Carbene Footprinting Directs Design of Genetically Encoded Proximity-Reactive Protein Binders.

Genetically encoding proximal-reactive unnatural amino acids (PrUaas), such as fluorosulfate-l-tyrosine (FSY), into natural proteins of interest (POI) confer the POI with the ability to covalently bind to its interacting proteins (IPs). The PrUaa-incorporated POIs hold promise for blocking undesirable POI-IP interactions. Selecting appropriate PrUaa anchor sites is crucial, but it remains challenging with the current methodology, which heavily relies on crystallography to identify the proximal residues between the POIs and the IPs for the PrUaa anchorage. To address the challenge, here, we propose a footprinting-directed genetically encoded covalent binder (footprinting-GECB) approach. This approach employs carbene footprinting, a structural mass spectrometry (MS) technique that quantifies the extent of labeling of the POI following the addition of its IP, and thus identifies the responsive residues. By genetically encoding PrUaa into these responsive sites, POI variants with covalent bonding ability to its IP can be produced without the need for crystallography. Using the POI-IP model, KRAS/RAF1, we showed that engineering FSY at the footprint-assigned KRAS residue resulted in a KRAS variant that can bind irreversibly to RAF1. Additionally, we inserted FSY at the responsive residue in RAF1 upon footprinting the oncogenic KRASG12D/RAF1, which lacks crystal structure, and generated a covalent binder to KRASG12D. Together, we demonstrated that by adopting carbene footprinting to direct PrUaa anchorage, we can greatly expand the opportunities for designing covalent protein binders for PPIs without relying on crystallography. This holds promise for creating effective PPI inhibitors and supports both fundamental research and biotherapeutics development.

A New Denoising Method for Belt Conveyor Roller Fault Signals.

In the process of the intelligent inspection of belt conveyor systems, due to problems such as its long duration, the large number of rollers, and the complex working environment, fault diagnosis by acoustic signals is easily affected by signal coupling interference, which poses a great challenge to selecting denoising methods of signal preprocessing. This paper proposes a novel wavelet threshold denoising algorithm by integrating a new biparameter and trisegment threshold function. Firstly, we elaborate on the mutual influence and optimization process of two adjustment parameters and three wavelet coefficient processing intervals in the BT-WTD (the biparameter and trisegment of wavelet threshold denoising, BT-WTD) denoising model. Subsequently, the advantages of the proposed threshold function are theoretically demonstrated. Finally, the BT-WTD algorithm is applied to denoise the simulation signals and the vibration and acoustic signals collected from the belt conveyor experimental platform. The experimental results indicate that this method's denoising effectiveness surpasses that of traditional threshold function denoising algorithms, effectively addressing the denoising preprocessing of idler roller fault signals under strong noise backgrounds while preserving useful signal features and avoiding signal distortion problems. This research lays the theoretical foundation for the non-contact intelligent fault diagnosis of future inspection robots based on acoustic signals.

Super-Selective Intra-arterial Thrombolytic Therapy for Peripheral Facial Paralysis After Postauricular Hyaluronic Acid Injection for the Esthetic Correction of Lying Ears.

BACKGROUND: Hyaluronic acid (HA) injection in the auricular base is one of the most popular and non-surgical cosmetic procedures for correcting lying ears and optimizing the facial profile because of its minimal invasiveness, immediate effect and safety (Li et al. in Aesthet Surg J 44: 746-75, 2024). But we have recently discovered that this treatment may lead to a new and rare complication called peripheral facial paralysis that has never been reported before. Until now, the etiology, clinical traits, treatment strategies, outcomes and possible reversibility have not been characterized. METHODS: In the present study, we enrolled 4 patients with peripheral facial paralysis after subcutaneous postauricular HA filler injection. Preoperative digital subtraction angiography revealed a vascular embolism. Then, the patients underwent super-selective facial arterial thrombolytic therapy via hyaluronidase and papaverine injections. Simultaneously, general symptomatic treatment and nutritional therapy were performed. RESULTS: The patients were relieved of their clinical symptoms and the significant improvement was observed in terms of motor function in her left facial areas after treatment. The auricular skin necrosis of all patients was restored to near normal appearance. CONCLUSION: Our results indicate that super-selective facial arterial thrombolytic therapy is feasible for patients with peripheral facial paralysis induced by HA embolism. It was also beneficial in the recovery from skin necrosis. The therapy was shown to be worthy of clinical application. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Novel Techniques in Microscopy: introduction to the feature issue.

The editors introduce the feature issue on "Novel Techniques in Microscopy," which was the topic of a symposium held on April 24-27, 2023, in Vancouver, BC. This symposium was part of the Optics in the Life Sciences Congress.

Challenges and solutions for the downstream purification of therapeutic proteins.

The innovation in recombinant protein technology has brought forth a host of challenges related to the purification of these therapeutic proteins. This article delves into the intricate landscape of developing purification processes for artificially designed therapeutic proteins. The key hurdles include controlling protein reduction, protein capture, ensuring stability, eliminating aggregates, removing host cell proteins and optimizing protein recovery. In this review, we outline the purification strategies in order to obtain products of high purity, highlighting the corresponding solutions to circumvent the unique challenges presented by recombinant therapeutic proteins, and exemplify the practical applications by case studies. Finally, a perspective towards future purification process development is provided.

Surface Modification of Nano-Hydroxyapatite/Polymer Composite for Bone Tissue Repair Applications: A Review.

Nano-hydroxyapatite (n-HA) is the main inorganic component of natural bone, which has been widely used as a reinforcing filler for polymers in bone materials, and it can promote cell adhesion, proliferation, and differentiation. It can also produce interactions between cells and material surfaces through selective protein adsorption and has therefore always been a research hotspot in orthopedic materials. However, n-HA nano-particles are inherently easy to agglomerate and difficult to disperse evenly in the polymer. In addition, there are differences in trace elements between n-HA nano-particles and biological apatite, so the biological activity needs to be improved, and the slow degradation in vivo, which has seriously hindered the application of n-HA in bone fields, is unacceptable. Therefore, the modification of n-HA has been extensively reported in the literature. This article reviewed the physical modification and various chemical modification methods of n-HA in recent years, as well as their modification effects. In particular, various chemical modification methods and their modification effects were reviewed in detail. Finally, a summary and suggestions for the modification of n-HA were proposed, which would provide significant reference for achieving high-performance n-HA in biomedical applications.

Label-free multimodal imaging with simultaneous two-photon and three-photon microscopy and kernel-based nonlinear scaling denoising.

We report on a compact multimodal imaging system that can acquire two-photon microscopy (2PM) and three-photon microscopy (3PM) images simultaneously. With dual excitation wavelengths, multiple contrasts including two-photon-excitation-fluorescence (2PEF), second harmonic generation (SHG), and third harmonic generation (THG) are acquired simultaneously from cells, collagen fibers, and interfaces, all label-free. Challenges related to the excitation by two wavelengths and the effective separation of 2PM and 3PM signals are discussed and addressed. The data processing challenge where multiple contrasts can have significantly varying signal levels is also addressed. A kernel-based nonlinear scaling (KNS) denoising method is introduced to reduce noise from ultra-low signal images and generate high-quality multimodal images. Simultaneous 2PM and 3PM imaging is demonstrated on various tissue samples. The simultaneous acquisition speeds up the imaging process and minimizes the commonly encountered problem of motion artifacts and mechanical drift in sequential acquisition. Multimodal imaging with simultaneous 2PM and 3PM will have great potential for label-free in-vivo imaging of biological tissues.

Carbon capture and utilization by algae with high concentration CO2 or bicarbonate as carbon source.

Algae plays a key role in carbon capture and utilization (CCU) as it can capture and use the atmospheric CO2 for conversion of value-added products. Concentrated CO2 is common in flue gas and provides opportunities for algae cultivation. The drawbacks are mass transfer limitation, poor CO2 dissolution, and challenges to reach optimal levels for algal growth at given flue gas levels. Bicarbonate is flexible to be used as carbon source and owns the potential to enhance the efficiency of biological carbon fixation by algae. The requirements of algae strains are more stringent. To improve the industrial scale-up of CCU, system optimization is of great importance. More novel algal strains that can grow rapidly under harsh environment and provide valuable bio-products should be developed for large-scale production. Algae-driven CCU is promising for achieving carbon-neutrality.

W-Net: A boundary-aware cascade network for robust and accurate optic disc segmentation.

Accurate optic disc (OD) segmentation has a great significance for computer-aided diagnosis of different types of eye diseases. Due to differences in image acquisition equipment and acquisition methods, the resolution, size, contrast, and clarity of images from different datasets show significant differences, resulting in poor generalization performance of deep learning networks. To solve this problem, this study proposes a multi-level segmentation network. The network includes data quality enhancement module (DQEM), coarse segmentation module (CSM), localization module (OLM), and fine segmentation stage module (FSM). In FSM, W-Net is proposed for the first time, and boundary loss is introduced in the loss function, which effectively improves the performance of OD segmentation. We generalized the model in the REFUGE test dataset, GAMMA dataset, Drishti-GS1 dataset, and IDRiD dataset, respectively. The results show that our method has the best OD segmentation performance in different datasets compared with state-of-the-art networks.

Effect of H2O2-VC degradation on structural characteristics and immunomodulatory activity of larch arabinogalactan.

The arabinogalactan in the representative softwood biomass of larch was degraded using an environmentally friendly hydrogen peroxide and vitamin C (H2O2-VC) system to improve its immunomodulatory activity. Through the H2O2-VC degradation mechanism, hydroxyl radicals are generated, which then target the hydrogen atoms within polysaccharides, resulting in the breaking of glycosidic bonds. Given the impact of oxidative degradation on polysaccharides, we identified three specific arabinogalactan degradation products distinguished by their arabinosyl side chain compositions. The primary structures of the degradation products were investigated using Fourier-transform infrared spectroscopy and nuclear magnetic resonance spectroscopy. Congo red staining showed that the degradation products were absent in the triple-helix structure. The results of the in vitro immunological experiments indicated that an appropriate reduction in the molar ratio of arabinose to galactose enhanced the immunostimulatory effects on RAW 264.7 cells. In addition, the immunostimulatory pathway mediated by arabinogalactan was explored by toll-like receptor 4 (TLR4) inhibitor (TAK-242) These findings provide novel insights into the understanding of the relationship between the structure of arabinogalactan and its biological activity.