Penetrative Ionic Organic Molecular Cage Nanozyme for the Targeted Treatment of Keratomycosis.

Keratomycosis, caused by pathogenic fungi, is an intractable blinding eye disease. Corneal penetration is an essential requirement for conventional antifungal medications to address keratomycosis. Due to the distinctive anatomical and physiological structure of the cornea, the therapeutic efficacy is hampered by the inadequate penetration capacity. Despite the emergence of diverse antifungal drug delivery systems and advanced antifungal nanomaterials, it has remained challenging to achieve corneal penetration over the past decade. In the study, we fabricated a penetrative ionic organic molecular cage-based nanozyme (OMCzyme) for treating keratomycosis. The synthesis of OMCzyme involved two steps. Initially, the ionic OMC was synthesized by a [2+3] cycloimination reaction of triformylphloroglucinol and 2,3-diaminopropionic acid. Subsequently, OMCzyme was fabricated by coordination of Fe2⁺ with carboxyl anions and phenolic hydroxyls in the organic cage, and further deposition of silver nanoparticles on the surface of OMC-Fe complex. The as-prepared OMCzyme demonstrated excellent water dispersion, peroxidase-like activity, in vitro and in vivo biocompatibility, and corneal penetration. Notably, the nanozyme displayed targeted antifungal activity, effectively combating Fusarium solani with negligible cytotoxicity towards human corneal epithelial cells. The hybrid mimic was further demonstrated to be effective in treating keratomycosis in mice, indicating the potential of OMCzyme for curing fungal infectious diseases. This article is protected by copyright. All rights reserved.

Characteristics of bacterial community in eyelashes of patients with Demodex blepharitis.

BACKGROUND: Demodex blepharitis (DB) is a common disease of the ocular surface. The characteristics of the bacterial community in eyelash roots after Demodex infestation are still unknown. Knowledge of the characteristics of the bacterial community of eyelash follicles in patients with DB can provide valuable insights for guiding the diagnosis and treatment of DB. METHODS: Twenty-five patients with DB (DB group) and 21 non-DB volunteers (control group) were enrolled in the study. Eyelashes from the upper eyelid of the right eye were sampled, and 16S ribosomal DNA (rDNA) sequencing was performed to determine the V3-V4 regions of the microbial 16S rDNA gene within 1 month of infestation. The sequencing data of the two groups were analyzed and compared. The effect of the bacterium Burkholderia on the survival of Demodex mites was evaluated using Demodex obtained from 12 patients with DB other that the patients in the DB group. RESULTS: A total of 31 phyla and 862 genera were identified in the DB and control groups. The five most abundant phyla in the two groups were Proteobacteria, Firmicutes, Actinobacteria, Bacteroidetes and Cyanobacteria. The abundance of Actinomycetes was significantly higher in the DB group than in the control group. At the genus level, the five most abundant genera in the two groups were Pseudomonas, Burkholderia-Caballeronia-Paraburkholderia, Rolstonia and Acinetobacter; Clostridium sensu stricto 1 was abundant in the control group and Corynebacterium_1 was abundant in the DB group. Compared with the control group, the abundance of Burkholderia-Caballeronia-Paraburkholderia was 2.36-fold lower in the DB group. Linear discriminant analysis Effect Size (LEfSe) analysis revealed Burkholderia-Caballeronia-Paraburkholderia, SC_I_84_unclassified, Nonmyxobacteria and Succinvibrio to be the major biomarkers in the control group and Catenibacterium and Lachnospiraceae NK4A136 group to be the major biomarkers in the DB group. To explore the performance of these optimal marker models, receiver operational characteristic curve analysis was performed, and the average area under the curve value of Burkholderia-Caballeronia-Paraburkholderia was 0.7448. Burkholderia cepacia isolated from normal human eyelashes was fermented, and the Demodex mites isolated from patient eyelashes were cultured together with its fermented supernatant. The results showed that the fermentation supernatant could significantly reduce the survival time of the Demodex mites, suggesting the potential therapeutic value of this bacterium against Demodex. CONCLUSIONS: The composition of the bacterial community in the eyelashes of DB patients differed from that in eyelashes of healthy volunteers, revealing a decrease in bacterial diversity in infested eyelashes. This decrease may be related to the occurrence and development of DB. The supernatant of Burkholderia cepacia culture medium was found to inhibit the growth of Demodex in eyelash hair follicles, providing a new insight with potential applications for the clinical treatment of Demodex infestation.

Norepinephrine as the Intrinsic Contributor to Contact Lens-Induced Pseudomonas aeruginosa Keratitis.

Purpose: Contact lens wear (CLW) is one of the leading risk factors for Pseudomonas aeruginosa keratitis (PAK). However, the intrinsic factors that contribute to the high susceptibility to keratitis during CLW remain to be elucidated. CLW over an extended period can elevate corneal norepinephrine (NE) concentration. In this study, we investigated the role of NE in promoting PAK. Methods: We constructed an injury-induced PAK model and a CLW-induced PAK model to confirm the impact of NE during corneal infection. Pharmacological blockage of NE and gene knockdown mouse were used to investigate the downstream effector of NE. RNA sequencing was performed to explore the cellular alterations during NE treatment. Non-parametric Mann-Whitney U test or Kruskal-Wallis test were used to ascertain the significance (P < 0.05). Results: Supplementation of NE led to PAK even without artificial corneal injury during CLW. The effect was mediated by the ß2-adrenergic receptor (ß2-AR) in the corneal epithelium. The ß2-AR blockage by the NE antagonist ICI118,551 (ICI) or by deleting of its encoding gene Adrb2 significantly alleviated infection during CLW. Conversely, ß2-AR activation compromised the integrity of the epithelium and significantly increased the cortical plaque marker ezrin. Transcriptome analysis identified that the protective effect of ICI on the keratitis was mediated by dual-specificity phosphatases. Suramin, a Dusp5 antagonist, abrogated the protective effect of ICI. Conclusions: These data reveal a new mechanism by which NE acts as an intrinsic factor that promotes CLW-induced PAK and provide novel therapeutic targets for treating keratitis by targeting NE-ß2-AR.

Tannin coordinated nanozyme composite-based hybrid hydrogel eye drops for prophylactic treatment of multidrug-resistant Pseudomonas aeruginosa keratitis.

Pseudomonas aeruginosa infection is a severe acute suppurative ulcer that engulfs virtually the entire tissue in a short period and leads to devastating destruction. Antibiotic therapy is a common approach for the prophylaxis and treatment of P. aeruginosa infection. However, it is often associated with serious side effects, complications, and multidrug resistance. Therefore, it has been a long-standing challenge to explore safe and effective methods for controlling P. aeruginosa infection. Herein, tannin-coordinated nanozyme composite-based hybrid hydrogels (TCNH) are developed and characterized for the prophylactic treatment of P. aeruginosa and multidrug-resistant P. aeruginosa infections using mouse keratitis as the animal model. The TCNH eye drops are constructed by photoinitiated free radical polymerization of acetylated gelatin solution containing self-synthesized tannin-coordinated Co3O4/Ag nanozyme composite. The as-prepared TCNH displays good dispersibility, peroxidase-like activity and in vitro/in vivo biocompatibility. The nanozyme composite in TCNH seems to penetrate the interior of bacteria and exhibited significant broad-spectrum antibacterial activity owing to its intrinsic and nanozymic catalytic properties. Furthermore, TCNH eye drops can be successfully applied to treat P. aeruginosa and multidrug-resistant P. aeruginosa keratitis. The findings of this study reveal the potential of tannin-coordinated nanozyme composite-based hybrid hydrogel eye drops for treating infectious diseases.

Differentially Expressed Genes of Pseudomonas aeruginosa Isolates from Eyes with Keratitis and Healthy Conjunctival Sacs.

Background: Pseudomonas aeruginosa (P. aeruginosa) is the second-most common commensal bacterium in healthy conjunctival sacs. When the corneal epithelial barrier is damaged, P. aeruginosa in a healthy conjunctival sac can cause infectious keratitis, which can result in the loss of vision. This study was designed to investigate the differentially expressed genes (DEGs) of P. aeruginosa isolates from eyes with keratitis and from healthy conjunctival sacs to predict their functions and pathways through Illumina high-throughput RNA sequencing (RNA-seq). Methods: P. aeruginosa isolates from keratitis and healthy conjunctival sacs were obtained. The transcriptome profile of P. aeruginosa was characterized by a high throughput RNA-seq strategy using the Illumina HiSeq 2500 platform. The DEGs were analyzed with DESeq and validated through quantitative real-time polymerase chain reaction (PCR) and with experimental mice. GO enrichment and the KEGG pathway were also analyzed. Results: In genome-wide transcriptional analysis, 557 genes (332 upregulated and 225 downregulated) were found to be differentially expressed (fold change ≥ 2, p ≤ 0.05) in the strains from keratitis. GO enrichment analysis suggested that DEGs tended to be associated with cellular and metabolic processes. KEGG pathway analysis revealed the DEGs were typically associated with the pathways of the bacterial secretion system and pyoverdine metabolism. Eleven DEGs were validated using quantitative reverse-transcription PCR and verified with experimental mice. The results were consistent with those obtained in RNA-seq. Conclusion: The DEGs related to pilin, T2SS, T3SS, and pyoverdine metabolisms were significantly altered in the strains from keratitis. The findings may be helpful for further investigations on genes or pathways related to the pathogenesis of and therapeutic targets for P. aeruginosa keratitis.

Preparation and Evaluation of Starch Hydrogel/Contact Lens Composites as Epigallocatechin Gallate Delivery Systems for Inhibition of Bacterial Adhesion.

Microbial infections caused by wearing contact lenses has become a major health problem, so the design and development of antibacterial contact lenses has attracted widespread attention. To safely and effectively inhibit bacterial adhesion of contact lenses, we have facilely prepared epigallocatechin gallate (EGCG) loaded starch hydrogel/contact lens composites by in-situ free radical polymerization of the mixture containing 2-hydroxylethyl methacrylate, methacrylic acid and ethylene glycol dimethacrylate. The adequate transmittance of the resulting contact lenses was characterized by ultraviolet-visible spectrophotometry, and their satisfactory stability was examined using differential scanning calorimetry and thermogravimetric analysis. Whereafter, cytotoxicity and degradation experiments were performed to investigate the biocompatibility and degradability of the contact lenses. The results showed the nontoxicity and good degradability of the composites. Besides, the capacity of the contact lenses for in vitro release of EGCG was also evaluated, and the results showed that the EGCG in these contact lenses can be sustainably released for at least 14 days. Further bacterial adhesion assay suggested that the EGCG loaded starch hydrogel/contact lenses could significantly reduce the adhesion of Pseudomonas aeruginosa compared to the control. The EGCG loaded starch hydrogel/contact lens composites provide a potential intervention strategy for preventing ocular microbial infections and inhibiting bacterial keratitis.

Variable-Order Fractional Models for Wall-Bounded Turbulent Flows.

Modeling of wall-bounded turbulent flows is still an open problem in classical physics, with relatively slow progress in the last few decades beyond the log law, which only describes the intermediate region in wall-bounded turbulence, i.e., 30-50 y+ to 0.1-0.2 R+ in a pipe of radius R. Here, we propose a fundamentally new approach based on fractional calculus to model the entire mean velocity profile from the wall to the centerline of the pipe. Specifically, we represent the Reynolds stresses with a non-local fractional derivative of variable-order that decays with the distance from the wall. Surprisingly, we find that this variable fractional order has a universal form for all Reynolds numbers and for three different flow types, i.e., channel flow, Couette flow, and pipe flow. We first use existing databases from direct numerical simulations (DNSs) to lean the variable-order function and subsequently we test it against other DNS data and experimental measurements, including the Princeton superpipe experiments. Taken together, our findings reveal the continuous change in rate of turbulent diffusion from the wall as well as the strong nonlocality of turbulent interactions that intensify away from the wall. Moreover, we propose alternative formulations, including a divergence variable fractional (two-sided) model for turbulent flows. The total shear stress is represented by a two-sided symmetric variable fractional derivative. The numerical results show that this formulation can lead to smooth fractional-order profiles in the whole domain. This new model improves the one-sided model, which is considered in the half domain (wall to centerline) only. We use a finite difference method for solving the inverse problem, but we also introduce the fractional physics-informed neural network (fPINN) for solving the inverse and forward problems much more efficiently. In addition to the aforementioned fully-developed flows, we model turbulent boundary layers and discuss how the streamwise variation affects the universal curve.

Nurses' Occupational Stress and Presenteeism: The Mediating Role of Public Service Motivation and the Moderating Role of Health.

Presenteeism not only poses an economic cost to organizations but also generates reduced work efficiency and quality. The purpose of this study was to examine the connections between occupational stress, public service motivation (PSM), health, and presenteeism. A total of 981 nurses from 109 public hospitals in Jilin Province in China were enrolled in our study. Model 5 in the PROCESS micro was employed in order to verify the mediating effect of PSM and the moderating effect of nurses' health on the relationship between occupational stress and presenteeism, and simple slope analysis was used to further determine the moderating effect. Both challenge stress and hindrance stress had a positive impact on presenteeism among nurses. PSM is a mediating variable between occupational stress and presenteeism. Health moderates the path between challenge stress and presenteeism, with the association being significant for nurses with low levels of health. Future policy making should focus on preventing presenteeism by reducing excessive stress, enhancing PSM, and improving nurse health and wellness.

Hybrid natural hydrogels integrated with voriconazole-loaded microspheres for ocular antifungal applications.

Fungal keratitis is a major threat to ocular morbidity and blindness. The therapeutic efficacy of eye drops against fungal keratitis is limited by their poor bioavailability, especially in patients with corneal stromal ulcers that lead to tissue defects. Therefore, intervention measures that can control fungal infection while promoting tissue regeneration are desirable. Herein, we designed and fabricated natural antifungal hydrogels that comprise a decellularized porcine cornea (DPC), gelatin and microspheres (MCs) containing voriconazole (Vor) for the management of fungal keratitis with focal corneal stromal defects. The size and structure of the Vor-loaded MCs were characterized by scanning electron microscopy (SEM), Fourier-transform infrared (FTIR) spectroscopy and differential scanning calorimetry (DSC). The antifungal drug Vor showed that continuous release from the hybrid hydrogel system for up to 7 days and that MCs loading did not affect the mechanical properties, in vitro release profile or cytocompatibility of the hybrid hydrogel. Moreover, the Vor-loaded hydrogels exhibited excellent antifungal properties against F. solani and A. fumigatus. The efficiency of the natural antifungal hydrogel was evaluated by an ex vivo infectious rabbit corneal defect model. After 24 h, the number of fungal colony-forming units (CFU) significantly decreased in antifungal hydrogel-treated corneal tissue compared with that in non-treated corneas and corneas treated with hydrogels without Vor. The above results demonstrated that this natural hydrogel-based drug delivery system holds great promise for preventing fungal keratitis infection while promoting focal corneal stromal regeneration. Additionally, natural hybrid hydrogels might be a candidate material for use with various drugs in the effective treatment of many other ocular diseases.

Corneal epithelial injury-induced norepinephrine promotes Pseudomonas aeruginosa keratitis.

Tissue injury causes the secretion of stress hormone catecholamine and increases susceptibility to opportunistic infection. Pseudomonas aeruginosa (P. aeruginosa) is an opportunistic pathogen that is a leading cause of microbial keratitis usually associated with ocular injury or contact lens wear. However, the effect of catecholamine on P. aeruginosa induced corneal infection is unknown. Here, we test if norepinephrine (NE) would promote the progression of P. aeruginosa keratitis in mice. Adult C57BL/6 mouse corneas were scarified and then inoculated with P. aeruginosa. The content of NE was elevated in corneas after scarification and inoculation with P. aeruginosa. Then, exogenous NE was applied to the infected corneas at 24 h after inoculation; control eyes were treated with sterile saline. Topical application of NE aggravated the severity of P. aeruginosa keratitis, accompanied with the increase of clinical score, bacterial load, pathological changes, neutrophils infiltration, bacterial virulence factors and proinflammatory factors levels. In order to further verify the role of NE, N-(2-Chloroethyl)-N-ethyl-2-bromobenzylamine hydrochloride (DSP-4), a neurotoxin selected to deplete NE, was injected subconjunctivally 12 h before scarification. Pre-depletion of local NE by DSP-4 significantly alleviated the severity of corneal infection. Moreover, NE was also confirmed to increase the bacterial growth and the expression of virulence factors gene in vitro. Together, these data showed that increased corneal NE content facilitated the progression of P. aeruginosa keratitis in mice by amplifying host excessive inflammatory response and bacterial virulence. Therefore, targeting NE may provide a potential strategy for the treatment of P. aeruginosa keratitis.

Multimorbidity Analysis of 13 Systemic Diseases in Northeast China.

BACKGROUND: Multimorbidity not only affects the quality of patients' lives, but can also bring a heavy economic burden to individuals, families and society. The purpose of this study was to reveal the connections between diseases, especially the important role each disease played in the entire multimorbidity network. METHODS: A total of 1,155,734 inpatients were enrolled through multistage stratified random sampling in Jilin Province in 2017. Categorical variables were compared using the Rao-Scott-χ2 test. Weighted networks were adopted to present the complex relationships of multimorbidity. RESULTS: The distributions of the number of diseases differed significantly by gender, age and health insurance scheme (P < 0.001). Diseases of the respiratory system had the highest weight in multimorbidity in young people. Endocrine, nutritional and metabolic diseases and circulatory system diseases were often associated with other systemic diseases in middle aged and old people. CONCLUSIONS: Multimorbidity with respiratory system diseases in young people should not be overlooked. Additionally, effective prevention efforts that target endocrine, nutritional and metabolic diseases and circulatory system diseases are needed in middle aged and old people.

Identification and Characterization of Mycemycin Biosynthetic Gene Clusters in Streptomyces olivaceus FXJ8.012 and Streptomyces sp. FXJ1.235.

Mycemycins A-E are new members of the dibenzoxazepinone (DBP) family, derived from the gntR gene-disrupted deep sea strain Streptomyces olivaceus FXJ8.012Δ1741 and the soil strain Streptomyces sp. FXJ1.235. In this paper, we report the identification of the gene clusters and pathways' inference for mycemycin biosynthesis in the two strains. Bioinformatics analyses of the genome sequences of S. olivaceus FXJ8.012Δ1741 and S. sp. FXJ1.235 predicted two divergent mycemycin gene clusters, mym and mye, respectively. Heterologous expression of the key enzyme genes of mym and genetic manipulation of mye as well as a feeding study in S. sp. FXJ1.235 confirmed the gene clusters and led to the proposed biosynthetic pathways for mycemycins. To the best of our knowledge, this is the first report on DBP biosynthetic gene clusters and pathways.

Mycemycins A-E, New Dibenzoxazepinones Isolated from Two Different Streptomycetes.

Five new dibenzoxazepinone derivatives, mycemycins A-E (1-5), were isolated from the ethanol extracts of mycelia of two different streptomycetes. 1 and 2 were isolated from an acidic red soil-derived strain, Streptomyces sp. FXJ1.235, and 3-5 from a gntR gene-disrupted deep-sea strain named Streptomyces olivaceus FXJ8.012Δ1741. The structures of mycemycins were elucidated by a combination of spectroscopic analyses, including 1D- and 2D-NMR techniques.