Glu-CTC, aggravates the development of neovascular age-related macular degeneration.

Rationale: Angiogenesis expedites tissue impairment in many diseases, including age-related macular degeneration (AMD), a leading cause of irreversible blindness in elderly. A substantial proportion of neovascular AMD patients, characterized by aberrant choroidal neovascularization (CNV), exhibit poor responses or adverse reactions to anti-VEGF therapy. Herein, we aimed to unveil the function of newly identified transfer RNA-derived small RNA, tRF-Glu-CTC, in the pathology of CNV and determine its potential in inhibiting angiogenesis. Methods: Small non-coding RNA sequencing and quantitative polymerase chain reaction were conducted to detect expression pattern of tRF-Glu-CTC in CNV development. Immunofluorescence staining, fundus fluorescein angiography and ex vivo choroidal sprouting assays were employed for the evaluation of tRF-Glu-CTC's function in CNV development. The role of tRF-Glu-CTC in endothelial cells were determined by in vitro endothelial cell proliferation, migration and tube formation assays. Transcriptome sequencing, dual-luciferase reporter assay and in vitro experiments were conducted to investigate downstream mechanism of tRF-Glu-CTC mediated pathology. Results: tRF-Glu-CTC exhibited substantial up-regulation in AMD patients, laser-induced CNV model, and endothelial cells under hypoxia condition, which is a hallmark of CNV. Inhibiting tRF-Glu-CTC reduced angiogenesis and hypoxia stress in the neovascular region without neuroretina toxicity in laser-induced CNV model, showing an anti-angiogenic effect comparable to bevacizumab, while overexpression of tRF-Glu-CTC significantly augmented CNV. Mechanically, under hypoxia condition, angiogenin was involved in the production of tRF-Glu-CTC, which in turn triggered endothelial cell tubulogenesis, migration and promoted the secretion of inflammatory factors via the suppression of vasohibin 1 (VASH1). When downregulating VASH1 expression, the inhibition of tRF-Glu-CTC showed minimal suppression on angiogenesis. Conclusions: This study demonstrated the important role of tRF-Glu-CTC in the progression of angiogenesis. Targeting of tRF-Glu-CTC may be an alternative to current anti-VEGF therapy for CNV in AMD and other conditions with angiogenesis.

Nitrogen addition does not alter symmetric responses of soil respiration to changing precipitation in a semi-arid grassland.

Concurrent changing precipitation regimes and atmospheric nitrogen (N) deposition can have profound influences on soil carbon (C) cycling. However, how N enrichment regulates the responses of soil C fluxes to increasing variability of precipitation remains elusive. As part of a field precipitation gradient experiment with nine levels of precipitation amounts (-60 %, -45 %, -30 %, -15 %, ambient precipitation, +15 %, +30 %, +45 %, and +60 %) and two levels of N addition (0 and 10 g N m-2 yr-1) in a semi-arid temperate steppe on the Mongolian Plateau, this work was conducted to investigate the responses of soil respiration to decreased and increased precipitation (DP and IP), N addition, and their possible interactions. Averaged over the three years from 2019 to 2021, DP suppressed soil respiration by 16.1 %, whereas IP stimulated it by 27.4 %. Nitrogen addition decreased soil respiration by 7.1 % primarily via reducing microbial biomass C. Soil respiration showed symmetric responses to DP and IP within all the four precipitation variabilities (i.e., 15 %, 30 %, 45 %, and 60 %) under ambient N. Nevertheless, N addition did not alter the symmetric responses of soil respiration to changing precipitation due to the comparable sensitivities of microbial biomass and root growth to DP and IP under the N addition treatment. These findings indicate that intensified precipitation variability does not change but N addition could alleviate soil C releases. The unchanged symmetric responses of soil respiration to precipitation variability under N addition imply that N deposition may not change the response pattern of soil C releases to predicted increases in precipitation variability in grasslands, facilitating the robust projections of ecosystem C cycling under future global change scenarios.

Insights into the complex interactions between Rab22a and extracellular vesicles in cancers.

INTRODUCTION: Oncogenic Ras-related GTP-binding proteins, referred to as Rabs, are characterized by their intricate interactions with upstream, downstream molecules, and notably, extracellular vesicles (EVs). While the expansive family of Rabs and their associated signaling pathways have been exhaustively dissected, Rab22a emerges as an entity of outstanding interest, owing to its potent influence in many biological processes and its conspicuous correlation with cancer metastasis and migration. A burgeoning interest in the interactions between Rab22a and EVs in the field of oncology underscores the necessity for more in-depth reviews and scholarly discourses. METHODS: We performed a review based on published original and review articles related to Rab22a, tumor, microRNA, exosome, microvesicles, EVs, CD147, lysosome, degradation, endosomal recycling, etc. from PubMed, Web of Science and Google Scholar databases. RESULTS AND CONCLUSIONS: We summarize the regulatory processes governing the expression of Rab22a and the mutants of Rab22a. Notably, the present understanding of complex interactions between Rab22a and EVs are highlighted, encompassing both the impact of Rab22a on the genesis of EVs and the role of EVs that are affected by Rab22a mutants in propelling tumor advancement. The dynamic interaction between Rab22a and EVs plays a significant role in the progression of tumors, and it can provide novel insights into the pathogenesis of cancers and the development of new therapeutic targets.

Zinc protoporphyrin IX generation by Leuconostoc strains isolated from bulged pasteurized vacuum sliced hams.

The colour of meat typically fades as it decays. However, it has been observed that certain vacuum-packaged spoiled hams can maintain a pink colour even when the packaging is bulged. A large amount of Zinc protoporphyrin IX (ZnPP) was found in these hams, compared to fresh red hams or spoiled and grey hams. Combined with high-throughput sequencing and cultural isolation, the potential cultures of Leuconostoc mesenteroides S-13 (LM), Leuconostoc citreum OCLC11 (LC), and Leuconostoc mesenteroides subsp. IMAU:80679 (LS) were selected based on their ability to produce ZnPP. Subsequently, these cultures were introduced into a fermented sausage model to assess their effect on colour conversion. The analysis of absorption and fluorescent spectra showed that Nitrite sausages contained nitrosyl heme pigment, while bacteria-inoculated sausages were predominantly composed of ZnPP. In addition, the a* value of the LS sausage was close to the Nitrite group at the end of fermentation, significantly higher than control, indicating the effect of bacterial metabolism on the redness. Meanwhile, the Ferrochelatase (FECH) activity of LM, LC and LS groups were 140 ± 13, 113 ± 16 and 201 ± 20 U/g sausage, respectively, providing a potential method on compensating for nitrite/nitrate substitution based on the presence of ZnPP in meat products.

The dual role of microglia in neuropathic pain after spinal cord injury: Detrimental and protective effects.

Spinal cord injury (SCI) is a debilitating condition that is frequently accompanied by neuropathic pain, resulting in significant physical and psychological harm to a vast number of individuals globally. Despite the high prevalence of neuropathic pain following SCI, the precise underlying mechanism remains incompletely understood. Microglia are a type of innate immune cell that are present in the central nervous system (CNS). They have been observed to have a significant impact on neuropathic pain following SCI. This article presents a comprehensive overview of recent advances in understanding the role of microglia in the development of neuropathic pain following SCI. Specifically, the article delves into the detrimental and protective effects of microglia on neuropathic pain following SCI, as well as the mechanisms underlying their interconversion. Furthermore, the article provides a thorough overview of potential avenues for future research in this area.

Optimizing e-commerce warehousing through open dimension management in a three-dimensional bin packing system.

In the field of e-commerce warehousing, maximizing the utilization of packing bins is a fundamental goal for all major logistics enterprises. However, determining the appropriate size of packing bins poses a practical challenge for many logistics companies. Given the limited research on the open-size 3D bin packing problem as well as the high complexity and lengthy computation time of existing models, this study focuses on optimizing multiple-bin sizes within the e-commerce context. Building upon existing research, we propose a hybrid integer programming model, denoted as the three dimensional multiple option dimensional rectangular packing problem (3D-MODRPP), to address the multiple-bin size 3D bin packing problem. Additionally, we leverage well-established hardware and software technologies to propose a 3D bin packing system capable of accommodating multiple bin types with open dimensions. To reduce the complexity of the model and the number of constraints, we introduce a novel assumption method for 0-1 integer variables in the overlap and rotation constraints. By applying this approach, we significantly streamline the computational complexity associated with the model calculations. Furthermore, we refine the dataset by developing a customized version based on the classical Three-Dimensional One-Size Dependent Rectangular Packing Problem (3D-ODRPP) dataset, leading to improved outcomes. Through comprehensive analysis of the research results, our model exhibits remarkable advancements in addressing the strong heterogeneous bin packing problem, the weak heterogeneous bin packing problem, the actual bin packing problem, and the bin packing problem with multiple bin types and open sizes. Specifically, it significantly reduces model complexity and computation time and increases space utilization. The system designed in this study paves the way for practical applications based on the proposed model, providing researchers with broader research prospects and directions to expand the scope of investigation in the field of 3D bin packing. Consequently, this system contributes to solving complex 3D packing problems, reducing space waste, and enhancing transportation efficiency.

Upconversion-Luminescent Fiber Microchannel Sensors for Temperature Monitoring at High Spatial Resolution in the Brains of Freely Moving Animals.

Brain temperature is a critical factor affecting neural activity and function, whose fluctuations may result in acute life-threatening health complications and chronic neuropathology. To monitor brain temperature, luminescent nanothermometry (LN) based on upconversion nanoparticles (UCNPs) with low autofluorescence has received extensive attention for its advantages in high temperature sensitivity and high response speed. However, most of current the LNs are based on optical imaging, which fails in temperature monitoring in deep brain regions at high spatial resolution. Here, the fiber microchannel sensor (FMS) loaded with UCNPs (UCNP-FMS) is presented for temperature monitoring at high spatial resolution in the deep brains of freely moving animals. The UCNP-FMS is fabricated by incorporating UCNPs in microchannels of optical fibers, whose diameter is ∼50 µm processed by femtosecond laser micromachining for spatially resolved sensing. The UCNPs provide thermal-sensitive upconversion emissions at dual wavelengths for ratiometric temperature sensing, ensuring a detection accuracy of ± 0.3 °C at 37 °C. Superior performances of UCNP-FMS are demonstrated by real-time temperature monitoring in different brain regions of freely moving animals under various conditions such as taking food, undergoing anesthesia/wakefulness, and suffering external temperature changes. Moreover, this study shows the capability of UCNP-FMS in distributed temperature sensing in mammalian brains in vivo.

Computational Study of Drop-on-Demand Coaxial Electrohydrodynamic Jet and Printing Microdroplets.

Currently, coaxial electrohydrodynamic jet (CE-Jet) printing is used as a promising technique for the alternative fabrication of drop-on-demand micro- and nanoscale structures without using a template. Therefore, this paper presents numerical simulation of the DoD CE-Jet process based on a phase field model. Titanium lead zirconate (PZT) and silicone oil were used to verify the numerical simulation and the experiments. The optimized working parameters (i.e., inner liquid flow velocity 150 m/s, pulse voltage 8.0 kV, external fluid velocity 250 m/s, print height 16 cm) were used to control the stability of the CE-Jet, avoiding the bulging effect during experimental study. Consequently, different sized microdroplets with a minimum diameter of ~5.5 µm were directly printed after the removal of the outer solution. The model is considered the easiest to implement and is powerful for the application of flexible printed electronics in advanced manufacturing technology.

The mechanism of Leuconostoc mesenteroides subsp. IMAU:80679 in improving meat color: Myoglobin oxidation inhibition and myoglobin derivatives formation based on multi enzyme-like activities.

The Leuconostoc mesenteroides subsp. IMAU:80679 (LM) was chosen for its superior capability in enhancing redness, and was incubated in a broth system containing metmyoglobin (MetMb) to investigate its mechanisms for color improvement. The a* value of LM group reached its highest level of 52.75 ± 1.04 at 24 h, significantly higher than control of 19.75 ± 0.6 (p < 0.05). The addition of LM could inhibit myoglobin oxidation to some extent. Meanwhile, higher content of nitrosylmyoglobin (NOMb) and Zn-protoporphyrin (Znpp) were observed in LM samples during the whole incubation period. Furthermore, enzymatic activity and encoded genes related to MetMb reduction and pigment formation were determined to explain its possible mechanism on color enhancement. Finally, by extracting crude enzymes and adding them to meat batters, the redness of crude enzyme group was comparable to that achieved with 20 ppm nitrite, providing a potential method on compensating for nitrite/nitrate substitution in meat products.

Comprehensive metabolic profiling of diabetic retinopathy.

Diabetic retinopathy (DR) is an important complication of diabetes mellitus and a prevalent blind-causing ophthalmic disease. Despite years of efforts, rapid and accurate diagnosis of DR remains a challenging task. Metabolomics has been used as a diagnostic tool for disease progression and therapy monitoring. In this study, retinal tissues were collected from diabetic mice and age-matched non-diabetic mice. An unbiased metabolic profiling was performed to identify the altered metabolites and metabolic pathways in DR. 311 differential metabolites were identified between diabetic retinas and non-diabetic retinas under the criteria of variable importance in projection (VIP) > 1 and P < 0.05. These differential metabolites were highly enriched in purine metabolism, amino acid metabolism, glycerophospholipid metabolism, and pantaothenate and CoA biosynthesis. We then evaluated the sensitivity and specificity of purine metabolites as the candidate biomarkers for DR through the area under the receiver-operating characteristic curves (AUC-ROCs). Compared with other purine metabolites, adenosine, guanine, and inosine had higher sensitivity, specificity, and accuracy for DR prediction. In conclusion, this study sheds new light on the metabolic mechanism of DR, which can facilitate clinical diagnosis, therapy, and prognosis of DR in the future.

Plug-and-play fiber-optic sensors based on engineered cells for neurochemical monitoring at high specificity in freely moving animals.

In vivo detection of neurochemicals, including neurotransmitters and neuromodulators, is critical for both understanding brain mechanisms and diagnosing brain diseases. However, few sensors are competent in monitoring neurochemical dynamics in vivo at high specificity. Here, we propose the fiber-optic probes based on engineered cells (FOPECs) for plug-and-play, real-time detection of neurochemicals in freely moving animals. Taking advantages of life-evolved neurochemical receptors as key components, the chemical specificity of FOPECs is unprecedented. We demonstrate the applications of FOPECs in real-time monitoring of neurochemical dynamics under various physiology and pathology conditions. With no requirement of viral infection in advance and no dependence on animal species, FOPECs can be widely adopted in vertebrates, such as mice, rats, rabbits, and chickens. Moreover, FOPECs can be used to monitor drug metabolisms in vivo. We demonstrated the neurochemical monitoring in blood circulation systems in vivo. We expect that FOPECs will benefit not only neuroscience study but also drug discovery.

Botanical Origins and Antioxidant Activities of Two Types of Flavonoid-Rich Poplar-Type Propolis.

(1) Background: Propolis has attracted attention in recent years due to its important pharmacological effects. The present study aimed to investigate the botanical origins of 39 propolis samples and evaluate their antioxidant activities; (2) Methods: A HPLC-PDA system was used to analyze the phenolic compositions of propolis and poplar bud resin samples. The antioxidant activities of propolis samples were evaluated by oxygen radical absorption capacity (ORAC) and superoxide anion free radical scavenging capacity assay; (3) Results: Our study shows that 17 propolis samples were characterized by five predominant flavonoids, including 5-methoxy pinobanksin, pinobanksin, pinocembrin, pinobanksin-3-acetate, and chrysin, while 22 propolis samples were characterized by four flavonoids (pinobanksin, pinocembrin, pinobanksin-3-acetate, and chrysin). The average contents of characteristic flavonoids reached up to over 70% and 65% of total phenolics, respectively. Furthermore, the botanical origins of the two types of propolis samples were identified as Populus × euramericana cv. 'Neva' and Populus Simonii × P. nigra, respectively; (4) Conclusions: Most notably, our results reveal that these propolis samples presented excellent antioxidant activities due to their high contents of flavonoid. These flavonoid-rich propolis samples can thus be used to develop low-allergen and high-antioxidant nutraceuticals.

Crucial roles of Rab22a in endosomal cargo recycling.

Endosomal cargo recycling lies at the heart of subcellular trafficking processes under the management of several Ras-related GTP-binding proteins (Rabs) which are coordinated by their upstream regulators and require their downstream effectors to display their functions. In this regard, several Rabs have been well-reviewed except Rab22a. Rab22a is a crucial regulator of vesicle trafficking, early endosome and recycling endosome formation. Notably, recent studies demonstrated the immunological roles of Rab22a, which are closely associated with cancers, infection and autoimmune disorders. This review provides an overview of the regulators and effectors of Rab22a. Also, we highlight the current knowledge of the role of Rab22a in endosomal cargo recycling, including the biogenesis of recycling tubules with the help of a complex with Rab22a at its core, and how different internalized cargo chooses different recycling routes thanks to the cooperation of Rab22a, its effectors and its regulators. Of note, contradictions and speculation related to endosomal cargo recycling that Rab22a brings impacts on are also discussed. Finally, this review endeavors to briefly introduce the various events impacted by Rab22a, particularly focusing on the commandeered Rab22a-associated endosomal maturation and endosomal cargo recycling, in addition to the extensively investigated oncogenic role of Rab22a.

Ag-doped Cu nanosheet arrays for efficient hydrogen evolution reaction.

A zinc-infiltration process was adopted to prepare silver-doped copper nanosheet arrays. The larger atomic radius of Ag introduces tensile stress, which lowers the electron density at the s-orbitals of Cu atoms and improves the adsorption capability for hydrogen atoms. As a catalyst for hydrogen evolution, these silver doped copper nanosheet arrays achieved a low overpotential of 103 mV at 10 mA cm-2 in 1 M KOH, which is 604 mV lower than that of pure copper foil.

Evaluating the Genetic Background Effect on Dissecting the Genetic Basis of Kernel Traits in Reciprocal Maize Introgression Lines.

Maize yield is mostly determined by its grain size. Although numerous quantitative trait loci (QTL) have been identified for kernel-related traits, the application of these QTL in breeding programs has been strongly hindered because the populations used for QTL mapping are often different from breeding populations. However, the effect of genetic background on the efficiency of QTL and the accuracy of trait genomic prediction has not been fully studied. Here, we used a set of reciprocal introgression lines (ILs) derived from 417F × 517F to evaluate how genetic background affects the detection of QTLassociated with kernel shape traits. A total of 51 QTL for kernel size were identified by chromosome segment lines (CSL) and genome-wide association studies (GWAS) methods. These were subsequently clustered into 13 common QTL based on their physical position, including 7 genetic-background-independent and 6 genetic-background-dependent QTL, respectively. Additionally, different digenic epistatic marker pairs were identified in the 417F and 517F ILs. Therefore, our results demonstrated that genetic background strongly affected not only the kernel size QTL mapping via CSL and GWAS but also the genomic prediction accuracy and epistatic detection, thereby enhancing our understanding of how genetic background affects the genetic dissection of grain size-related traits.

Compact, Hybrid Light-Sheet and Fourier Light-Field Microscopy with a Single Objective for High-Speed Volumetric Imaging In Vivo.

Volumetric imaging of biodynamics at high spatiotemporal resolutions in vivo is vital in biomedical studies, in which Fourier light field microscopy (FLFM) is a promising technique. However, the commonly used wide-field illumination strategy in FLFM introduces intense out of depth-of-field background, which not only degrades the image quality, but also introduces reconstruction artifacts. Employing light sheet illumination is an effective way to alleviate the background and reduce photobleaching in light-field microscopy. Unfortunately, the introduction of light-sheet illumination often requires an extra objective and precise alignment, which increases the system complexity. Here, we propose the compact, hybrid light-sheet and FLFM (CLS-FLFM), which uses only a single objective to achieve both light-sheet illumination and Fourier light-field imaging simultaneously. With a micromirror under the objective, we focus the light sheet, which ensures selective-volume-illumination, on the imaging plane of the FLFM to perform volumetric imaging. We demonstrate the superior performance of CLS-FLFM in inhibiting background in both structural and dynamical imaging of larval zebrafish in vivo. We envision that CLS-FLFM finds wide applications in high-speed, background-inhibited volumetric imaging of biodynamics in vivo.