Detection of urea in milk by urease-inorganic hybrid nanoflowers combined with portable colorimetric microliter tube.

A simple one-pot green synthesis method was used to prepare urease-inorganic hybrid nanoflowers (UE-HNFs), which had a high surface-to-volume ratio to improve enzyme catalytic efficiency and make urease reusable. A portable colorimetric microliter tube based on urease-inorganic hybrid nanoflowers (UE-HNFs-PCMT), as an urea colorimetric biosensor, was developed for determining urea concentration in milk. The combination of urea colorimetric biosensor and a smartphone is used for capturing the colour change of milk after reaction. There was a good linear relationship between colour intensity of the image (Δ intensity) and urea concentration (43-600 mg L-1), with a detection limit of 12.81 mg L-1. UE-HNFs-PCMT has the advantages of no need for complex equipment, easy operation, reusability, low detection cost, good portability, and environmental friendliness and can achieve urea detection in milk.

Novel three-stage microalgal cultivation system for lipid production utilizing nutrients derived from refinery waste.

The pollution and transformation of refineries are receiving increasing attention. The carbonic anhydrase in Tetradesmus obliquus was found exhibiting a hysteresis phenomenon in response to periodic changes in the composition of external carbon sources, with a surge in inorganic carbon concentration stressing the carbonic anhydrase activity to increase by 6-9 times. On this basis, a novel three-stage culture system of T. obliquus was proposed, which mainly uses refinery waste as the nutrients. By controlling the nutrient content in the environment, especially the composition of carbon sources, microalgae could sequentially complete rapid biomass accumulation, efficient inorganic carbon assimilation, and oil production. Compared to a single-environment culture system, the biomass yield increased by 1.34 times, the oil content increased by more than 6%, and the oil productivity increased by 2.08 times. Above findings may lay a partial theoretical foundation for the future evolution of traditional refineries towards "fossil-algal-biomass" hybrid refineries.

Improved Prognosis in HER2-Low Breast Cancer Patients with Reduced Ki67 Index After Neoadjuvant Chemotherapy: A Multi-Center Retrospective Study.

Background: HER2-low breast cancer represents a distinct subgroup with unique clinical characteristics and treatment challenges. Nevertheless, it remains uncertain whether there exists a distinction in δKi67 between patients with HER2-low and HER2-zero statuses, and whether the prognosis varies among patients with differing HER2 statuses and δKi67. Methods: We conducted a multi-center retrospective study to investigate the correlation between alterations in Ki67 index following NAC and the prognosis among patients with HER2-low or HER2-zero breast cancer. 3 distinct cohorts comprising patients with HER2-negative breast cancer who underwent NAC were included. Comprehensive clinicopathological data were documented, with particular emphasis on evaluating changes in Ki67 index from baseline to post-NAC. These changes were then correlated with disease-free survival (DFS) through rigorous analysis. Results: Three cohorts, comprising 403, 315, and 72 patients respectively, were finally included. The study found that δKi67 did not show significant associations with other variables and were not identified as independent risk factors for survival. Nevertheless, across the three cohorts, following NAC, HER2-low breast cancer patients with δKi67 below the cut-off value demonstrated a better prognosis compared to those with δKi67 above the cut-off value. Additionally, their prognosis was also superior to that of HER2-zero breast cancer patients with δKi67 below the cut-off value. Conclusion: Our study demonstrates that HER2-low breast cancer patients with δKi67 values below the cut-off point after NAC are associated with improved prognosis. Monitoring δKi67 index and HER2 status may help identify patients who are likely to benefit from NAC and guide personalized treatment strategies.

Single-shot 20-fold expansion microscopy.

Expansion microscopy (ExM) is in increasingly widespread use throughout biology because its isotropic physical magnification enables nanoimaging on conventional microscopes. To date, ExM methods either expand specimens to a limited range (~4-10× linearly) or achieve larger expansion factors through iterating the expansion process a second time (~15-20× linearly). Here, we present an ExM protocol that achieves ~20× expansion (yielding <20-nm resolution on a conventional microscope) in a single expansion step, achieving the performance of iterative expansion with the simplicity of a single-shot protocol. This protocol, which we call 20ExM, supports postexpansion staining for brain tissue, which can facilitate biomolecular labeling. 20ExM may find utility in many areas of biological investigation requiring high-resolution imaging.

Implementation of Chief Complaint-Based Clinical Reasoning Training in Periodontal Internships.

BACKGROUND: The gap between theoretical knowledge and clinical skills highlights the need for clinical reasoning training curriculum in periodontal education, especially in periodontal internships. This study aims to develop a Chief Complaint-Based Clinical Reasoning Training (CCB-CRT) program and evaluate its impact on periodontal interns' clinical reasoning abilities and overall performance. METHODS: The CCB-CRT program was developed based on eight common chief complaints (CCs) identified through surveys of periodontal specialists and an analysis of patient visit data from a university-affiliated hospital's periodontal clinic. The study involved a comparison between a control group of fifth-year dental students (2021) and a CCB-CRT group (2022). Both groups completed an 8-week training course. The CCB-CRT group received additional training focused on the 8 common CCs, using student-led discussions, flipped classroom, mind mapping, and presentations. Evaluation criteria included overall performance, disease diagnosis and treatment plan, misdiagnosis rates, and students' satisfaction. RESULTS: After 1 year of CCB-CRT implementation, participants in the CCB-CRT group showed substantial improvements in overall performance, diagnostic accuracy, and satisfaction compared to traditional teaching methods. The program enhanced students' understanding of theoretical knowledge, improved their interpretation of clinical manifestations and examination results, and enhanced their clinical reasoning skills and diagnostic accuracy. CONCLUSIONS: The successful application of the CCB-CRT program in periodontology education demonstrates its efficacy in improving clinical reasoning skills and diagnostic efficiency among students. The structured approach facilitates the transition from theoretical knowledge to practical application, contributing to better patient care in periodontal practice.

Renal-Clearable Organic Probes From D-A-D Type Aza-BODIPY Fluorophores for Multiphoton Deep-Brain Imaging.

Bright near-infrared (NIR) fluorescent probes play an important role in in vivo optical imaging. Here, renal-clearable nanodots prepared from Aza-BODIPY are reported fluorophores for multiphoton brain imaging. The design of donor-acceptor-donor (D-A-D) type conjugated structures endowed the fluorophores with large three-photon absorption cross-section for both 1620 and 2200 nm excitation. The side chain modification and lipid encapsulation yield ultrasmall nanodots (≈4 nm) and a high fluorescence quantum yield (≈0.35) at 720 nm emission in the aqueous phase. The measured three-photon action cross-section of a single Aza-BODIPY fluorophore in the nanodots is ≈30 times higher than the commonly used Sulforhodamine 101 dye. Three-photon deep brain imaging of subcortical structures is demonstrated, reaching a depth of 1900 µm below the brain surface in a live mouse study. The nanodots enabled blood flow measurement at a depth of 1617 µm using line scanning three-photon microscopy (3PM). This work provides superior fluorescent probes for multiphoton deep-brain imaging.

Transforming restored heavy metal-contaminated soil into eco-friendly bricks: An insight into heavy metal stabilization and environmental safety.

Materialization is currently the primary method for utilizing restored heavy metal-contaminated soil (RHMCS). However, compared to ordinary building materials, the migration and transformation mechanisms of heavy metals (HMs) while preparing these materials remain unclear. To bridge these gaps, this study investigated the migration and transformation mechanisms of As and Pb during the sintering of RHMCS into bricks. This study is the first to conduct a systematic study from the perspectives of both the inner and outer brick layers on the patterns and mechanisms of HM migration and transformation during the sintering process, along with the safety of product utilization. Approximately 90% of As and 36% of Pb migrated out of the RHMCS, with significant transformations observed after sintering. Adjusting the sintering parameters increased migration at long dwell times and high temperatures. These findings indicate different migration behaviors and transformations of HMs within the brick layers, emphasizing the need for cautious application and potential secondary pollution risks. A potential ecological risk index confirmed the safety of the bricks in accordance with construction material standards. Overall, this study provides crucial insights into safe and effective RHMCS utilization, contributing significantly to environmental remediation and sustainable construction practices.

Transcription factor NtMYB59 targets NtMYB12 to negatively regulate the biosynthesis of polyphenols in Nicotiana tabacum.

MYB12 is a key regulator that has been shown to promote the accumulation of various phenylpropanoid compounds in plants. However, the regulation of MYB12 gene is largely unknown. In this study, we found that overexpression of the NtMYB59 gene significantly inhibited the accumulation of chlorogenic acid (CGA), flavonols, and anthocyanins in tobacco, while knock-down and knock-out of NtMYB59 significantly increased the contents of these polyphenol compounds. Transcriptome analysis between WT and NtMYB59-OE plants revealed several differentially expressed genes (DEGs) encoding crucial enzymes in the phenylpropanoid pathway and the transcription factor NtMYB12. ChIP-seq assay further indicated that NtMYB12 might be a direct target of NtMYB59. Subsequent yeast one-hybrid, electrophoretic mobility shift assay, and Dual-Luciferase assays confirmed that NtMYB59 directly binds to the promoter of NtMYB12 to inhibit its expression. Moreover, loss-function of NtMYB59 significantly promoted the accumulation of flavonols and anthocyanins in ntmyb59, but their contents in ntmyb59/ntmyb12 double mutants were significantly lower than that of WT and ntmyb59 plants, indicating that the regulation of NtMYB59 on flavonoids biosynthesis depends on the activity of NtMYB12. Our study revealed that NtMYB59 regulates the expression of NtMYB12, and provided new potential strategies for modulating phenylpropanoids biosynthesis in tobacco.

GPR34 is a metabolic immune checkpoint for ILC1-mediated antitumor immunity.

Type 1 innate lymphoid cells (ILC1s) are a class of tissue-resident cells with antitumor activity, suggesting its possible role in solid tumor immune surveillance, but it is not clear whether manipulating ILC1s can induce potent antitumor immune responses. Here, we found that G-protein-coupled receptor 34 (GPR34), a receptor for lysophosphatidylserine (LysoPS), was highly expressed on ILC1s but not on conventional natural killer cells in the tumor microenvironment. LysoPS was enriched in the tumor microenvironment and could inhibit ILC1 activation via GPR34. Genetic deletion of LysoPS synthase Abhd16a expression in tumors or Gpr34 expression in ILC1s or antagonizing GPR34 enhanced ILC1 antitumor activity. In individuals with cancer, ABHD16A expression in tumors or GPR34 expression in ILC1s was inversely correlated with the antitumor activity of ILC1s or ILC1-like cells. Thus, our results demonstrate that manipulating ILC1s can induce potent antitumor immunity, and GPR34 is a metabolic immune checkpoint that can be targeted to develop ILC1-based immunotherapy.

Efficacy and safety of using a unilateral lower limb exoskeleton combined with conventional treatment in post-stroke rehabilitation: a randomized controlled trial.

Introduction: The incidence of hemiplegia caused by stroke is high. In particular, lower limb dysfunction affects the daily activities of patients, and lower limb robotic devices have been proposed to provide rehabilitation therapy to improve balance function in this patient population. Objective: To assess the effectiveness of the LiteStepper® unilateral lower limb exoskeleton (ULLE) combined with conventional treatment for balance function training in patients with post-stroke hemiplegia. Methods: This multicenter randomized controlled trial, conducted in the convalescent rehabilitation ward of four hospitals, involved 92 patients in their post-stroke phase. Participants were randomized into an experimental group (EG) or a conventional group (CG). The EG adopted the LiteStepper® ULLE combined with conventional treatment for 21 days. The CG underwent a standard daily rehabilitation routine for 21 days. The Berg Balance Scale (BBS), Functional Ambulation Category scale (FAC), 6-min walk test (6MWT), and Barthel Index (Barthel) were used for evaluations before and after 21 days of rehabilitative training. Results: The BBS scores in EG was significantly elevated compared to CG, exhibiting a profound statistical difference (P< 0.0001). Notably, these disparities persisted at both day 21 (P < 0.0001) and day 14 (P < 0.0047) post-intervention, underscoring the efficacy of the treatment in the EG. The EG demonstrated a markedly greater improvement in BBS scores from pre-rehabilitation to 21 days post-training, significantly outperforming the CG. Furthermore, at both day 14 and day 21, functional assessments including the FAC, 6MWT, and Barthel revealed improvements in both groups. However, the improvements in the EG were statistically significant compared to the CG at both time points: day 14 (FAC, P = 0.0377; 6MWT, P = 0.0494; Barthel, P = 0.0225) and day 21 (FAC, P = 0.0015; 6MWT, P = 0.0005; Barthel, P = 0.0004). These findings highlight the superiority of the intervention in the EG in enhancing functional outcomes. Regarding safety, the analysis revealed a solitary adverse event (AEs) related to the LiteStepper®ULLE device during the study period, affirming the combination therapy's safety profile when administered alongside conventional balance training in post-stroke hemiplegic patients. This underscores the feasibility and potential of incorporating LiteStepper®ULLE into rehabilitation protocols for this patient population. Discussion and significance: The LiteStepper® ULLE combined with conventional treatment is effective and safe for balance function training in patients with post-stroke hemiplegia.

Dual-Interphase-Stabilizing Sulfolane-Based Electrolytes for High-Voltage and High-Safety Lithium Metal Batteries.

High-voltage Li metal battery (HV-LMB) is one of the most promising energy storage technologies to achieve ultrahigh energy density. Nevertheless, electrolytes reported to date are difficult to simultaneously stabilize the Li metal anode and high-voltage cathode, especially without the assistance of expensive and corrosive high-concentration Li salts. Herein, a dual-interphase-stabilizing (DIS) and safe electrolyte that bypasses the high-concentration Li salt is reported. The electrolyte consists of high-flash-point sulfolane as solvent, molecular-orbital-engineered additives that enable stable B-F rich cathodic interphase, and unique C-F rich organic anodic interphase. The stable cycling of both Li metal anode and 4.75 V-LiCoO2 cathode in the DIS electrolyte (> 500 cycles) is demonstrated. HV-LMB pouch cells of a high energy density (435 Wh kg-1) can sustainably operate for more than 100 cycles. Moreover, the low cost and high thermal stability of the DIS electrolyte offer superior cost-effectiveness and safety for large-scale applications of HV-LMBs in the future.

Lysophospholipid headgroup size, and acyl chain length and saturation differentially affect vacuole acidification, Ca 2+ transport, and fusion.

SNARE-mediated membrane fusion is regulated by the lipid composition of the engaged bilayers. Lipid composition impacts fusion through direct protein lipid interactions or through modulating the physical properties of membranes at the site of contact, including the induction of positive curvature by lysophospholipids (LPLs). The degree of positive curvature induced is due to the length and saturation of the single acyl chain in addition to the size of the head group. Here we examined how yeast vacuole fusion and ion transport were differentially affected by changes in lysolipid properties. We found that lysophosphatidylcholine (LPC) with acyl chains containing 14-18 carbons all inhibited fusion with IC 50 values ranging from ∼40-120 µM. The monounsaturation of LPC-18:1 had no effect when compared to its saturated counterpart LPC-18:0. On the other hand, head group size played a more significant role in blocking fusion as lysophosphatidic acid (LPA)-18:1 failed to fully inhibit fusion. We also show that both Ca 2+ uptake and SNARE-dependent Ca 2+ efflux was sensitive to changes in the acyl chain length and saturation of LPCs, while LPA only affected Ca 2+ efflux. Finally, we tested these LPLs on vacuole acidification by the V-ATPase. This showed that LPC-18:0 could fully inhibit acidification whereas other LPCs had moderate effects. Again, LPA had no effect. Together these data suggest that the effects of LPLs were due to a combination of head group size and acyl chain length leading to a range in degree of positive curvature.

Advancing Proteolysis Targeting Chimera (PROTAC) Nanotechnology in Protein Homeostasis Reprograming for Disease Treatment.

Proteolysis targeting chimeras (PROTACs) represent a transformative class of therapeutic agents that leverage the intrinsic protein degradation machinery to modulate the hemostasis of key disease-associated proteins selectively. Although several PROTACs have been approved for clinical application, suboptimal therapeutic efficacy and potential adverse side effects remain challenging. Benefiting from the enhanced targeted delivery, reduced systemic toxicity, and improved bioavailability, nanomedicines can be tailored with precision to integrate with PROTACs which hold significant potential to facilitate PROTAC nanomedicines (nano-PROTACs) for clinical translation with enhanced efficacy and reduced side effects. In this review, we provide an overview of the recent progress in the convergence of nanotechnology with PROTAC design, leveraging the inherent properties of nanomaterials, such as lipids, polymers, inorganic nanoparticles, nanohydrogels, proteins, and nucleic acids, for precise PROTAC delivery. Additionally, we discuss the various categories of PROTAC targets and provide insights into their clinical translational potential, alongside the challenges that need to be addressed.

A critical role for microglia in regulating metabolic homeostasis and neural repair after spinal cord injury.

Traumatic spinal cord injury (SCI) often results in severe immune and metabolic disorders, aggravating neurological damage and inhibiting locomotor functional recovery. Microglia, as resident immune cells of the spinal cord, play crucial roles in maintaining neural homeostasis under physiological conditions. However, the precise role of microglia in regulating immune and metabolic functions in SCI is still unclear and is easily confused with that of macrophages. In this study, we pharmacologically depleted microglia to explore the role of microglia after SCI. We found that microglia are beneficial for the recovery of locomotor function. Depleting microglia disrupted glial scar formation, reducing neurogenesis and angiogenesis. Using liquid chromatography tandem mass spectrometry (LC‒MS/MS), we discovered that depleting microglia significantly inhibits lipid metabolism processes such as fatty acid degradation, unsaturated fatty acid biosynthesis, glycophospholipid metabolism, and sphingolipid metabolism, accompanied by the accumulation of multiple organic acids. Subsequent studies demonstrated that microglial depletion increased the inhibition of FASN after SCI. FASN inhibition exacerbated malonyl-CoA accumulation and significantly impeded the activity of mTORC1. Moreover, microglial depletion exacerbated the oxidative stress of neurons. In summary, our results indicate that microglia alleviate neural damage and metabolic disorders after SCI, which is beneficial for achieving optimal neuroprotection and neural repair.

Privacy Preserving Technology using Federated Learning and Blockchain in protecting against Adversarial Attacks for Retinal Imaging.

PURPOSE: Collaboration provides valuable data for robust artificial intelligence (AI) model development. Federated learning (FL) is a privacy enhancing technology that allows collaboration while respecting privacy via the development of models without raw data transfer. However state-of-the-art FL models still face challenges in non-independent and identically distributed (non-i.i.d.) health care settings and remain susceptible to privacy breaches. We propose a novel FL framework coupled with blockchain technology to address these challenges. DESIGN: Retrospective multicohort study SUBJECTS AND METHODS: 27,145 images from Singapore, China and Taiwan were used to design a novel FL aggregation method for the detection of myopic macular degeneration (MMD) from fundus photographs and macular disease from optical coherence tomography (OCT) scans in feature distribution skew and label distribution imbalance scenarios. We further performed adversarial attacks (label flipping and clean label). As proof of concept, blockchain was incorporated into FL to demonstrate secure transfer of model updates across collaborating sites. MAIN OUTCOME MEASURES: We evaluated our FL model performance in MMD and OCT classification and compared our model against state-of the-art FL and centralized models. RESULTS: Our FL model showed robust performance with areas under the receiving operating characteristic curves (AUC) of 0.868±0.009 for MMD detection and 0.970±0.012 for OCT macular disease classification. In label flipping attack, our FL model had an AUC of 0.861±0.019, similar to the centralized model (AUC 0.856± 0.015) and higher than other FL models (AUC 0.578-0.819) In clean label attack, our FL model had an AUC of 0.878±0.006 which was comparable to the centralized model (AUC 0.878±0.001) and superior to other state-of-the-art FL models with AUC of 0.529-0.838. Simulation showed that the additional time with blockchain in one global epoch was around 5 seconds. The addition of blockchain to the FL framework was feasible with a minimal impact on model development time. CONCLUSIONS: Our proposed FL algorithm overcomes the shortcoming of the traditional FL in non i.i.d. situations and remains robust to against adversarial attacks. The integration of blockchain adds further security during the transfer of model updates. Blockchain-enabled FL can be a trusted platform for collaborative health AI research.

Overlapping large facial nevus spilus and nevus of Ota: A case report diagnosed with dermoscopy and reflectance confocal microscopy.

We report a rare case of large facial hyperpigmentation in a 25-year-old female. Starting one month after birth, the patient developed a blue-brown patch on the right side of her face, interspersed with black macules and papules. As she aged, the lesion progressively enlarged and darkened, eventually covering the entire right side of her face. Dermoscopic and reflectance confocal microscopy examinations indicated nevus spilus in some areas and nevus of Ota in others, leading to a definitive diagnosis of overlapping nevus spilus and nevus of Ota. With no signs of malignant transformation on clinical or imaging examination, successful treatment was achieved using Q-switched alexandrite laser without any adverse effects. Our case underscores the critical value of dermoscopy and reflectance confocal microscopy in diagnosing rare facial pigmentary conditions, as we compare the imaging characteristics of nevus spilus, nevus of Ota, and similar conditions, alongside their clinical and histopathological correlations. Furthermore, our findings highlight the significant role of imaging examinations in monitoring malignancy and guiding treatment decisions.

Toward Strong UV-Vis-NIR Second-Harmonic Generation by Dimensionality Engineering of Zinc Thiocyanates.

The precise modulation of the spatial orientations and connection modes of primitives is vital for certain critically important optical functions for nonlinear optical (NLO) materials (specifically, second-harmonic generation (SHG) and optical bandgap); however, we are yet to achieve a sufficient level of control for the designed construction of efficient broadband NLO materials. Exploiting the changes in microscopic polarization that may result from dimensional increase, we propose herein a zero-dimensional (0D)-to-three-dimensional (3D) dimensionality-increase strategy to realize strong broadband SHG responses for the first time. The novel 3D pseudo diamond-like Zn(SCN)2 has been synthesized by removing SHG-inactive [NH4]+ counter cations and H2O molecules that are located between the adjacent discrete [Zn(SCN)4] building blocks within the 0D (NH4)2Zn(SCN)4·3H2O. The 0D-to-3D dimensionality engineering, proceeding from (NH4)2Zn(SCN)4·3H2O to Zn(SCN)2, results in significantly enhanced SHG responses and efficient broadband activity (8 × KH2PO4 @ 1064 nm, 4.18 eV bandgap for the former c.f. 2 × ß-BaB2O4 @ 380 nm, 30 × KH2PO4 @ 1064 nm, 2 × KTiOPO4 @ 2100 nm, 4.78 eV bandgap for the latter) from the UV to the NIR regions (SHG@300-1050 nm). Theoretical calculations and crystal structure analyses reveal that the coordination-bond-connected [Zn(SCN)4] building blocks within the diamond-like structure of Zn(SCN)2 are responsible for its giant broadband SHG responses.

Coupling polyethylene microplastics with other pollutants: Exploring their combined effects on plant health and technologies for mitigating toxicity.

The presence of microplastics in agricultural soils has raised concerns regarding their potential impacts on ecosystem health and plant growth. The introduction of microplastics into soil can alter its physicochemical properties, leading to adverse effects on plant development. Furthermore, the adsorption capabilities of microplastics may enhance the toxicity of soil pollutants, potentially resulting in detrimental effects on plant life. Large-sized microplastics may become adhered to root surfaces, impeding stomatal function and restricting nutrient uptake. Conversely, smaller microplastics and nano-plastics may be internalized by plants, causing cellular damage and genotoxicity. In addition, the presence of microplastics in soil can indirectly affect plant growth and development by altering the soil environment. Therefore, it is essential to investigate the potential impacts of microplastics on agricultural ecosystems and develop strategies to mitigate their effects. This review describes the adsorption power between polyethylene microplastics and pollutants (heavy metals, polycyclic aromatic hydrocarbons and antibiotics) commonly found in agricultural fields and the factors affecting the adsorption process. Additionally, the direct and indirect effects of microplastics on plants are summarized. Most of the single or combined microplastic contaminants showed negative effects on plant growth, with a few beneficial effects related to the characteristics of the microplastics and environmental factors. Currently microbial action and the application of soil conditioners or plant growth promoters can alleviate the effects of microplastics on plants to a certain extent. In light of the complex nature of soil environments, future research should concentrate on mitigate and control these interactions and the impact of compound pollution on ecosystems.