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.

In silico analysis of the wild-type and mutant-type of BRCA2 gene.

BACKGROUND: The aim of this study was to conduct an in silico analysis of a novel compound heterozygous variant in breast cancer susceptibility gene 2 (BRCA2) to clarify its structure-function relationship and elucidate the molecular mechanisms underlying triple-negative breast cancer (TNBC). METHODS: A tumor biopsy sample was obtained from a 42-year-old Chinese woman during surgery, and a maxBRCA™ test was conducted using the patient's whole blood. We obtained an experimentally determined 3D structure (1mje.pdb) of the BRCA2 protein from the Protein Data Bank (PDB) as a relatively reliable reference. Subsequently, the wild-type and mutant structures were predicted using SWISS-MODEL and AlphaFold, and the accuracy of these predictions was assessed through the SAVES online server. Furthermore, we utilized a high ambiguity-driven protein-protein docking (HADDOCK) algorithm and protein-ligand interaction profiler (PLIP) to predict the pathogenicity of the mutations and elucidate pathogenic mechanisms that potentially underlies TNBC. RESULTS: Histological examination revealed that the tumor biopsy sample exhibited classical pathological characteristics of TNBC. Furthermore, the maxBRCA™ test revealed two compound heterozygous BRCA2 gene mutations (c.7670 C > T.pA2557V and c.8356G > A.pA2786T). Through performing in silico structural analyses and constructing of 3D models of the mutants, we established that the mutant amino acids valine and threonine were located in the helical domain and oligonucleotide binding 1 (OB1), regions that interact with DSS1. CONCLUSION: Our analysis revealed that substituting valine and threonine in the helical domain region alters the structure and function of BRCA2 proteins. This mutation potentially affects the binding of proteins and DNA fragments and disrupts interactions between the helical domain region and OB1 with DSS1, potentially leading to the development of TNBC. Our findings suggest that the identified compound heterozygous mutation contributes to the clinical presentation of TNBC, providing new insights into the pathogenesis of TNBC and the influence of compound heterozygous mutations in BRCA2.

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.

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.

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.

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.

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.

Comparation between Guidance for Judicial Expertise of Medical Malpractice and Medical Association Identification Rules of Medical Damage.

On the basis of retaining the technical identification system of medical negligence, the Medical Association Identification Rules of Medical Damage mainly provides technical services for various types of conciliation work about doctor-patient dispute. Its identification work is still influenced by the thinking of medical negligence technical identification and has certain administrative color. Guidance for Judicial Expertise of Medical Malpractice is mainly reflected that the trial of civil cases and pre-trial mediation of courts need service. Its procedures and evidence review are strictly required by the litigation rules and has the characteristics of public legal services provided as a third-party neutral institution. Technical identification of medical damage, whether organized by the Medical Association or the forensic identification institutions, is carried out under the background of the current Regulations on the Prevention and Treatment of Medical Disputes and the Civil Code of the People's Republic of China; both have a corresponding positive role in regulating the medical damage identification activities, and have also laid a certain foundation for the establishment of a unified identification system in the future in China. To understand the different characteristics of the medical damage identification rules issued by the Chinese Medical Association and the Ministry of Justice, and to improve the understanding of the standardization of the forensic identification of medical damage, a comparative study was conducted on Medical Association Identification Rules of Medical Damage and Guidance for Judicial Expertise of Medical Malpractice from seven aspects: Concept and legal status, entrust of identification, identification acceptance, identification procedures, identification presentation meeting, theory of medical malpractice evaluation, consequences and causality of medical damage. The subject of evaluation, the function of evidence review, the role of consulting experts, the technical standard system of malpractice evaluation and other contents were emphatically analyzed.

Precise 3D computer-generated holography based on non-convex optimization with spherical aberration compensation (SAC-NOVO) for two-photon optogenetics.

Computer-generated holography (CGH) has been adopted in two-photon optogenetics as a promising technique for selective excitation of neural ensembles. However, 3D CGH by nonconvex optimization, the state of art method, is susceptible to imprecise axial positioning, due to the quadratic phase approximation in 3D target generation. Even though the misplacement of targets in conventional CGH can be solved by pre-calibration, it still suffers from low efficiency and poor axial resolution of two-photon excitation. Here, we propose a novel CGH method based on non-convex optimization with spherical aberration compensation (SAC-NOVO). Through numerical simulations and two-photon excitation experiments, we verify that SAC-NOVO could achieve precise axial positioning for single and multiple expanded disk patterns, while ensuring high two-photon excitation efficiency. Besides, we experimentally show that SAC-NOVO enables the suppression of dark target areas. This work shows the superiority of SAC-NOVO for two-photon optogenetics.

In vitro and In vivo digestion comparison of bee pollen with or without wall-disruption.

BACKGROUND: Bee pollen is considered as a treasure trove of human and animal nutrients as a result of its extensive nutritional and therapeutic properties. However, the sophisticated pollen wall can largely limit the digestibility and bioavailability of these nutrients. RESULTS: An ultrasonication and high shear technique was used to break the walls of five species of bee pollen, including rape bee pollen, lotus bee pollen, camellia bee pollen, wuweizi bee pollen and apricot bee pollen. We compared the digestibilities of bee pollen with or without wall-disruption. After in vitro and in vivo digestion, unbroken bee pollen grains were still intact and the fragments of wall-disrupted bee pollen still remained as fragments. Mouse in vivo digestion results suggested that the wall-disrupted bee pollen was more easily emptied from the gastrointestinal tract than unbroken bee pollen. After dynamic in vitro digestion, the digestibilities of protein and crude fat in wall-disrupted bee pollen significantly increased to more than 80%; similarly, the release rates of amino acids and reducing sugars in all wall-disrupted samples were almost 1.5 and 2 times as much as those of unbroken samples. CONCLUSION: Based on the results obtained in the present study, we strongly recommend that bee pollen should be wall-disrupted. © 2020 Society of Chemical Industry.

Large-field-of-view imaging by multi-pupil adaptive optics.

Adaptive optics can correct for optical aberrations. We developed multi-pupil adaptive optics (MPAO), which enables simultaneous wavefront correction over a field of view of 450 × 450 µm2 and expands the correction area to nine times that of conventional methods. MPAO's ability to perform spatially independent wavefront control further enables 3D nonplanar imaging. We applied MPAO to in vivo structural and functional imaging in the mouse brain.

Conformal convolutional neural network (CCNN) for single-shot sensorless wavefront sensing.

Wavefront sensing technique is essential in deep tissue imaging, which guides spatial light modulator to compensate wavefront distortion for better imaging quality. Recently, convolutional neural network (CNN) based sensorless wavefront sensing methods have achieved remarkable speed advantages via single-shot measurement methodology. However, the low efficiency of convolutional filters dealing with circular point-spread-function (PSF) features makes them less accurate. In this paper, we propose a conformal convolutional neural network (CCNN) that boosts the performance by pre-processing circular features into rectangular ones through conformal mapping. The proposed conformal mapping reduces the number of convolutional filters that need to describe a circular feature, thus enables the neural network to recognize PSF features more efficiently. We demonstrate our CCNN could improve the wavefront sensing accuracy over 15% compared to a traditional CNN through simulations and validate the accuracy improvement in experiments. The improved performances make the proposed method promising in high-speed deep tissue imaging.

Adaptive optimization for axial multi-foci generation in multiphoton microscopy.

To improve imaging speed, multifocal excitation is widely adopted as a parallel strategy in laser-scanning microscopy. Specifically, axial multifocal microscopy is popular in neuroscience as it enables functional imaging of neurons in multiple depths simultaneously. However, previous phase searching algorithms for axial multi-foci generation generally generate foci of uniform intensities, which cannot compensate the scattering-induced power loss in deep tissue and causes inhomogeneous excitation. Here, we propose a novel adaptive optimization-based phase-searching method (AdaPS) to generate axial multi-foci with arbitrary intensity modulations for scattering-induced loss compensation. By adopting Adaptive Moment Estimation (Adam) as the searching algorithm, our method could escape from unsatisfactory local minima and stably converge to the optimal phase pattern with errors at least an order of magnitude lower. We validate AdaPS through both numerical simulations and experiments and demonstrate that AdaPS could provide uniform multi-depth imaging in scattering phantom and enable high-fidelity multi-depth recordings of neural network dynamics in mouse brain in vivo.

Overcoming tissue scattering in wide-field two-photon imaging by extended detection and computational reconstruction.

Compared to point-scanning multiphoton microscopy, line-scanning temporal focusing microscopy (LTFM) is competitive in high imaging speed while maintaining tight axial confinement. However, considering its wide-field detection mode, LTFM suffers from shallow penetration depth as a result of the crosstalk induced by tissue scattering. In contrast to the spatial filtering based on confocal slit detection, here we propose the extended detection LTFM (ED-LTFM), the first wide-field two-photon imaging technique to extract signals from scattered photons and thus effectively extend the imaging depth. By recording a succession of line-shape excited signals in 2D and reconstructing signals under Hessian regularization, we can push the depth limitation of wide-field imaging in scattering tissues. We validate the concept with numerical simulations, and demonstrate the performance of enhanced imaging depth in in vivo imaging of mouse brains.

Stretchable and upconversion-luminescent polymeric optical sensor for wearable multifunctional sensing.

Stretchable sensors with multiple sensory functions are in high demand for healthcare monitoring and artificial intelligence. Despite recent advances in wearable electronic sensors, it remains a significant challenge to achieve simultaneous sensing of both thermal and mechanical stimuli with a single sensor while integrating high stretchability. Herein, a stretchable and multifunctional optical sensor (SMOS) with simultaneous readout of temperature and strain is developed for wearable physiological monitoring of the human body. The SMOS primarily consists of a stretchable optical sensing fiber made from polymer nanocomposites containing lanthanide-based upconversion nanoparticles (UCNPs). Temperature measurements are achieved by ratiometric intensity measurements of the dual-emission UCNPs upon near-infrared excitation. By virtue of the ratiometric detection, the temperature readout of the SMOS is independent of strain deformations, enabling stable and continuous measurements of skin temperature during body motions. Furthermore, deformation of the SMOS by stretching leads to detectable and reversible changes in its light transmission, allowing tensile strains to be simultaneously measured. As a proof of concept, we demonstrate the capabilities of the SMOS in real time and simultaneous detection of both skin temperature and motion activities of the human body.

Soft and Stretchable Polymeric Optical Waveguide-Based Sensors for Wearable and Biomedical Applications.

The past decades have witnessed the rapid development in soft, stretchable, and biocompatible devices for applications in biomedical monitoring, personal healthcare, and human-machine interfaces. In particular, the design of soft devices in optics has attracted tremendous interests attributed to their distinct advantages such as inherent electrical safety, high stability in long-term operation, potential to be miniaturized, and free of electromagnetic interferences. As the alternatives to conventional rigid optical waveguides, considerable efforts have been made to develop light-guiding devices by using various transparent and elastic polymers, which offer desired physiomechanical properties and enable wearable/implantable applications in optical sensing, diagnostics, and therapy. Here, we review recent progress in soft and stretchable optical waveguides and sensors, including advanced structural design, fabrication strategies, and functionalities. Furthermore, the potential applications of those optical devices for various wearable and biomedical applications are discussed. It is expected that the newly emerged soft and stretchable optical technologies will provide a safe and reliable alternative to next-generation, smart wearables and healthcare devices.

Quantum Dots-Doped Tapered Hydrogel Waveguide for Ratiometric Sensing of Metal Ions.

Advances in fluorescent nanomaterials and photonics have led to a new generation of photonic devices for applications in biosensing, diagnostics, and therapy. However, for clinical utility, biocompatibility and limited light guiding in tissues pose significant challenges. Here, we report a new type of soft, biocompatible, and tapered optical waveguide with capability of delivering light in deep tissues and demonstrate it as a ratiometric probe for rapid point-of-care detection of metal ions. The waveguide was made from quantum dots (QDs)-incorporated biocompatible hydrogels and coated with a thin sensing film to ensure fast exchanges with the surrounding analytes. The tapered design of the waveguide allows more light extraction for efficient excitation of the coating film. To achieve ratiometric measurements, two types of QDs with well-resolved emission bands are synthesized and immobilized in the waveguide and the coating film, respectively. We show that the ratiometric readout of the waveguide sensor is free of environmental disturbances and exhibits negligible drifts when applied in various environments such as being immersed in water or embedded in tissues. The waveguide device provides a new photonic-sensing platform that may allow being engineered to sense a wide range of metal ions and analytes.

Continuous volumetric imaging via an optical phase-locked ultrasound lens.

In vivo imaging at high spatiotemporal resolution is key to the understanding of complex biological systems. We integrated an optical phase-locked ultrasound lens into a two-photon fluorescence microscope and achieved microsecond-scale axial scanning, thus enabling volumetric imaging at tens of hertz. We applied this system to multicolor volumetric imaging of processes sensitive to motion artifacts, including calcium dynamics in behaving mouse brain and transient morphology changes and trafficking of immune cells.

Enhancing axial resolution and background rejection in line-scanning temporal focusing microscopy by focal modulation.

Compared with two-photon point-scanning microscopy, line-scanning temporal focusing microscopy breaks the limitation on imaging rate and maintains the axial resolution, which makes it promising for various biomedical studies. However, for deep tissue imaging, it suffers from reduced axial resolution and increased background noise due to sample induced wavefront distortion. Here, we propose a spatio-spectral focal modulation technique to enhance axial resolution and background rejection by simply subtracting an aberrated image, which is induced by a spatial light modulator, from an unaberrated image. The proposed technique could improve the axial resolution by a factor of 1.3 in our implementation, verified by both simulations and experiments. Besides, we show that compared with spatial modulation alone, spatio-spectral modulation induces less peak intensity loss caused by image subtraction. We further demonstrate the performance of our technique on the enhanced axial resolution and background rejection by deep imaging of cleared mouse brains and in vivo imaging of living mouse brains.

Multiplexed static FBG strain sensors by dual-comb spectroscopy with a free running fiber laser.

We demonstrate a novel and compact FBG interrogation system for multiplexed static strain sensing with a free running mode-locked fiber laser. Multiplexed FBG array in cascading are interrogated by coherent dual-comb pulses generated from a single fiber laser. Dual-comb spectroscopy is achieved with the fiber laser to precisely detect the strain-induced spectral shifts of the FBG sensors. Multi-point strain measurements are performed to characterize the proposed system, where a large dynamic range of 520 µÎµ with 0.5 µÎµ resolution is achieved.