RESUMEN
The global climate is undergoing extraordinary changes, profoundly influencing a variety of ecological processes. Understanding the distribution patterns and predicting the future of plant diversity is crucial for biodiversity conservation in the context of climate change. However, current studies on predictive geographic patterns of plant diversity often fail to separate the effects of global climate change from other influencing factors. In this study, we developed a spatial simulation model of spermatophyte family diversity (SSMSFD) based on data collected from 200 nature reserves covering approximately 1,500,000 km2, where direct anthropogenic disturbances to plant diversity and the surrounding environment are absent. We predicted the spermatophyte family diversity for all provinces in China in 2020, 2040, and 2080, considering the impacts of global climate change. On average, China currently exhibits 118 plant families per 25 km2, with a decreasing trend from southeast to northwest. When considering only the effects of global climate change, excluding direct anthropogenic disturbances, our results indicate that under the Shared Socioeconomic Path Scenarios (SSPs) 245 and 585, spermatophyte family diversity is projected to slowly increase in most Chinese provinces from 2021 to 2080. Notably, the increase is more pronounced under SSPs585 compared to SSPs245. Global climate change has a positive effect on plant diversity, in contrast to the negative impact of anthropogenic disturbances that often lead to declines in plant diversity. This research highlights the contrasting outcomes of future plant diversity under the sole influence of global climate change and the significant negative effects of anthropogenic disturbances on diversity.
RESUMEN
Disinfectants and antibiotics are widely used for the prevention and control of bacterial infectious diseases. Frequent disinfection is thought to exacerbate antibiotic resistance. However, little is known about how disinfectants and antibiotics co-induce changes in the soil antibiotic resistance genes (ARGs). This study determined the ARG profiles and bacterial community dynamics between unamended soil and manure-amended soil exposed to benzalkonium chloride (C12) (BC, 10 mg kg-1) disinfectant and sulfamethazine (SMZ, 1 mg kg-1), using high-throughput quantitative PCR and 16 S rRNA gene sequencing. Manure application enriched the soil in terms of ARGs abundance and diversity, which synergistically amplified the co-selection effect of BC and SMZ on soil antibiotic resistome. Compared with the control treatment, BC and SMZ exposure had a smaller impact on the bacterial infectious diseases and antimicrobial resistance-related functions in manure-amended soil, in which bacterial communities with greater tolerance to antimicrobial substances were constructed. Manure application increased the proportion of rank I ARGs and potential human pathogenic bacteria, while BC and SMZ exposure increased the drug-resistant pathogens transmission risk. This study validated that BC and SMZ aggravated the antimicrobial resistance under manure application, providing a reference for managing the spread risk of antimicrobial resistance in agricultural activities.
Asunto(s)
Enfermedades Transmisibles , Desinfectantes , Humanos , Suelo , Antibacterianos/toxicidad , Estiércol/microbiología , Genes Bacterianos , Desinfectantes/toxicidad , Desinfectantes/análisis , Microbiología del Suelo , Bacterias/genética , SulfametazinaRESUMEN
Soil amendment with manure compost and biochar is widely adopted to improve soil fertility and promote plant growth, and their effects on soil microbial communities and resistome have been well documented. However, there is sparse information regarding their effects on vegetable endophytes, which represent a major source of human exposure to pathogens and antibiotic resistance genes (ARGs) when eaten raw. Here, we investigated the impacts of manure compost or biochar addition on the bacterial community compositions and ARGs in the soil-lettuce continuum including soil, seed, leaf, and root samples. A total of 137 ARGs and 31 mobile genetic elements (MGEs) were detected in all the samples after 60 days of cultivation. The relative abundance of ARGs and the diversity of bacteria communities presented a consistent decreasing trend from soil to root endophytes, then leaf endophytes. Manure compost addition increased the diversity and abundance of ARGs in soil, while significant changes in the ARG profiles and bacterial communities were not observed in leaf endophytes after manure compost or biochar addition, or both. Bipartite networks analysis suggested that seed microbiome was one of the major sources of plant endophytes and ARGs. Twenty potential human pathogens were isolated from lettuce, indicating potential exposure risk to pathogens via the consumption of raw lettuce. These results suggest limited impacts of manure compost and biochar addition on lettuce endophytes and highlight the contribution of seed microbiome to endophyte ARG profiles.
Asunto(s)
Endófitos , Genes Bacterianos , Humanos , Lactuca/genética , Estiércol/análisis , Microbiología del Suelo , Bacterias/genética , Suelo , Antibacterianos , Hojas de la Planta/químicaRESUMEN
Restrictions on antibiotic growth promoters have prompted livestock producers to use alternative growth promoters, and dietary copper (Cu) supplementation is currently being widely used in pig production. However, elevated doses of dietary Cu constitute a risk for co-selection of antibiotic resistance and the risk may depend on the type of Cu-based feed additives being used. We here report the first controlled experiment investigating the impact of two contrasting Cu-based feed additives on the overall swine gut microbiome and antibiotic resistome. DNA was extracted from fecal samples (n = 96) collected at four time points during 116 days from 120 pigs allotted to three dietary treatments: control, divalent copper sulfate (CuSO4; 250 µg Cu g-1 feed), and monovalent copper oxide (Cu2O; 250 µg Cu g-1 feed). Bacterial community composition, antibiotic resistance genes (ARGs), and mobile genetic elements (MGEs) were assessed, and bioavailable Cu ([Cu]bio) was determined using whole-cell bacterial bioreporters. Cu supplementation to feed increased total Cu concentrations ([Cu]total) and [Cu]bio in feces 8-10 fold and at least 670-1000 fold, respectively, but with no significant differences between the two Cu sources. The swine gut microbiome harbored highly abundant and diverse ARGs and MGEs irrespective of the treatments throughout the experiment. Microbiomes differed significantly between pig growth stages and tended to converge over time, but only minor changes in the bacterial community composition and resistome could be linked to Cu supplementation. A significant correlation between bacterial community composition (i.e., bacterial taxa present) and ARG prevalence patterns were observed by Procrustes analysis. Overall, results of the experiment did not provide evidence for Cu-induced co-selection of ARGs or MGEs even at a Cu concentration level exceeding the maximal permitted level for pig diets in the EU (25 to 150 µg Cu g-1 feed depending on pig age).
Asunto(s)
Microbioma Gastrointestinal , Porcinos , Animales , Antibacterianos/farmacología , Cobre , Farmacorresistencia Microbiana/genética , Heces , Bacterias/genética , Genes BacterianosRESUMEN
The retinal pigment epithelium (RPE) is a monolayer of polarized pigmented epithelial cells, located between the choroid and neuroretina in the retina. Multiple functions, including phagocytosis, nutrient/metabolite transportation, vitamin A metabolism, etc., are conducted by the RPE on a daily basis. RPE cells are terminally differentiated epithelial cells with little or no regenerative capacity. Loss of RPE cells results in multiple eye diseases leading to visual impairment, such as age-related macular degeneration. Therefore, the establishment of an in vitro culture model of primary RPE cells, which more closely resembles the RPE in vivo than cell lines, is critical for the characteristic and mechanistic studies of RPE cells. Considering the fact that the source of human eyeballs is limited, we create a protocol to culture primary porcine RPE cells. By using this protocol, RPE cells can be easily dissociated from adult porcine eyeballs. Subsequently, these dissociated cells attach to culture dishes/inserts, proliferate to form a confluent monolayer, and quickly re-establish key features of epithelial tissue in vivo within 2 wks. By qRT-PCR, it is demonstrated that primary porcine RPE cells express multiple signature genes at comparable levels with native RPE tissue, while the expressions of most of these genes are lost/highly reduced in human RPE-like cells, ARPE-19. Moreover, the immunofluorescence staining shows the distribution of tight junction, tissue polarity, and cytoskeleton proteins, as well as the presence of RPE65, an isomerase critical for vitamin A metabolism, in cultured primary cells. Altogether, we have developed an easy-to-follow approach to culture primary porcine RPE cells with high purity and native RPE features, which could serve as a good model to understand RPE physiology, study cell toxicities, and facilitate drug screenings.
Asunto(s)
Epitelio Pigmentado de la Retina , Vitamina A , Adulto , Animales , Porcinos , Humanos , Vitamina A/metabolismo , Retina , Células Cultivadas , Células Epiteliales , Pigmentos Retinianos/metabolismoRESUMEN
Corneal bee sting (CBS) is one of the most common ocular traumas and can lead to blindness. The ophthalmic manifestations are caused by direct mechanical effects of bee stings, toxic effects, and host immune responses to bee venom (BV); however, the underlying pathogenesis remains unclear. Clinically, topical steroids and antibiotics are routinely used to treat CBS patients but the specific drug targets are unknown; therefore, it is imperative to study the pathological characteristics, injury mechanisms, and therapeutic targets involved in CBS. In the present study, a CBS injury model was successfully established by injecting BV into the corneal stroma of healthy C57BL/6 mice. F-actin staining revealed corneal endothelial cell damage, decreased density, skeletal disorder, and thickened corneal stromal. The terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) assay showed apoptosis of both epithelial and endothelial cells. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that cytokine-cytokine interactions were the most relevant pathway for pathogenesis. Protein-protein interaction (PPI) network analysis showed that IL-1, TNF, and IL-6 were the most relevant nodes. RNA-seq after the application of Tobradex® (0.3% tobramycin and 0.1% dexamethasone) eye ointment showed that Tobradex® not only downregulated relevant inflammatory factors but also reduced corneal pain as well as promoted nerve regeneration by repairing axons. Here, a stable and reliable model of CBS injury was successfully established for the first time, and the pathogenesis of CBS and the therapeutic targets of Tobradex® are discussed. These hub genes are expected to be biomarkers and therapeutic targets for the diagnosis and treatment of CBS.
Asunto(s)
Venenos de Abeja , Lesiones de la Cornea , Mordeduras y Picaduras de Insectos , Animales , Venenos de Abeja/farmacología , Abejas/genética , Lesiones de la Cornea/diagnóstico , Lesiones de la Cornea/tratamiento farmacológico , Lesiones de la Cornea/genética , Citocinas , Modelos Animales de Enfermedad , Células Endoteliales , Ratones , Ratones Endogámicos C57BL , Combinación Dexametasona y Tobramicina , TranscriptomaRESUMEN
Ocular surface changes may develop in patients with chronic renal failure (CRF) undergoing hemodialysis. In recent years, an association of CRF with dry eye syndrome has been emphasized. However, tear proteomics of CRF patients has not been analyzed. Here, we performed systematic profiling of the tear film proteins in CRF patients through use of isobaric tags for relative and absolute quantitative (iTRAQ) MS/MS, aiming to identify associations between dry eye symptoms and expression of tear proteomic changes in patients with CRF undergoing hemodialysis. Twenty CRF patients and ten healthy subjects underwent a series of ophthalmic examinations. Tear samples from the participants were analyzed by iTRAQ approach. A total of 1139 tear proteins were screened, and 212 differentially expressed proteins were identified. The pattern changes included 77 whose expression levels were upregulated (fold increase >1.2) whereas 135 others that were downregulated (fold decrease <1/1.2). Bioinformatics analysis showed that these proteins were significantly enriched in lipid metabolism, inflammatory, and immune response pathways. Furthermore, APOA1, APOA4, APOB, APOE, S100A8, S100A9, S100A4, HSP90B and other molecules were significantly changed. Our study elucidated the characteristics of tear dynamics and protein markers in CRF patients undergoing hemodialysis. Significance: Despite the association of chronic renal failure (CRF) with dry eye disease, there are no reports describing potentially important differentially expressed tear proteins in CRF patients undergoing hemodialysis. It is still a challenge to obtain a comprehensive description of the pathogenesis of dry eye in CRF patients which hinders establishing a patient specific therapeutic scheme. Our study is the first iTRAQ proteomics analysis of the tears of patients with CRF, which reveals the changes in the protein expression profile in CRF patients afflicted with dry eye disease. The identity was verified of some relevant differentially expressed proteins, and they may be candidate diagnostic markers of dry eye disease in patients with CRF. These tear film protein constituents found in hemodialysis patients can be of important clinical significance in treating this condition. SIGNIFICANCE: Despite the association of chronic renal failure (CRF) with dry eye disease, there are no reports describing potentially important differentially expressed tear proteins in CRF patients undergoing hemodialysis. It is still a challenge to obtain a comprehensive description of the pathogenesis of dry eye in CRF patients which hinders establishing a patient specific therapeutic scheme. Our study is the first iTRAQ proteomics analysis of the tears of patients with CRF, which reveals the changes in the protein expression profile in CRF patients afflicted with dry eye disease. The identity was verified of some relevant differentially expressed proteins, and they may be candidate diagnostic markers of dry eye disease in patients with CRF. These tear film protein constituents found in hemodialysis patients can be of important clinical significance in treating this condition.
Asunto(s)
Síndromes de Ojo Seco , Fallo Renal Crónico , Síndromes de Ojo Seco/diagnóstico , Síndromes de Ojo Seco/etiología , Humanos , Fallo Renal Crónico/terapia , Proteómica , Espectrometría de Masas en Tándem , LágrimasRESUMEN
Cigarette smoke extract (CSE), a complex mixture of compounds, contributes to a range of eye diseases; however, the underlying pathophysiological responses to tobacco smoke remain ambiguous. The purpose of the present study was to evaluate the cigarette smoke-induced phenotypic and transcriptomic changes in the corneal epithelium with a view to elucidating the likely underlying mechanism. Accordingly, for the first time, we characterized the genome-wide effects of CSE on the corneal epithelium. The ocular surface of the mice in the experimental groups was exposed to CSE for 1 h per day for a period of one week, while mice in the control group were exposed to preservative-free artificial tears. Corneal fluorescein staining, in vivo confocal microscopy and scanning electron microscopy were performed to examine the corneal ultrastructure. Transcriptome sequencing and bioinformatics analysis were performed followed by RT-qPCR to validate gene expression changes. The results indicate that CSE exposure disrupted the structural integrity of the superficial epithelium, decreased the density of microvilli, and compromised the corneal epithelial barrier intactness. RNA-seq revealed 667 differentially expressed genes, and functional analysis highlighted the enhancement of several biological processes such as antioxidant activity and the response to oxidative stress. Moreover, the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis showed that glutathione metabolism and drug metabolism cytochrome P450 were the most relevant pathways contributing to the effects of CSE on the corneal epithelium. Protein-protein interaction (PPI) network analysis illustrated that GCLC, NQO1, and HMOX1 were the most relevant nodes. In conclusion, the present study indicates that CSE exposure induces changes in the phenotype and genotype of the corneal epithelium. The antioxidant response element is essential for counteracting the effects of cigarette smoke on this tissue layer. These results shed novel insights into how cigarette smoke damages this ocular surface.
Asunto(s)
Epitelio Corneal , Transcriptoma , Animales , Células Epiteliales , Ratones , Fenotipo , Humo/efectos adversos , Fumar , NicotianaRESUMEN
Homalomena occulta (Lour.) Schott (H. occulta) is a traditional Chinese medicine. However, the chloroplast genome has not been reported. Here, we assembled and analyzed the complete chloroplast (CP) genome of H. occulta. We found that the CP genome of H. occulta is 165,398 bp in length and contains a large single-copy (LSC) region of 92,861 bp, a small single-copy (SSC) region of 20,943 bp and an inverted repeat (IR) region of 25,797 bp. The genome contains 130 genes including 85 protein-coding genes, 8 rRNA and 37 tRNA. Phylogenetic analysis indicated that H. occulta is close to Philodendron lanceolatum. This study provides useful data for the development of molecular markers and identification of H. occulta.
RESUMEN
Biochar application has been a promising approach to improve soil quality but their optimal amount in improving physical and hydraulic properties remains contradictory and inconclusive. The objective of this study was to examine and propose an optimal biochar application amount in saline alkali soil considering their impact on soil physical and hydraulic properties. A three-year field experiment was conducted in the saline-alkali soils under plastic film-mulched drip irrigation in Xinjiang, China. The studied physical and hydraulic properties included bulk density, soil porosity, saturated soil water content (θs), permanent wilting point (PWP), field capacity (FC), plant available water (PAW), spatial distribution of soil water content, planar soil water storage (PSWS), and soil evaporation. The treatments included biochar application amounts of 0 (CK), 10 (B10), 50 (B50), and 100 t ha-1 (B100) in 2018. Additional two treatments with 25 t ha-1 (B25) and 30 t ha-1 (B30) were added in 2019 and 2020, respectively. A four-parameter Gaussian function was fitted to the single-peak curves of the studied hydraulic properties vs. biochar application amounts to determine the most optimal biochar application amount. The results indicated that: (1) All of the biochar treatments significantly decreased bulk density and increased soil porosity over CK; (2) B10 and B25 treatments significantly increased θs, FC, PAW, PWP, and PSWS of root zones in the film-mulched zones over CK, but reverse results were observed in the B50 and B100 treatments; (3) Daily and cumulative soil evaporation were increased in no mulch zones of all biochar treatments over CK; (4) A dose of 21.9 t ha-1 was recommended as the most optimal biochar application amount for improving physical and hydraulic properties of saline-alkali soil. This research provided useful information on biochar application amounts for improving physical and hydraulic properties in saline-alkali soil.
Asunto(s)
Álcalis , Suelo , Carbón Orgánico , ChinaRESUMEN
At present, scholars have mainly focused on the individual-level influencing factors of constructive deviance, and few studies have concerned the motivating mechanism of empowering leadership on constructive deviance. Based on the conservation of resources theory, this study explored the cross-level influence of empowering leadership on constructive deviance in the Chinese cultural context. With the data of 85 leaders and 383 paired employees which were collected in two waves with one-month time lag, the results demonstrated that empowering leadership motivated employees to actively implement constructive deviance, and that organization-based self-esteem played a mediating role in the relationship. The high traditionality of employees weakened not only the positive effect of organization-based self-esteem on constructive deviance, but also the mediating role of organization-based self-esteem. This study lays a theoretical basis and provides some practical guidance for leaders to take effective empowerment strategies to motivate employees to engage in constructive deviance.
RESUMEN
Microplastics (MPs) and antibiotic resistance genes (ARGs) are both considered emerging contaminants of increasing concern because their combined pollution poses a serious risk to the ecological environment and human health. In this study, high-throughput quantitative PCR techniques were used to investigate the diversity and abundance of ARGs in river water, to which two different microplastics (PVC and PVA) were added for aerated incubation. The results showed that ARGs in river water were diverse, and microplastics could induce more types of ARGs. Although the number and abundance of ARGs decreased in all three treatments, which were cultivated for 14 d by aeration, compared to those in non-treated samples, the total abundance of ARGs in treatments aerated with MPs were higher than those aerated without MPs, especially in the samples treated with water-soluble microplastics (PVA). Significant correlations between the abundance of ARGs and mobile genetic elements (MGEs) were observed, implying that the occurrence of MGEs may potentially affect the transmission and distribution of ARGs through horizontal gene transfer (HGT) in river water.
Asunto(s)
Antibacterianos , Plásticos , Antibacterianos/farmacología , Farmacorresistencia Microbiana/genética , Genes Bacterianos/genética , Humanos , Microplásticos , RíosRESUMEN
Plants and their extracts have been used especially in China for more than ten centuries for preventing and treating disease. However, there are only few reports describing their use in animal cell culture and tissue transplantation. In this study, onion epithelial membranes (OEM) is used as scaffolds to support cultures of a variety of cells such as fibroblasts and epithelial cells notably; they maintain the phenotypic characteristics of corneal epithelial cells. This improvement includes preservation of the proliferative potential and stemness of rabbit corneal epithelial cells (RCECs). Such an outcome suggests that this cost-effective technology warrants further evaluation to determine if OEM is a viable candidate for use as scaffolds in corneal epithelial transplantation surgery. To test this possibility, rabbit corneal epithelial cells expanded on OEM are transplanted to treat corneal epithelial defects in limbal stem cell deficient rabbits. This procedure is successful because it shortens the time required for wound healing to restore losses in corneal epithelial integrity, and forms a more compact and stratified epithelium framework than the untreated group. Ultimately, should they be proven to be effective in other relevant animal model systems, their usefulness for treating wounds in a clinical setting warrants consideration.
Asunto(s)
Limbo de la Córnea , Animales , Células Cultivadas , China , Córnea , Células Epiteliales , Cebollas , Conejos , Células MadreRESUMEN
Corneal endothelial disease is a global sight-threatening disease, and corneal transplantation using donor corneas remains the sole therapeutic option. A previous work demonstrated that N (2)-alanyl-glutamine (Ala-Gln) protected against apoptosis and cellular stress, and maintained intestinal tissue integrity. In this pursuit, the present study aimed to examine the effect of Ala-Gln in the protection of the corneal endothelium and expand its range of potential clinical applications. Mice in the control group were intracamerally irrigated with Ringers lactate injection, whereas those in the experimental group were irrigated with Ringers lactate injection containing Ala-Gln. The mean intraocular pressure increased to 44 ± 3.5 mm Hg during intracameral irrigation (normal range 10.2 ± 0.4 mmHg). In vivo confocal microscopy results showed that the addition of Ala-Gln protected the morphology, structure, and density of the corneal endothelial cells. Optical Coherence Tomography (OCT) measurements showed that corneal thickness was not significantly different between the two groups, because of the immediate corneal edema after irrigation, but the addition of Ala-Gln obviously promoted the recovery of the corneal edema. Scanning electron microscopy indicated that the corneal endothelial cells were severely ruptured and exfoliated in the Ringer's group accompanied with cellular edema, when compared with the Ala-Gln group. The intracameral irrigation using Ala-Gln protected the structure and expression of cytoskeleton and Na-K-ATPase, which exhibited a regular distribution and significantly increased expression in comparison to Ringer's group. Furthermore, Ala-Gln maintained the mitochondrial morphology and increased the activity of mitochondria. Moreover, transmission electron microscopy showed that intracameral irrigation of Ala-Gln reversed the ultrastructural changes induced by the acute ocular hypertension in mice. Our study demonstrates that the intracameral irrigation of Ala-Gln effectively maintained the corneal endothelial pump function and barrier function by protecting the mitochondrial function and preventing the rearrangement of cytoskeleton in acute ocular hypertension in mice.
RESUMEN
Multicomponent metal sulfides have been recognized as promising anode materials for lithium/sodium-ion storage given their enticing theoretical capacities. However, the simplification of synthetic processes and the construction of heterogeneous interfaces of multimetal sulfides remain great challenges. Herein, a hierarchical 1T-MoS2/carbon nanosheet decorated Co1-xS/N-doped carbon (Co1-xS/NC@MoS2/C) hollow nanofiber was designed and constructed via a one-pot hydrothermal method using a cobalt-based coordination polymer nanofiber. This nanofiber can transform in-situ into conductive N-doped carbon hollow fibers embedded with active Co1-xS nanoparticles, enabling the epitaxial growth of MoS2 nanosheets. Consequently, the Co1-xS/NC@MoS2/C composites achieve exceptional lithium/sodium-ion storage performance. Compared to MoS2/C microspheres and Co1-xS/NC hollow nanofibers alone, the Co1-xS/NC@MoS2/C hollow nanofibers deliver higher discharge capacities (1085.9 mAh g-1 for lithium-ion batteries (LIBs) and 748.5 mAh g-1 for sodium-ion batteries (SIBs) at 100 mA g-1), better capacity retention (910 mAh g-1 for LIBs and 636.5 mAh g-1 for SIBs after 150 cycles at 100 mA g-1), and increased cycling stability (407.2 mAh g-1 after 1000 cycles for SIBs at 1000 mA g-1). Furthermore, the kinetic analysis shows that the lithium/sodium-ion storage processes of the Co1-xS/NC@MoS2/C electrode are mainly controlled by pseudocapacitance behavior. The excellent electrochemical properties can thus be ascribed to the synergy of the MoS2/C nanosheets with the enlarged interlayer spacing, good conductivity of the carbon layers, and the Co1-xS nanoparticles embedded in the hollow nanofibers with extensive reaction sites.
RESUMEN
Incomplete tear film spreading and eyelid closure can cause defective renewal of the ocular surface and air exposure-induced epithelial keratopathy (EK). In this study, we characterized the role of autophagy in mediating the ocular surface changes leading to EK. Human corneal epithelial cells (HCECs) and C57BL/6 mice were employed as EK models, respectively. Transmission electron microscopy (TEM) evaluated changes in HCECs after air exposure. Each of these models was treated with either an autophagy inhibitor [chloroquine (CQ) or 3-methyladenine (3-MA)] or activator [Rapamycin (Rapa)]. Immunohistochemistry assessed autophagy-related proteins, LC3 and p62 expression levels. Western blotting confirmed the expression levels of the autophagy-related proteins [Beclin1 and mammalian target of rapamycin (mTOR)], the endoplasmic reticulum (ER) stress-related proteins (PERK, eIF2α and CHOP) and the PI3K/Akt/mTOR signalling pathway-related proteins. Real-time quantitative PCR (qRT-PCR) determined IL-1ß, IL-6 and MMP9 gene expression levels. The TUNEL assay detected apoptotic cells. TEM identified autophagic vacuoles in both EK models. Increased LC3 puncta formation and decreased p62 immunofluorescent staining and Western blotting confirmed autophagy induction. CQ treatment increased TUNEL positive staining in HCECs, while Rapa had an opposite effect. Similarly, CQ injection enhanced air exposure-induced apoptosis and inflammation in the mouse corneal epithelium, which was inhibited by Rapa treatment. Furthermore, the phosphorylation status of PERK and eIF2α and CHOP expression increased in both EK models indicating that ER stress-induced autophagy promoted cell survival. Taken together, air exposure-induced autophagy is indispensable for the maintenance of corneal epithelial physiology and cell survival.
Asunto(s)
Autofagia/fisiología , Queratitis/patología , Animales , Apoptosis/efectos de los fármacos , Apoptosis/fisiología , Autofagia/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/fisiología , Células Cultivadas , Cloroquina/farmacología , Retículo Endoplásmico/efectos de los fármacos , Retículo Endoplásmico/metabolismo , Retículo Endoplásmico/patología , Estrés del Retículo Endoplásmico/efectos de los fármacos , Estrés del Retículo Endoplásmico/fisiología , Factor 2 Eucariótico de Iniciación/metabolismo , Femenino , Humanos , Queratitis/tratamiento farmacológico , Queratitis/metabolismo , Ratones , Ratones Endogámicos C57BL , Proteínas Asociadas a Microtúbulos/metabolismo , Fosforilación/efectos de los fármacos , Fosforilación/fisiología , Transducción de Señal/efectos de los fármacos , Transducción de Señal/fisiología , Factor de Transcripción CHOP/metabolismoRESUMEN
Pterygium postoperative granuloma (PPG) is one of the common complications of pterygium surgery. In order to provide the structural features of PPG, and to further explore its pathogenetic mechanism, we analyzed clinical and pathological characteristics of 12 PPG cases. New blood vessels were observed under a slit lamp in PPG and peripheral conjunctival tissues. In vivo confocal imaging showed that there was extensive neovascularization in the stroma, accompanied by infiltration of dendritic cells and inflammatory cells. Dense fibrous structures were observed in some PPG tissues. H&E staining results confirmed neovascularization and inflammatory cells in PPG tissues. In addition, H&E staining exhibited epithelioid tissue covering some PPG tissues. The immunofluorescence results demonstrated that the PPG epithelium was negative for K19, K10 and Muc5AC. Compared with the normal conjunctiva and pterygium, the expression of collagen IV in PPG basement membrane decreased, the expression of pan-cytokeratin (PCK), claudin 4 and E-cadherin in PPG epithelium was significantly lower, while the expression of vimentin, α-SMA and Snail was significantly increased. Therefore, our results suggest that the expression of epithelial keratin markers and goblet cell specific mucin marker is downregulated in the PPG tissues, and it likely is associated with the occurrence of EMT in granulomatous tissues.
Asunto(s)
Enfermedades de la Conjuntiva/patología , Células Epiteliales/patología , Granuloma/patología , Complicaciones Posoperatorias , Pterigion/cirugía , Adulto , Biomarcadores/metabolismo , Enfermedades de la Conjuntiva/etiología , Enfermedades de la Conjuntiva/metabolismo , Neovascularización de la Córnea/patología , Sustancia Propia/irrigación sanguínea , Regulación hacia Abajo , Femenino , Fibrosis , Granuloma/etiología , Granuloma/metabolismo , Humanos , Masculino , Microscopía Confocal , Persona de Mediana EdadRESUMEN
In order to evaluate the effects of diesel exhaust particles (DEP) on the ocular surface, different concentrations (100 and 1000⯵g/ml) of DEP eye drops were administered on the mouse ocular surface for a period of 28 days. After DEP treatment, the corneal epithelial permeability to Oregon Green Dextran was studied, which increased proportionally with time. Also, the number of corneal epithelial cell layers significantly increased, which was accompanied with a high Ki67 expression. On the other hand, the number of goblet cells in the conjunctival fornix were reduced, and apoptotic cells were detected in the corneal and conjunctival epithelium by TUNEL assay in the DEP treated group, along with increased Caspase 3/8 expression. Furthermore, the number of CD4 positive cells significantly increased in the conjunctiva, while NF-κB p65 (phospho S536) expression was elevated in the cornea and also the conjunctiva. Our data revealed that the topical administration of DEP on the ocular surface in mouse disrupted the organized structure of the ocular surface and induced an inflammation of the cornea and conjunctiva.
Asunto(s)
Contaminantes Atmosféricos/toxicidad , Conjuntiva/efectos de los fármacos , Córnea/efectos de los fármacos , Emisiones de Vehículos/toxicidad , Animales , Apoptosis/efectos de los fármacos , Caspasa 3/metabolismo , Células Epiteliales/efectos de los fármacos , Etiquetado Corte-Fin in Situ , RatonesRESUMEN
Enzymatic CO2 reduction can provide value-added chemicals from greenhouse gases at ambient temperature and pressure. However, poor solubility of CO2 results in a low conversion rate. In this work, polyethylenimine (PEI) was attached onto the surface of poly(acrylic acid)-grafted (PAA-grafted) polyethylene membranes, and then, the membranes were used in an integrated process of CO2 capture and in situ hydrogenation. Modification conditions were optimized with a surface amino group density of PEI-modified membranes as the characteristic parameter, and then, SEM, FTIR, and XPS analyses were conducted. The effect of PEI-modified membranes on enzyme-catalyzed CO2 conversion to formic acid, regeneration conditions, and reusability were studied. The results show that when the grafting ratio of PAA increased, surface amino group density of PEI-modified membranes increased up to 6.00 × 10-7 mol/cm2 and then kept constant. The optimum modification time, temperature, and PEI concentration were 40 min, 40 °C, and 0.3 wt %. With the same concentration, PEI-1800 could bring more amino groups than PEI-600. SEM, FTIR, and XPS results further confirmed PEI attachment. Introduction of membrane-supported PEI with 5.86 × 10-6 mol of amino groups facilitated greatly enzymatic CO2 hydrogenation, and the initial reaction rate increased from 0.280 to 6.90 µM/min. After being regenerated in ammonia, PEI-modified membranes could be reused, and the relative reaction rate was, respectively, 88.0% and 65.7% after 5 and 10 cycles.
RESUMEN
In recent years, the application of carbonic anhydrase (CA) in CO2 removal has attracted great interest. However, obtaining high enzyme recovery activity is difficult in existing immobilization techniques. In this work, water plasma-treated poly(vinylidene fluoride) (PVDF) membranes were modified via 3-aminopropyl triethoxy silane (KH550) or γ-(2, 3-epoxypropoxy) propyl trimethoxy silane (KH560), and then CA was attached. The immobilization process was optimized, and the catalytic properties of PVDF-attached CA were characterized. The maximum activity recovery of PVDF-KH550-CA was 60%, whereas that of PVDF-KH560-CA was 33%. The Km values of PVDF-KH550-CA, PVDF-KH560-CA, and free enzyme were 9.97 ± 0.37, 12.5 ± 0.2, and 6.18 ± 0.23 mM, respectively, and their Kcat /Km values were 206 ± 2, 117 ± 5, and 488 ± 4 M-1 ·Sec-1 . PVDF-attached CA shows excellent storage stability and reusability, and their half-life values were 82 and 78 days at 4 °C. At 25 °C, they were 50 and 37 days, respectively. PVDF-KH550-CA and PVDF-KH560-CA retained approximately 85% and 72% of the initial activity after undergoing 10 cycles. In the presence of them, the generation rates of CaCO3 were 76% and 65% of the free CA system, which were 1.6 and 1.3 times that of the blank system, respectively. Its role in accelerating CO2 sequestration holds great promise for its practical application.