Effect of the Sho1 gene on the pathogenicity of Candida albicans and immune function in vivo.

Objectives: Sho1, a ubiquitous membrane protein in fungi, plays a pivotal role in various physiological processes, such as osmotic stress, oxidative stress, temperature response, and virulence regulation across different fungal species. This study aimed to investigate the effect of the Sho1 gene on the pathogenicity of Candida albicans and its immune function in vivo. Materials and methods: Ninety-nine clinical strains from various infection sites were collected to investigate the expression levels of the Sho1 gene compared to its levels in the standard strain (SC5314). Sho1-knockout strains (Sho1Δ/Δ) were constructed to investigate the impact of the Sho1 gene deletion on the biofilm formation, adhesion, and flocculation abilities of C. albicans. A mouse model of systemic infection was established to evaluate the impact of Sho1 deletion on survival, organ pathology, and immune cell function, as assessed by flow cytometry. Results: The expression level of the Sho1 gene was found to be higher in clinical strains derived from sterile fluids, sputum, and secretions compared to that in the standard strains. Deletion of the Sho1 gene diminished the biofilm-formation capacity of C. albicans, leading to a sparse structure and reduced thickness, as well as diminished adhesion and flocculation abilities. Deletion of the Sho1 gene prolonged mouse survival; decreased the fungal load in the liver, kidney, and spleen; and reduced inflammatory cell infiltration into the kidney. In the spleens of mice injected with the Sho1Δ/Δ strain, a decrease was observed in the percentage of M1-type macrophages and an increase in M2-type macrophages, resulting in a decreased M1/M2 macrophage ratio. Additionally, an increase was observed in the number of Th1 cells and a decrease in the number of Th2 and Th17 cells, leading to an increased Th1/Th2 ratio. Conclusion: The Sho1 gene significantly contributes to the pathogenesis of C. albicans by influencing its biological behaviour and immune response in vivo.

Dual-antigen fusion protein vaccination induces protective immunity against Candida albicans infection in mice.

Candida albicans Is a leading cause of nosocomial bloodstream infections, particularly in immunocompromised patients. Current therapeutic strategies are insufficient, highlighting the need for effective vaccines. This study aimed to evaluate the efficacy of a dual-antigen fusion protein vaccine (AH) targeting the Als3 and Hyr1 proteins of C. albicans, using AlPO4 as an adjuvant. The AH vaccine was constructed by fusing Als317-432 and Hyr125-350 proteins, and its immunogenicity was tested in BALB/c mice and New Zealand white rabbits. Mice received three intramuscular doses of the vaccine combined with AlPO4, followed by a lethal challenge with C. albicans SC5314. Survival rates, antibody responses, cytokine production, fungal burdens, and organ pathology were assessed. The vaccine's efficacy was also validated using rabbit serum. Mice vaccinated with the AH-AlPO4 combination exhibited significantly higher antibody titers, particularly IgG and its subclasses, compared to controls (p < .001). The survival rate of vaccinated mice was 80% post-infection, significantly higher than the control group (p < .01). Vaccinated mice showed reduced fungal loads in the blood, kidneys, spleen, and liver (p < .05). Increased levels of interferon gamma and interleukin (IL)-17A were observed, indicating robust T helper (Th) 1 and Th17 cell responses. Vaccination mitigated organ damage, with kidney and liver pathology scores significantly lower than those of unvaccinated mice (p < .05). Rabbit serum with polyclonal antibodies demonstrated effective antifungal activity, confirming vaccine efficacy across species. The AH-AlPO4 vaccine effectively induced strong immune responses, reduced fungal burden, and protected against organ pathology in C. albicans infections. These findings support further development of dual-antigen vaccine strategies.

Candida, a fungus, is a major cause of bloodstream infections, especially in critical care settings. This study focused on developing a vaccine to protect against Candida infection. The vaccine targeted two key proteins, Als3p and Hyr1p, found on the surface of Candida, using a combination of these proteins. To create the vaccine, we used Als3p and Hyr1p to form a fusion protein called AH, and tested the vaccine on mice, administering it with different adjuvants (substances that enhance the immune response). The results showed that the AH vaccine, particularly when combined with the adjuvant AlPO₄, induced a strong immune response in mice. This response included the production of specific antibodies and immune cells that are crucial for defending against Candida infections. Furthermore, mice receiving the AH-AlPO₄ vaccine showed significantly better survival rates and lower levels of fungal infection compared to the control group or another experimental group. The vaccine also protected vital organs, such as the kidneys and liver, from Candida-induced damage. Additionally, we used rabbit serum to validate the efficacy of the vaccine, providing cross-species confirmation of its effectiveness. The study demonstrated the potential of the AH vaccine in eliciting robust immune responses and reducing the severity of Candida albicans infections. In summary, this research introduces a promising AH vaccine, which shows effectiveness in protecting against Candida infections. The study's innovative approach and positive results contribute to the ongoing efforts to develop vaccines against fungal infections, addressing a critical healthcare challenge. Further research is needed to explore the vaccine's long-term effectiveness and safety for potential use in clinical settings.

Interactions of graphene oxide with the microbial community of biologically active filters from a water treatment plant.

With widespread occurrence and increasing concern of emerging contaminants (CECs) in source water, biologically active filters (BAF) have been gaining acceptance in water treatment. Both BAFs and graphene oxide (GO) have been shown to be effective in treating CECs. However, studies to date have not addressed interactions between GO and microbial communities in water treatment processes such as BAFs. Therefore, in the present study, we investigated the effect of GO on the properties and microbial growth rate in a BAF system. Synthesized GO was characterized with a number of tools, including scanning electron microscopy (SEM), energy dispersive x-ray spectroscopy (EDX), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and Raman spectrometry. GO exhibited the characteristic surface functional groups (i.e., C-OH, C=O, C-O-C, and COOH), crystalline structure, and sheet-like morphology. To address the potential toxicity of GO on the microbial community, reactive oxygen species (ROS) generation was measured using nitro blue tetrazolium (NBT) assay. Results revealed that during the exponential growth phase, ROS generation was not observed in the presence of GO compared to the control batch. In fact, the adenosine triphosphate (ATP) concentrations increased in the presence of GO (25 µg/L - 1000 µg/L) compared to the control without GO. The growth rate in systems with GO exceeded the control by 20 % to 46 %. SEM images showed that GO sheets can form an effective scaffold to promote bacterial adhesion, proliferation, and biofilm formation, demonstrating its biocompatibility. Next-generation sequencing (Illumina MiSeq) was used to characterize the BAF microbial community, and high-throughput sequencing analysis confirmed the greater richness and more diverse microbial communities compared to systems without GO. This study is the first to report the effect of GO on the microbial community of BAF from a water treatment plant, which provides new insights into the potential of utilizing a bio-optimized BAF for advanced and sustainable water treatment or reuse strategies.

Determination of Volatile Halogenated Hydrocarbons in Drinking and Environmental Waters by Headspace Gas Chromatography.

Volatile halogenated hydrocarbons (VHHs) are annually produced and released into the environment, posing a threat to public health. In this study, a simple, rapid, sensitive and automated method based on headspace and gas chromatography (GC) with electron-capture detection was described for the determination of VHHs in different concentration levels in water samples. The proposed headspace GC method was initially optimized, and the optimum experimental conditions found were 10-mL water sample containing 20% w/v sodium chloride placed in a 20-mL vial and stirred at 60°C for 35 min, and then 14 VHHs were well separated on DB-35 MS capillary column with a split ratio of 12.5: 1. The limits of detection were in the low µg/L level, ranging between 0.01 and 0.6 µg/L. Finally optimized method was applied for determination 14 VHHs in drinking and environmental waters. The total mean concentrations of VHHs were 34.962, 26.183, 3.228 and 647.344 µg/L in tap water, purified water with 1-year-old filter element, seawater and effluents, respectively. However, no VHHs was detected in purified water with a new filter element. The main composition is different among different water matrix, which may be attributed to their different sources.

Caspase-4 in glioma indicates deterioration and unfavorable prognosis by affecting tumor cell proliferation and immune cell recruitment.

Gliomas are the most common malignant tumors of the central nervous system, accounting for approximately 80% of all malignant brain tumors. Accumulating evidence suggest that pyroptosis plays an essential role in the progression of cancer. Unfortunately, the effect of the pyroptosis-related factor caspase-4 (CASP4) on immunotherapy and drug therapy for tumors has not been comprehensively investigated. In this study, we systematically screened six hub genes by pooling differential pyroptosis-related genes in The Cancer Genome Atlas (TCGA) glioma data and the degree of centrality of index-related genes in the protein-protein interaction network. We performed functional and pathway enrichment analyses of the six hub genes to explore their biological functions and potential molecular mechanisms. We then investigated the importance of CASP4 using Kaplan-Meier survival analysis of glioma patients. TCGA and the Chinese Glioma Genome Atlas (CGGA) databases showed that reduced CASP4 expression leads to the potent clinical deterioration of glioma patients. Computational analysis of the effect of CASP4 on the infiltration level and recruitment of glioma immune cells revealed that CASP4 expression was closely associated with a series of tumor-suppressive immune checkpoint molecules, chemokines, and chemokine receptors. We also found that aberrant CASP4 expression correlated with chemotherapeutic drug sensitivity. Finally, analysis at the cellular and tissue levels indicated an increase in CASP4 expression in glioma, and that CASP4 inhibition significantly inhibited the proliferation of glioma cells. Thus, CASP4 is implicated as a new prognostic biomarker for gliomas with the potential to further guide immunotherapy and chemotherapy strategies for glioma patients.

Enhanced chemiluminescence resonance energy transfer using surfactant-modified AIE carbon dots.

Chemiluminescence resonance energy transfer (CRET) efficiency can be enhanced by confining CRET donors and acceptors within nanoscale spaces. However, this enhanced efficiency is often affected by uncertainties stemming from the random distribution of CRET donors and acceptors in such confined environments. In this study, a novel confined nanospace was created through the surfactant modification of carbon dots (CDs) exhibiting aggregation-induced emission (AIE) characteristics. Hydrophobic CRET donors could be effectively confined within this nanospace. The distance between the CRET donors and acceptors could be controlled by anchoring the AIE-CDs as the CRET acceptors, resulting in significantly improved CRET efficiency. Furthermore, this AIE-CDs-based CRET system was successfully applied to the detection of hydrogen peroxide (H2O2) in rainwater, showcasing its potential for practical applications.

Surface-programmed microbiome assembly in phycosphere to microplastics contamination.

Recalcitrance in microplastics accounts for ubiquitous white pollution. Of special interest are the capabilities of microorganisms to accelerate their degradation sustainably. Compared to the well-studied pure cultures in degrading natural polymers, the algal-bacterial symbiotic system is considered as a promising candidate for microplastics removal, cascading bottom-up impacts on ecosystem-scale processes. This study selected and enriched the algae-associated microbial communities hosted by the indigenous isolation Desmodesmus sp. in wastewater treatment plants with micro-polyvinyl chloride, polyethylene terephthalate, polyethylene, and polystyrene contamination. Results elaborated that multiple settled and specific affiliates were recruited by the uniform algae protagonist from the biosphere under manifold microplastic stress. Alteration of distinct chemical functionalities and deformation of polymers provide direct evidence of degradation in phycosphere under illumination. Microplastic-induced phycosphere-derived DOM created spatial gradients of aromatic protein, fulvic and humic acid-like and tryptophan components to expanded niche-width. Surface thermodynamic analysis was conducted to simulate the reciprocal and reversible interaction on algal-bacterial and phycosphere-microplastic interface, revealing the enhancement of transition to stable and irreversible aggregation for functional microbiota colonization and microplastics capture. Furthermore, pangenomic analysis disclosed the genes related to the chemotaxis and the proposed microplastics biodegradation pathway in enriched algal-bacterial microbiome, orchestrating the evidence for common synthetic polymer particles and ultimately to confirm the effectiveness and potential. The present study emphasizes the necessity for future endeavors aimed at fully leveraging the potential of algal-bacterial mutualistic systems within sustainable bioremediation strategies targeting the eradication of microplastic waste.

Comparative resistomics analysis of multidrug-resistant Chryseobacteria.

Chryseobacteria consists of important human pathogens that can cause a myriad of nosocomial infections. We isolated four multidrug-resistant Chryseobacterium bacteria from activated sludge collected at domestic wastewater treatment facilities in the New York Metropolitan area. Their genomes were sequenced with Nanopore technology and used for a comprehensive resistomics comparison with 211 Chryseobacterium genomes available in the public databases. A majority of Chryseobacteria harbor 3 or more antibiotic resistance genes (ARGs) with the potential to confer resistance to at least two types of commonly prescribed antimicrobials. The most abundant ARGs, including ß-lactam class A (blaCGA-1 and blaCIA) and class B (blaCGB-1 and blaIND) and aminoglycoside (ranA and ranB), are considered potentially intrinsic in Chryseobacteria. Notably, we reported a new resistance cluster consisting of a chloramphenicol acetyltransferase gene catB11, a tetracycline resistance gene tetX, and two mobile genetic elements (MGEs), IS91 family transposase and XerD recombinase. Both catB11 and tetX are statistically enriched in clinical isolates as compared to those with environmental origins. In addition, two other ARGs encoding aminoglycoside adenylyltransferase (aadS) and the small multidrug resistance pump (abeS), respectively, are found co-located with MGEs encoding recombinases (e.g., RecA and XerD) or transposases, suggesting their high transmissibility among Chryseobacteria and across the Bacteroidota phylum, particularly those with high pathogenicity. High resistance to different classes of ß-lactam, as well as other commonly used antimicrobials (i.e., kanamycin, gentamicin, and chloramphenicol), was confirmed and assessed using our isolates to determine their minimum inhibitory concentrations. Collectively, though the majority of ARGs in Chryseobacteria are intrinsic, the discovery of a new resistance cluster and the co-existence of several ARGs and MGEs corroborate interspecies and intergenera transfer, which may accelerate their dissemination in clinical environments and complicate efforts to combat bacterial infections.

Disease spectrum and prognostic factors in patients treated for tuberculous meningitis in Shaanxi province, China.

Background: Tuberculous meningitis (TBM) is the most severe form of tuberculosis (TB) and can be difficult to diagnose and treat. We aimed to describe the clinical presentation, diagnosis, disease spectrum, outcome, and prognostic factors of patients treated for TBM in China. Methods: A multicenter retrospective study was conducted from 2009 to 2019 enrolling all presumptive TBM patients referred to Xijing tertiary Hospital from 27 referral centers in and around Shaanxi province, China. Patients with clinical features suggestive of TBM (abnormal CSF parameters) were included in the study if they had adequate baseline information to be classified as "confirmed," "probable," or "possible" TBM according to international consensus TBM criteria and remained in follow-up. Patients with a confirmed alternative diagnosis or severe immune compromise were excluded. Clinical presentation, central nervous system imaging, cerebrospinal fluid (CSF) results, TBM score, and outcome-assessed using the modified Barthel disability index-were recorded and compared. Findings: A total of 341 presumptive TBM patients met selection criteria; 63 confirmed TBM (25 culture positive, 42 Xpert-MTB/RIF positive), 66 probable TBM, 163 possible TBM, and 49 "not TBM." Death was associated with BMRC grade III (OR = 5.172; 95%CI: 2.298-11.641), TBM score ≥ 15 (OR = 3.843; 95%CI: 1.372-10.761), age > 60 years (OR = 3.566; 95%CI: 1.022-12.442), and CSF neutrophil ratio ≥ 25% (OR = 2.298; 95%CI: 1.027-5.139). Among those with confirmed TBM, nearly one-third (17/63, 27.0%) had a TBM score < 12; these patients exhibited less classic meningitis symptoms and signs and had better outcomes compared with those with a TBM score ≥ 12. In this group, signs of disseminated/miliary TB (OR = 12.427; 95%CI: 1.138-135.758) and a higher TBM score (≥15, OR = 8.437; 95%CI: 1.328-53.585) were most strongly associated with death. Conclusion: TBM patients who are older (>60 years) have higher TBM scores or CSF neutrophil ratios, have signs of disseminated/miliary TB, and are at greatest risk of death. In general, more effort needs to be done to improve early diagnosis and treatment outcome in TBM patients.

Effect of artificial natural light on the development of myopia among primary school-age children in China: a three-year longitudinal study.

AIM: To assess the efficacy of artificial natural light in preventing incident myopia in primary school-age children. METHODS: This is a prospective, randomized control, intervention study. A total of 1840 students from 39 classes in 4 primary schools in Foshan participated in this study. The whole randomization method was adopted to include classes as a group according to 1:1 randomized control. Classrooms in the control group were illuminated by usual light, and classrooms in the intervention group were illuminated by artificial natural light. All students received uncorrected visual acuity and best-corrected visual acuity measurement, non-cycloplegic autorefraction, ocular biometric examination, slit lamp and strabismus examination. Three-year follow-up, the students underwent same procedures. Myopia was defined as spherical equivalent refraction ≤ -0.50 D and uncorrected visual acuity <20/20. RESULTS: There were 894 students in the control group and 946 students in the intervention group with a mean±SD age of 7.50±0.53y. The three-year cumulative incidence rate of myopia was 26.4% (207 incident cases among 784 eligible participants at baseline) in the control group and 21.2% (164 incident cases among 774 eligible participants at baseline) in the intervention group [difference of 5.2% (95%CI, 3.7% to 10.1%); P=0.035]. There was also a significant difference in the three-year change in spherical equivalent refraction for the control group (-0.81 D) compared with the intervention group [-0.63 D; difference of 0.18 D (95%CI, 0.08 to 0.28 D); P<0.001]. Elongation of axial length was significantly different between in the control group (0.77 mm) and the intervention group [0.72 mm; difference of 0.05 mm (95%CI, 0.01 to 0.09 mm); P=0.003]. CONCLUSION: Artificial natural light in the classroom of primary schools can result in reducing incidence rate of myopia during a period of three years.

Molecule engineering strategy of toll-like receptor 7/8 agonists designed for potentiating immune stimuli activation.

Toll-like receptor 7/8 (TLR-7/8) agonists serve as a promising class of pattern recognition receptors that effectively evoke the innate immune response, making them promising immunomodulatory agents for tumor immunotherapy. However, the uncontrollable administration of TLR-7/8 agonists frequently leads to the occurrence of severe immune-related adverse events (irAEs). Thus, it is imperative to strategically design tumor-microenvironment-associated biomarkers or exogenous stimuli responsive TLR-7/8 agonists in order to accurately evaluate and activate innate immune responses. No comprehensive elucidation has been documented thus far regarding TLR-7/8 immune agonists that are specifically engineered to enhance immune activation. In this feature article, we provide an overview of the advancements in TLR-7/8 agonists, aiming to enhance the comprehension of their mechanisms and promote the clinical progression through nanomedicine strategies. The current challenges and future directions of cancer immunotherapy are also discussed, with the hope that this work will inspire researchers to explore innovative applications for triggering immune responses through TLR-7/8 agonists.

Distinctive biotransformation and biodefluorination of 6:2 versus 5:3 fluorotelomer carboxylic acids by municipal activated sludge.

Fluorotelomer carboxylic acids (FTCAs) represent an important group of per- and polyfluoroalkyl substances (PFAS) given their high toxicity, bioaccumulation potential, and frequent detection in landfill leachates and PFAS-impacted sites. In this study, we assessed the biodegradability of 6:2 FTCA and 5:3 FTCA by activated sludges from four municipal wastewater treatment plants (WWTPs) in the New York Metropolitan area. Coupling with 6:2 FTCA removal, significant fluoride release (0.56∼1.83 F-/molecule) was evident in sludge treatments during 7 days of incubation. Less-fluorinated transformation products (TPs) were formed, including 6:2 fluorotelomer unsaturated carboxylic acid (6:2 FTUCA), perfluorohexanoic acid (PFHxA), perfluoropentanoic acid (PFPeA), and perfluorobutanoic acid (PFBA). In contrast, little fluoride (0.01∼0.09 F-/molecule) was detected in 5:3 FTCA-dosed microcosms, though 25∼68% of initially dosed 5:3 FTCA was biologically removed. This implies the dominance of "non-fluoride-releasing pathways" that may contribute to the formation of CoA adducts or other conjugates over 5:3 FTCA biotransformation. The discovery of defluorinated 5:3 FTUCA revealed the possibility of microbial attacks of the C-F bond at the γ carbon to initiate the transformation. Microbial community analysis revealed the possible involvement of 9 genera, such as Hyphomicrobium and Dechloromonas, in aerobic FTCA biotransformation. This study unraveled that biotransformation pathways of 6:2 and 5:3 FTCAs can be divergent, resulting in biodefluorination at distinctive degrees. Further research is underscored to uncover the nontarget TPs and investigate the involved biotransformation and biodefluorination mechanisms and molecular basis.

Label-free fluorescence detection of mercury ions based on thymine-mercury-thymine structure and CRISPR-Cas12a.

In this work, we developed a novel label-free fluorescent sensor for the highly sensitive detection of mercury ions (Hg2+) based on the coordination chemistry of thymine-Hg2+-thymine (T-Hg2+-T) structures and the properties of CRISPR-Cas12a systems. Most notably, two T-rich sequences (a blocker and an activator) were designed to form stable double-stranded structures in the presence of Hg2+ via the T-Hg2+-T base pairing. The formation of T-T mismatched double-stranded DNA between the blocker and the activator prevented the cleavage of G-rich sequences by Cas12a, allowing them to fold into G-quadruplex-thioflavin T complexes, resulting in significantly enhanced fluorescence. Under the optimized conditions, the developed sensor showed an excellent response for Hg2+ detection in the linear range of 0.05 to 200 nM with a detection limit of 23 pM. Moreover, this fluorescent sensor exhibited excellent selectivity and was successfully used for the detection of Hg2+ in real samples of Zhujiang river water and tangerine peel, demonstrating its potential in environmental monitoring and food safety applications.

Rapid detection of per- and polyfluoroalkyl substances (PFAS) using paper spray-based mass spectrometry.

Traditional PFAS analysis by mass spectrometry (MS) is time-consuming, as laborious sample preparation (e.g., extraction and desalting) is necessary. Herein, we report fast detection of PFAS by paper spray (PS)-based MS techniques, which employs a triangular-shaped filter paper for sample loading and ionization (≤ 3 min per sample). In this study, PS-MS was first used for direct PFAS analysis of drinking water, tap water, and wastewater. Interestingly, food package paper materials can be directly cut and examined with PS-MS for possible PFAS contamination. For samples containing salt matrices which would suppress PFAS ion signal, desalting paper spray mass spectrometry (DPS-MS), was shown to be capable of rapidly desalting, ionizing and detecting PFAS species such as per-fluorooctanoic acid (PFOA) and per-fluorosulphonic acid (PFOS). The retention of PFAS on paper substrate while salts being washed away by water is likely due to hydrophilic interaction between the PFAS polar head (e.g., carboxylic acid, sulfonic acid) with the polar filter paper cellulose surface. The DPS-MS method is highly sensitive (limits of detection:1.2-4.5 ppt) and can be applicable for directly analyzing soil extract and soil samples. These results suggest the high potential of PS-MS and the related DPS-MS technique in real-world environmental analysis of PFAS.

Insights into the roles of biochar pores toward alleviating antibiotic resistance genes accumulation in biofiltration systems.

Biofiltration systems would harbor and spread various antibiotic resistance genes (ARGs) when treating antibiotic micro-pollution, constituting a potential ecological risk. This study aimed to investigate the effects of biochar pores on ARG emergence and related microbial response mechanisms in bench-scale biofiltration systems. Results showed that biochar pores effectively reduced the absolute copies of the corresponding ARGs sul1 and sul2 by 54.1% by lowering the sorbed-SMX's bioavailability compared to non-porous anthracite. An investigation of antimicrobial resistomes revealed a considerable decrease in the abundance and diversity of ARGs and mobile gene elements. Metagenomic and metaproteomic analysis demonstrated that biochar pores induced the changeover of microbial defense strategy against SMX from blocking SMX uptake by EPS absorbing to SMX biotransformation. Microbial SOS response, antibiotic efflux pump, EPS secretion, and biofilm formation were decreased. Functions related to SMX biotransformation, such as sadABC-mediated transformation, xenobiotics degradation, and metabolism, were significantly promoted.

Angstrom-Scale Electrochemistry at Electrodes with Dimensions Commensurable and Smaller than Individual Reacting Species.

In nature and technologies, many chemical reactions occur at interfaces with dimensions approaching that of a single reacting species in nano- and angstrom-scale. Mechanisms governing reactions at this ultimately small spatial regime remain poorly explored because of challenges to controllably fabricate required devices and assess their performance in experiment. Here we report how efficiency of electrochemical reactions evolves for electrodes that range from just one atom in thickness to sizes comparable with and exceeding hydration diameters of reactant species. The electrodes are made by encapsulating graphene and its multilayers within insulating crystals so that only graphene edges remain exposed and partake in reactions. We find that limiting current densities characterizing electrochemical reactions exhibit a pronounced size effect if reactant's hydration diameter becomes commensurable with electrodes' thickness. An unexpected blockade effect is further revealed from electrodes smaller than reactants, where incoming reactants are blocked by those adsorbed temporarily at the atomically narrow interfaces. The demonstrated angstrom-scale electrochemistry offers a venue for studies of interfacial behaviors at the true molecular scale.

A syndromic diagnostic assay on a macrochannel-to-digital microfluidic platform for automatic identification of multiple respiratory pathogens.

The worldwide COVID-19 pandemic has changed people's lives and the diagnostic landscape. The nucleic acid amplification test (NAT) as the gold standard for SARS-CoV-2 detection has been applied in containing its transmission. However, there remains a lack of an affordable on-site detection system at resource-limited areas. In this study, a low cost "sample-in-answer-out" system incorporating nucleic acid extraction, purification, and amplification was developed on a single macrochannel-to-digital microfluidic chip. The macrochannel fluidic subsystem worked as a world-to-chip interface receiving 500-1000 µL raw samples, which then underwent bead-based extraction and purification processes before being delivered to DMF. Electrodes actuate an eluent dispensed to eight independent droplets for reverse transcription quantitative polymerase chain reaction (RT-qPCR). By reading with 4 florescence channels, the system can accommodate a maximum of 32 detection targets. To evaluate the proposed platform, a comprehensive assessment was conducted on the microfluidic chip as well as its functional components (i.e., extraction and amplification). The platform demonstrated a superior performance. In particular, using clinical specimens, the chip targeting SARS-CoV-2 and Flu A/B exhibited 100% agreement with off-chip diagnoses. Furthermore, the fabrication of chips is ready for scaled-up manufacturing and they are cost-effective for disposable use since they are assembled using a printed circuit board (PCB) and prefabricated blocks. Overall, the macrochannel-to-digital microfluidic platform coincides with the requirements of point-of-care testing (POCT) because of its advantages: low-cost, ease of use, comparable sensitivity and specificity, and availability for mass production.

Diffusion basis spectrum imaging detects pathological alterations in substantia nigra and white matter tracts with early-stage Parkinson's disease.

OBJECTIVES: Using diffusion basis spectrum imaging (DBSI) to examine the microstructural changes in the substantia nigra (SN) and global white matter (WM) tracts of patients with early-stage PD. METHODS: Thirty-seven age- and sex-matched patients with early-stage PD and 22 healthy controls (HCs) were enrolled in this study. All participants underwent clinical assessments and diffusion-weighted MRI scans, analyzed by diffusion tensor imaging (DTI) and DBSI to assess the pathologies of PD in SN and global WM tracts. RESULTS: The lower DTI fraction anisotropy (FA) was seen in SN of PD patients (PD: 0.316 ± 0.034 vs HCs: 0.331 ± 0.019, p = 0.015). The putative cells marker-DBSI-restricted fraction (PD: 0.132 ± 0.051 vs HCs: 0.105 ± 0.039, p = 0.031) and the edema/extracellular space marker-DBSI non-restricted-fraction (PD: 0.150 ± 0.052 vs HCs: 0.122 ± 0.052, p = 0.020) were both significantly higher and the density of axons/dendrites marker-DBSI fiber-fraction (PD: 0.718 ± 0.073 vs HCs: 0.773 ± 0.071, p = 0.003) was significantly lower in SN of PD patients. DBSI-restricted fraction in SN was negatively correlated with HAMA scores (r = - 0.501, p = 0.005), whereas DTI-FA was not correlated with any clinical scales. In WM tracts, only higher DTI axial diffusivity (AD) among DTI metrics was found in multiple WM regions in PD, while lower DBSI fiber-fraction and higher DBSI non-restricted-fraction were detected in multiple WM regions. DBSI non-restricted-fraction in both left fornix (cres)/stria terminalis (r = -0.472, p = 0.004) and right posterior thalamic radiation (r = - 0.467, p = 0.005) was negatively correlated with MMSE scores. CONCLUSION: DBSI could potentially detect and quantify the extent of inflammatory cell infiltration, fiber/dendrite loss, and edema in both SN and WM tracts in patients with early-stage PD, a finding remains to be further investigated through more extensive longitudinal DBSI analysis. CLINICAL RELEVANCE STATEMENT: Our study shows that DBSI indexes can potentially detect early-stage PD's pathological changes, with a notable ability to distinguish between inflammation and edema. This implies that DBSI has the potential to be an imaging biomarker for early PD diagnosis. KEY POINTS: • Diffusion basis spectrum imaging detected higher restricted-fraction in Parkinson's disease, potentially reflecting inflammatory cell infiltration. • Diffusion basis spectrum imaging detected higher non-restricted-fraction and lower fiber-fraction in Parkinson's disease, indicating the presence of edema and/or dopaminergic neuronal/dendritic loss. • Diffusion basis spectrum imaging metrics correlated with non-motor symptoms, suggesting its potential diagnostic role to detect early-stage PD dysfunctions.

NRF2/HO-1 axis, BIRC5, and TP53 expression in ESCC and its correlation with clinical pathological characteristics and prognosis.

BACKGROUND: Many types of cancer exhibit high nuclear factor erythroid 2-related factor 2 (NRF2), which is effective in resisting drugs and radiation. However, the role of NRF2 gene expression in predicting the prognosis of esophageal squamous cell carcinoma (ESCC) remains unclear. METHODS: The association between NRF2, heme oxygenase-1 (HO-1), baculovirus IAP repeat 5 (BIRC5), P53 gene expression levels and their relationship to immune-infiltrating cells were assessed using the Cancer Genome Atlas dataset, the Human Protein Atlas and the TISDB database. The expression of NRF2, HO-1, BIRC5, and TP53 in 118 ESCC patients was detected by immunohistochemistry, and the relationship between their expression level and clinicopathological parameters and prognosis was analyzed. RESULTS: In ESCC, NRF2 overexpression was significantly associated with Han ethnicity, lymph node metastasis, and distant metastasis. HO-1 overexpression was significantly associated with differentiation, advanced clinical staging, lymph node metastasis, nerve invasion, and distant metastasis. BIRC5 overexpression was significantly associated with Han ethnicity and lymph node metastasis. TP53 overexpression was significantly associated with Han ethnicity and T staging. The NRF2/HO-1 axis expression was positively correlated with BIRC5 and TP53. Kaplan-Meier and multivariate Cox regression analysis showed that NRF2, BIRC5, and TP53 genes co-expression was an independent prognostic risk factor. TISIDB dataset analysis showed that immune-infiltrating cells were significantly negatively correlated with NRF2 and BIRC5. CONCLUSION: NRF2, BIRC5, and TP53 axis gene expressions are predictors of poor prognosis for ESCC. The overexpression of the NRF2/HO-1/BIRC5 axis may not be related to immune-infiltrating cells.

Dysfunction of human brain network hierarchy in Parkinson's disease patients with freezing of gait.

INTRODUCTION: Hierarchy has been identified as a principle underlying the organization of human brain networks. In Parkinson's disease with freezing of gait (PD-FOG), it remains unclear whether and how the network hierarchy is disrupted. Additionally, the associations between changes in the brain network hierarchy of PD patients with FOG and clinical scales remain unclear. The aim of this study was to explore alterations in the network hierarchy of PD-FOG and their clinical relevance. METHODS: In this study, the brain network hierarchy of each group was described through a connectome gradient analysis among 31 PD-FOG, 50 PD patients without FOG (PD-NFOG), and 38 healthy controls (HC). Changes in the network hierarchy were assessed by comparing different gradient values of each network between the PD-FOG, PD-NFOG and HC groups. We further examined the relationship between dynamically changing network gradient values and clinical scales. RESULTS: For the second gradient, Salience/ventral attention network-A (SalVentAttnA) network gradient of PD-FOG group was significantly lower than that of PD-NFOG, while both PD subgroups had a Default mode network-C gradient that was significantly lower than that of the HC group. In the third gradient, somatomotor network-A gradient of PD-FOG patients was significantly lower than the PD-NFOG group. Moreover, reduced SalVentAttnA network gradient values were associated with more severe gaits, fall risk, and frozen gait in PD-FOG patients. CONCLUSIONS: The brain network hierarchy in PD-FOG is disturbed, this dysfunction is related to the severity of frozen gait. This study provides novel evidence for the neural mechanisms of FOG.