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1.
BMC Ophthalmol ; 24(1): 217, 2024 May 21.
Artigo em Inglês | MEDLINE | ID: mdl-38773506

RESUMO

BACKGROUND: Only seven cases of ocular Spiroplasma infection have been reported to date, all presenting as congenital cataracts with concomitant intraocular inflammation. We describe the first case of Spiroplasma infection initially presenting as a corneal infiltrate. CASE PRESENTATION: A 1-month-old girl was referred for a corneal infiltrate in the left eye. She presented in our hospital with unilateral keratouveitis. Examination showed a stromal corneal infiltrate and dense white keratic precipitates in the left eye. Herpetic keratouveitis was suspected and intravenous acyclovir therapy was initiated. Two weeks later, the inflammation in the left eye persisted and was also noticed in the right eye. Acute angle-closure glaucoma and a cataract with dilated iris vessels extending onto the anterior lens capsule developed in the left eye. The inflammation resolved after treatment with azithromycin. Iridectomy, synechiolysis and lensectomy were performed. Bacterial metagenomic sequencing (16 S rRNA) and transmission electron microscopy revealed Spiroplasma ixodetis species in lens aspirates and biopsy. Consequently, a diagnosis of bilateral Spiroplasma uveitis was made. CONCLUSIONS: In cases of congenital cataract with concomitant intraocular inflammation, Spiroplasma infection should be considered. The purpose of this case report is to raise awareness of congenital Spiroplasma infection as a cause of severe keratouveitis, cataract and angle-closure glaucoma in newborns. Performing molecular testing on lens aspirates is essential to confirm diagnosis. Systemic macrolides are suggested as the mainstay of treatment.


Assuntos
Catarata , Infecções Oculares Bacterianas , Spiroplasma , Uveíte , Humanos , Feminino , Infecções Oculares Bacterianas/diagnóstico , Infecções Oculares Bacterianas/microbiologia , Infecções Oculares Bacterianas/complicações , Catarata/congênito , Catarata/diagnóstico , Catarata/complicações , Uveíte/diagnóstico , Uveíte/microbiologia , Uveíte/complicações , Spiroplasma/isolamento & purificação , Ceratite/diagnóstico , Ceratite/microbiologia , Recém-Nascido , Antibacterianos/uso terapêutico , Lactente
2.
BMJ Case Rep ; 17(5)2024 May 06.
Artigo em Inglês | MEDLINE | ID: mdl-38719269

RESUMO

A middle-aged male patient presented with a central corneal perforation in a deep stromal infiltrate in his left eye. An emergency therapeutic penetrating keratoplasty was performed. Microbiological evaluation of the corneal scraping specimen revealed septate fungal filaments on stains. However, culture reports after 24 hours from the scraping sample and the excised half corneal button showed growth of gram-negative bacilli. This pathogen was identified as an aerobic, non-fermentative, gram-negative, bacillus by conventional microbiology and confirmed as Myroides species by the VITEK 2 Compact system (bioMérieux, Marcy l'Etoile, France). Susceptibility to chloramphenicol was noted based on which the patient was treated with topical chloramphenicol 0.5%. No recurrence of the infection was noted. This is the first reported case of corneal infection with the Myroides species of bacteria which, heretofore, have been known to cause endocarditis and urinary tract infections.


Assuntos
Infecções Oculares Fúngicas , Ceratite , Humanos , Masculino , Pessoa de Meia-Idade , Ceratite/microbiologia , Ceratite/diagnóstico , Ceratite/tratamento farmacológico , Infecções Oculares Fúngicas/microbiologia , Infecções Oculares Fúngicas/diagnóstico , Infecções Oculares Fúngicas/tratamento farmacológico , Antibacterianos/uso terapêutico , Ceratoplastia Penetrante , Cloranfenicol/uso terapêutico , Cloranfenicol/administração & dosagem , Infecções Oculares Bacterianas/microbiologia , Infecções Oculares Bacterianas/diagnóstico , Infecções Oculares Bacterianas/tratamento farmacológico , Perfuração da Córnea/microbiologia , Perfuração da Córnea/diagnóstico
3.
Mycoses ; 67(5): e13728, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38695201

RESUMO

BACKGROUND: Fungal keratitis is a severe eye infection that can result in blindness and visual impairment, particularly in developing countries. Fusarium spp. are the primary causative agents of this condition. Diagnosis of Fusarium keratitis (FK) is challenging, and delayed treatment can lead to serious complications. However, there is limited epidemiological data on FK, especially in tropical areas. OBJECTIVES: This study aimed to describe the clinical, laboratorial and epidemiological characteristics of FK in a tropical semi-arid region of Brazil. PATIENTS/METHODS: Adult patients with laboratory-confirmed FK diagnosed between October 2019 and March 2022 were evaluated. Fusarium isolates were characterized at molecular level and evaluated regarding antifungal susceptibility. RESULTS: A total of 226 clinical samples from patients suspected of keratitis were evaluated; fungal growth was detected in 50 samples (22.12%); out of which 42 were suggestive of Fusarium spp. (84%). Molecular analysis of a randomly selected set of 27 isolates identified F. solani species complex (n = 14); F. fujikuroi sensu lato (n = 6) and F. dimerum sensu lato (n = 7); a total of 10 haplotypes were identified among the strains. All but one Fusarium strains were inhibited by amphotericin B, natamycin and fluconazole. Most patients were male (71.42%; 30 out of 42), aged from 27 to 73 years old. Trauma was the most important risk factor for FK (40.47%; 17 out of 42). Patients were treated with antifungals, corticoids and antibiotics; keratoplasty and eye enucleation were also performed. CONCLUSIONS: The study provided insights into the characteristics of FK in tropical regions and emphasized the importance of enhanced surveillance and management strategies.


Assuntos
Antifúngicos , Infecções Oculares Fúngicas , Fusariose , Fusarium , Ceratite , Testes de Sensibilidade Microbiana , Humanos , Brasil/epidemiologia , Fusarium/genética , Fusarium/efeitos dos fármacos , Fusarium/isolamento & purificação , Fusarium/classificação , Masculino , Feminino , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Adulto , Ceratite/microbiologia , Ceratite/epidemiologia , Ceratite/tratamento farmacológico , Pessoa de Meia-Idade , Fusariose/microbiologia , Fusariose/epidemiologia , Fusariose/tratamento farmacológico , Infecções Oculares Fúngicas/microbiologia , Infecções Oculares Fúngicas/epidemiologia , Infecções Oculares Fúngicas/tratamento farmacológico , Idoso , Adulto Jovem , Adolescente , Clima Tropical , Idoso de 80 Anos ou mais , Anfotericina B/farmacologia , Anfotericina B/uso terapêutico
4.
Microbiology (Reading) ; 170(5)2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38739119

RESUMO

Introduction. Bacterial keratitis, particularly caused by Pseudomonas aeruginosa, is challenging to treat because of multi-drug tolerance, often associated with the formation of biofilms. Antibiotics in development are typically evaluated against planktonic bacteria in a culture medium, which may not accurately represent the complexity of infections in vivo.Hypothesis/Gap Statement. Developing a reliable, economic ex vivo keratitis model that replicates some complexity of tissue infections could facilitate a deeper understanding of antibiotic efficacy, thus aiding in the optimization of treatment strategies for bacterial keratitis.Methodology. Here we investigated the efficacy of three commonly used antibiotics (gentamicin, ciprofloxacin and meropenem) against Pseudomonas aeruginosa cytotoxic strain PA14 and invasive strain PA01 using an ex vivo porcine keratitis model.Results. Both strains of P. aeruginosa were susceptible to the MIC of the three tested antibiotics. However, significantly higher concentrations were necessary to inhibit bacterial growth in the minimum biofilm eradication concentration (MBEC) assay, with both strains tolerating concentrations greater than 512 mg l-1 of meropenem. When MIC and higher concentrations than MBEC (1024 mg l-1) of antibiotics were applied, ciprofloxacin exhibited the highest potency against both P. aeruginosa strains, followed by meropenem, while gentamicin showed the least potency. Despite this, none of the antibiotic concentrations used effectively cleared the infection, even after 18 h of continuous exposure.Conclusions. Further exploration of antibiotic concentrations and aligning dosing with clinical studies to validate the model is needed. Nonetheless, our ex vivo porcine keratitis model could be a valuable tool for assessing antibiotic efficacy.


Assuntos
Antibacterianos , Biofilmes , Ciprofloxacina , Modelos Animais de Doenças , Ceratite , Testes de Sensibilidade Microbiana , Infecções por Pseudomonas , Pseudomonas aeruginosa , Animais , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/crescimento & desenvolvimento , Antibacterianos/farmacologia , Suínos , Infecções por Pseudomonas/tratamento farmacológico , Infecções por Pseudomonas/microbiologia , Biofilmes/efeitos dos fármacos , Ceratite/microbiologia , Ceratite/tratamento farmacológico , Ciprofloxacina/farmacologia , Gentamicinas/farmacologia , Meropeném/farmacologia
5.
BMJ Case Rep ; 17(5)2024 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-38749526

RESUMO

We report the case of a female patient in her late 20s who visited the clinic with concerns about poor vision, redness, watering and a burning sensation in her left eye 2 weeks after undergoing a small incision lenticule extraction. She had no history of systemic illness or immunosuppressed status. On slit lamp examination, she was found to have corneal stromal infiltrates in the interface at multiple locations. Given the clinical diagnosis of microbial keratitis, corneal scraping of the interface infiltrate was performed and sent for microbiological examination revealing gram-positive, thin, beaded filaments that were acid-fast positive and later identified by growth in culture media as Nocardia species. This case was managed successfully with the use of topical amikacin and systemic trimethoprim-sulfamethoxazole with complete resolution of infection.


Assuntos
Antibacterianos , Infecções Oculares Bacterianas , Ceratite , Nocardiose , Humanos , Feminino , Nocardiose/diagnóstico , Nocardiose/tratamento farmacológico , Ceratite/microbiologia , Ceratite/tratamento farmacológico , Ceratite/diagnóstico , Ceratite/cirurgia , Antibacterianos/uso terapêutico , Infecções Oculares Bacterianas/diagnóstico , Infecções Oculares Bacterianas/microbiologia , Infecções Oculares Bacterianas/tratamento farmacológico , Amicacina/uso terapêutico , Amicacina/administração & dosagem , Adulto , Combinação Trimetoprima e Sulfametoxazol/uso terapêutico , Nocardia/isolamento & purificação , Infecção da Ferida Cirúrgica/microbiologia , Infecção da Ferida Cirúrgica/tratamento farmacológico , Infecção da Ferida Cirúrgica/diagnóstico
6.
Cytokine ; 179: 156626, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38678810

RESUMO

PURPOSE: To determine the antifungal, anti-inflammatory and neuroprotective effects of resveratrol (RES) in Aspergillus fumigatus (A. fumigatus) keratitis. METHODS: Cytotoxicity assay and Draize eye assay were performed to assess the toxicity of RES. The antifungal effect of RES was assessed by minimal inhibitory concentration, scanning or transmission electron microscopy, propidium iodide uptake assay, and Calcofluor white staining. Phosphorylation of p38 MAPK, mRNA and protein levels of Dectin-1 and related inflammatory factors were measured by qRT-PCR, ELISA and Western blot in vitro and in vivo. Clinical score, HE staining, plate count, and myeloperoxidase test were used to observe the progress of fungal keratitis. IF staining, qRT-PCR, and the Von Frey test were selected to assess the neuroprotective effects of RES. RESULTS: RES suppressed A. fumigatus hyphae growth and altered hyphae morphology in vitro. RES decreased the expression of Dectin-1, IL-1ß and TNF-α, as well as p38 MAPK phosphorylation expression, and also decreased clinical scores, reduced inflammatory cell infiltration and neutrophil activity, and decreased fungal load. RES also protected corneal basal nerve fibers, down-regulated mechanosensitivity thresholds, and increased the mRNA levels of CGRP and TRPV-1.. CONCLUSION: These evidences revealed that RES could exert antifungal effects on A. fumigatus and ameliorate FK through suppressing the Dectin-1/p38 MAPK pathway to down-regulate IL-1ß, IL-6, etc. expression and play protective effect on corneal nerves.


Assuntos
Anti-Inflamatórios , Aspergillus fumigatus , Ceratite , Lectinas Tipo C , Fármacos Neuroprotetores , Resveratrol , Proteínas Quinases p38 Ativadas por Mitógeno , Aspergillus fumigatus/efeitos dos fármacos , Lectinas Tipo C/metabolismo , Ceratite/tratamento farmacológico , Ceratite/metabolismo , Ceratite/microbiologia , Resveratrol/farmacologia , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Animais , Fármacos Neuroprotetores/farmacologia , Anti-Inflamatórios/farmacologia , Camundongos , Aspergilose/tratamento farmacológico , Aspergilose/metabolismo , Antifúngicos/farmacologia , Masculino , Transdução de Sinais/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Córnea/efeitos dos fármacos , Córnea/metabolismo
7.
Biomolecules ; 14(4)2024 Mar 24.
Artigo em Inglês | MEDLINE | ID: mdl-38672407

RESUMO

Purpose: To characterize microbial keratitis diversity utilizing species richness and Shannon Diversity Index. Methods: Corneal impression membrane was used to collect samples. All swabs were processed and analyzed by Biolab Laboratory (level V-SSN Excellence: ISO 9001:2015), Biolab Srl (Ascoli Piceno, Italy). DNA extraction, library preparation, and sequencing were performed in all samples. After sequencing, low-quality and polyclonal sequences were filtered out by the Ion software. At this point, we employed Kraken2 for microbial community analysis in keratitis samples. Nuclease-free water and all the reagents included in the experiment were used as a negative control. The primary outcome was the reduction in bacterial DNA (microbial load) at T1, expressed as a percentage of the baseline value (T0). Richness and Shannon alpha diversity metrics, along with Bray-Curtis beta diversity values, were calculated using the phyloseq package in R. Principal coordinate analysis was also conducted to interpret these metrics. Results: 19 samples were included in the study. The results exhibited a motley species richness, with the highest recorded value surpassing 800 species. Most of the samples displayed richness values ranging broadly from under 200 to around 600, indicating considerable variability in species count among the keratitis samples. Conclusions: A significant presence of both typical and atypical bacterial phyla in keratitis infections, underlining the complexity of the disease's microbial etiology.


Assuntos
Biodiversidade , Ceratite , Ceratite/microbiologia , Humanos , Bactérias/genética , Bactérias/classificação , Bactérias/isolamento & purificação , DNA Bacteriano/genética , Microbiota/genética
8.
Int Ophthalmol ; 44(1): 205, 2024 Apr 27.
Artigo em Inglês | MEDLINE | ID: mdl-38676784

RESUMO

PURPOSE: Microbial keratitis is a sight-threatening condition with a higher incidence in agrarian populations. In countries with a high indigent population, due to financial and other constraints, patients prefer to seek therapy locally rather than travel to advanced centres. The aim of this study is to describe the epidemiology, clinical characteristics, and outcomes of 60 consecutive patients with microbial keratitis managed at a rural centre. METHODS: Descriptive case series. All patients clinically diagnosed with infectious keratitis were included. Corneal scrapings were obtained and microbiological identification was done by Gram stain. Anti-microbial therapy was commenced based on smear findings and the patients were followed up till disease resolution. RESULTS: Sixty eyes of 60 patients were diagnosed with microbial keratitis in the study period. The mean age was 47.43 ± 18.69 years. Male:female ratio was 47:53. Risk factors included ocular trauma in the majority of patients (46/60; 76.7%). Microorganisms were identified on 75.6% of smears, with fungal filaments (65.4%) being the most common. Ulcers were central in over half (32/60; 53.3%), and > 3 mm in diameter in over three-fourths (81.6%) of patients. Forty-four patients (73.3%) achieved treatment success whereas 16/60 (26.6%) required referral to our tertiary-eye care facility for management. The median time to resolution was 14 days (IQR 10-26 days). CONCLUSION: Our series demonstrates the feasibility of microbiology-guided therapy in microbial keratitis by ophthalmologists at the secondary rural eye-care level. Two-thirds of the patients could be successfully managed at the rural centre and only severe cases needed a referral to tertiary centres.


Assuntos
Infecções Oculares Bacterianas , População Rural , Humanos , Masculino , Feminino , Pessoa de Meia-Idade , Adulto , Infecções Oculares Bacterianas/epidemiologia , Infecções Oculares Bacterianas/microbiologia , Infecções Oculares Bacterianas/diagnóstico , Infecções Oculares Bacterianas/terapia , Idoso , Índia/epidemiologia , População Rural/estatística & dados numéricos , Ceratite/epidemiologia , Ceratite/microbiologia , Ceratite/diagnóstico , Adulto Jovem , Antibacterianos/uso terapêutico , Adolescente , Úlcera da Córnea/microbiologia , Úlcera da Córnea/epidemiologia , Úlcera da Córnea/diagnóstico , Úlcera da Córnea/tratamento farmacológico , Úlcera da Córnea/terapia , Incidência , Infecções Oculares Fúngicas/epidemiologia , Infecções Oculares Fúngicas/microbiologia , Infecções Oculares Fúngicas/diagnóstico , Infecções Oculares Fúngicas/terapia , Infecções Oculares Fúngicas/tratamento farmacológico , Fatores de Risco , Bactérias/isolamento & purificação
9.
Int Immunopharmacol ; 132: 112046, 2024 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-38593508

RESUMO

PURPOSE: To investigate the potential treatment of formononetin (FMN) on Aspergillus fumigatus (A. fumigatus) keratitis with anti-inflammatory and antifungal activity. METHODS: The effects of FMN on mice with A. fumigatus keratitis were evaluated through keratitis clinical scores, hematoxylin-eosin (HE) staining, and plate counts. The expression of pro-inflammatory factors was measured using RT-PCR, ELISA, or Western blot. The distribution of macrophages and neutrophils was explored by immunofluorescence staining. The antifungal properties of FMN were assessed through minimum inhibitory concentration (MIC), propidium iodide (PI) staining, fungal spore adhesion, and biofilm formation assay. RESULTS: In A. fumigatus keratitis mice, FMN decreased the keratitis clinical scores, macrophages and neutrophils migration, and the expression of TNF-α, IL-6, and IL-1ß. In A. fumigatus-stimulated human corneal epithelial cells (HCECs), FMN reduced the expression of IL-6, TNF-α, IL-1ß, and NLRP3. FMN also decreased the expression of thymic stromal lymphopoietin (TSLP) and thymic stromal lymphopoietin receptor (TSLPR). Moreover, FMN reduced the levels of reactive oxygen species (ROS) induced by A. fumigatus in HCECs. Furthermore, FMN inhibited A. fumigatus growth, prevented spore adhesion and disrupted fungal biofilm formation in vitro. In vivo, FMN treatment reduced the fungal load in mice cornea at 3 days post infection (p.i.). CONCLUSION: FMN demonstrated anti-inflammatory and antifungal properties, and exhibited a protective effect on mouse A. fumigatus keratitis.


Assuntos
Anti-Inflamatórios , Aspergilose , Aspergillus fumigatus , Isoflavonas , Ceratite , Animais , Aspergillus fumigatus/efeitos dos fármacos , Ceratite/tratamento farmacológico , Ceratite/microbiologia , Ceratite/imunologia , Aspergilose/tratamento farmacológico , Aspergilose/imunologia , Isoflavonas/farmacologia , Isoflavonas/uso terapêutico , Humanos , Camundongos , Anti-Inflamatórios/uso terapêutico , Anti-Inflamatórios/farmacologia , Citocinas/metabolismo , Antifúngicos/uso terapêutico , Antifúngicos/farmacologia , Neutrófilos/imunologia , Neutrófilos/efeitos dos fármacos , Modelos Animais de Doenças , Espécies Reativas de Oxigênio/metabolismo , Feminino , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Biofilmes/efeitos dos fármacos , Camundongos Endogâmicos C57BL , Córnea/patologia , Córnea/efeitos dos fármacos , Córnea/microbiologia
10.
BMJ Open Ophthalmol ; 9(1)2024 Apr 22.
Artigo em Inglês | MEDLINE | ID: mdl-38653537

RESUMO

OBJECTIVE: Microbial keratitis (MK) is a significant cause of blindness in sub-Saharan Africa. We investigated the feasibility of using a novel corneal impression membrane (CIM) for obtaining and processing samples by culture, PCR and whole-genome sequencing (WGS) in patients presenting with suspected MK in Malawi. METHODS AND ANALYSIS: Samples were collected from patients presenting with suspected MK using a 12 mm diameter polytetrafluoroethylene CIM disc. Samples were processed using culture and PCR for Acanthamoeba, herpes simplex virus type 1 (HSV-1) and the bacterial 16S rRNA gene. Minimum inhibitory concentrations of isolates to eight antimicrobials were measured using susceptibility strips. WGS was used to characterise Staphylococcus aureus isolates. RESULTS: 71 eyes of 71 patients were included. The overall CIM isolation rate was 81.7% (58 positive samples from 71 participants). 69 (81.2%) of isolates were Gram-positive cocci. Coagulase-negative Staphylococcus 31.8% and Streptococcus species 14.1% were the most isolated bacteria. Seven (9.9%) participants were positive for HSV-1. Fungi and Acanthamoeba were not detected. Moxifloxacin and chloramphenicol offered the best coverage for both Gram-positive and Gram-negative isolates when susceptibility was determined using known antimicrobial first quartile concentrations and European Committee on Antimicrobial Susceptibility Testing breakpoints, respectively. WGS identified known virulence genes associated with S. aureus keratitis. CONCLUSIONS: In a resource-poor setting, a CIM can be used to safely sample the cornea in patients presenting with suspected MK, enabling identification of causative microorganisms by culture and PCR. Although the microbiological spectrum found was limited to the dry season, these preliminary results could be used to guide empirical treatment.


Assuntos
Infecções Oculares Bacterianas , Humanos , Projetos Piloto , Malaui/epidemiologia , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Infecções Oculares Bacterianas/microbiologia , Infecções Oculares Bacterianas/epidemiologia , Infecções Oculares Bacterianas/tratamento farmacológico , Adulto Jovem , Bactérias/isolamento & purificação , Bactérias/efeitos dos fármacos , Bactérias/genética , Testes de Sensibilidade Microbiana , Córnea/microbiologia , Ceratite/microbiologia , Ceratite/tratamento farmacológico , Ceratite/epidemiologia , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Idoso , Reação em Cadeia da Polimerase , Adolescente , Acanthamoeba/isolamento & purificação , Acanthamoeba/genética , Acanthamoeba/efeitos dos fármacos , RNA Ribossômico 16S/genética
11.
Int J Pharm ; 656: 124118, 2024 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-38615806

RESUMO

Fungal infections of cornea are important causes of blindness especially in developing nations with tropical climate. However, the challenges associated with current treatments are responsible for poor outcome. Natamycin is the only FDA-approved antifungal drug to treat fungal keratitis, but unfortunately due to its poor water solubility, it is available as suspension. The marketed suspension (5% Natamycin) has rapid precorneal clearance, poor corneal permeability, a higher frequency of administration, and corneal irritation due to undissolved suspended drug particles. In our study, we developed clear and stable natamycin-loaded nanomicelles (1% Natcel) to overcome the above challenges. We demonstrated that 1% Natcel could permeate the cornea better than 5% suspension. The developed 1% Natcel was able to provide sustained release for up to 24 h. Further, it was found to be biocompatible and also improved the mean residence time (MRT) than 5% suspension in tears. Therefore, the developed 1% Natcel could be a potential alternative treatment for fungal keratitis.


Assuntos
Antifúngicos , Córnea , Liberação Controlada de Fármacos , Infecções Oculares Fúngicas , Ceratite , Micelas , Nanopartículas , Natamicina , Natamicina/administração & dosagem , Antifúngicos/administração & dosagem , Antifúngicos/química , Antifúngicos/farmacologia , Ceratite/tratamento farmacológico , Ceratite/microbiologia , Animais , Córnea/microbiologia , Córnea/metabolismo , Córnea/efeitos dos fármacos , Infecções Oculares Fúngicas/tratamento farmacológico , Infecções Oculares Fúngicas/microbiologia , Coelhos , Solubilidade , Preparações de Ação Retardada , Lágrimas/metabolismo
12.
Biomaterials ; 308: 122565, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38603823

RESUMO

As bacterial keratitis progresses rapidly, prompt intervention is necessary. Current diagnostic processes are time-consuming and invasive, leading to improper antibiotics for treatment. Therefore, innovative strategies for diagnosing and treating bacterial keratitis are urgently needed. In this study, Cu2-xSe@BSA@NTRP nanoparticles were developed by loading nitroreductase-responsive probes (NTRPs) onto Cu2-xSe@BSA. These nanoparticles exhibited integrated fluorescence imaging and antibacterial capabilities. In vitro and in vivo experiments showed that the nanoparticles produced responsive fluorescence signals in bacteria within 30 min due to an interaction between the released NTRP and bacterial endogenous nitroreductase (NTR). When combined with low-temperature photothermal therapy (PTT), the nanoparticles effectively eliminated E. coli and S. aureus, achieved antibacterial efficacy above 95% and facilitated the re-epithelialization process at the corneal wound site in vivo. Overall, the Cu2-xSe@BSA@NTRP nanoparticles demonstrated potential for rapid, noninvasive in situ diagnosis, treatment, and visualization assessment of therapy effectiveness in bacterial keratitis.


Assuntos
Antibacterianos , Escherichia coli , Ceratite , Nanopartículas , Nitrorredutases , Animais , Nitrorredutases/metabolismo , Antibacterianos/farmacologia , Antibacterianos/química , Antibacterianos/uso terapêutico , Nanopartículas/química , Ceratite/tratamento farmacológico , Ceratite/microbiologia , Escherichia coli/efeitos dos fármacos , Imagem Óptica/métodos , Staphylococcus aureus/efeitos dos fármacos , Camundongos , Terapia Fototérmica/métodos , Humanos , Cobre/química
15.
Infect Immun ; 92(4): e0048323, 2024 Apr 09.
Artigo em Inglês | MEDLINE | ID: mdl-38501672

RESUMO

Aspergillus fumigatus (A. fumigatus) is one of the common pathogens of fungal keratitis. Fungal growth and invasion cause excessive inflammation and corneal damage, leading to severe vision loss. Neutrophils are the primary infiltrating cells critical for fungal clearance. Cathelicidin [LL-37 in humans and cathelicidin-related antimicrobial peptide (CRAMP) in mice], a natural antimicrobial peptide, can directly inhibit the growth of many pathogens and regulate immune responses. However, the role of cathelicidin and its effect on neutrophils in A. fumigatus keratitis remain unclear. By establishing A. fumigatus keratitis mouse models, we found that cathelicidin was increased in A. fumigatus keratitis. It could reduce fungal loads, lower clinical scores, and improve corneal transparency. Restriction of CRAMP on fungal proliferation was largely counteracted in CD18-/- mice, in which neutrophils cannot migrate into infected sites. When WT neutrophils were transferred into CD18-/- mice, corneal fungal loads were distinctly reduced, indicating that neutrophils are vital for CRAMP-mediated resistance. Furthermore, cathelicidin promoted neutrophils to phagocytose and degrade conidia both in vitro and in vivo. CXC chemokine receptor 2 (CXCR2) was reported to be a functional receptor of LL-37 on neutrophils. CXCR2 antagonist SB225002 or phospholipase C (PLC) inhibitor U73122 weakened LL-37-induced phagocytosis. Meanwhile, LL-37 induced PLC γ phosphorylation, which was attenuated by SB225002. SB225002 or the autophagy inhibitors Bafilomycin-A1 and 3-Methyladenine weakened LL-37-induced degradation of conidia. Transmission electron microscopy (TEM) observed that LL-37 increased autophagosomes in Aspergillus-infected neutrophils. Consistently, LL-37 elevated autophagy-associated protein expressions (Beclin-1 and LC3-II), but this effect was weakened by SB225002. Collectively, cathelicidin reduces fungal loads and improves the prognosis of A. fumigatus keratitis. Both in vitro and in vivo, cathelicidin promotes neutrophils to phagocytose and degrade conidia. LL-37/CXCR2 activates PLC γ to amplify neutrophils' phagocytosis and induces autophagy to eliminate intracellular conidia.


Assuntos
Aspergillus fumigatus , Ceratite , Compostos de Fenilureia , Humanos , Animais , Camundongos , Neutrófilos , Antifúngicos/metabolismo , Catelicidinas , Fosfolipase C gama/metabolismo , Ceratite/microbiologia , Prognóstico , Camundongos Endogâmicos C57BL
16.
Exp Eye Res ; 242: 109863, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38494102

RESUMO

PURPOSE: Pseudomonas aeruginosa-induced keratitis is one of the most severe and challenging forms of corneal infection, owing to its associated intense inflammatory reactions leading to corneal necrosis and dense corneal scar with loss of vision. Since mesenchymal stem cells (MSCs) are reported to possess antimicrobial and immunomodulatory properties, they can be tested as an adjuvant treatment along with the antibiotics which are the current standard of care. This study aims to investigate the anti-bacterial and immunomodulatory roles of human bone marrow MSC-derived conditioned medium (MSC-CM) in P. aeruginosa-infected human corneal epithelial cells (HCECs) in vitro. METHODS: The effect of MSC-CM on the growth of clinical isolates of P. aeruginosa was evaluated by colony-forming unit assay. The expression of inflammatory cytokines (IL-6 and TNF-α) and an antimicrobial peptide (Lipocalin 2) in lipopolysaccharide-treated MSCs and HCECs was analyzed through ELISA. Corneal epithelial repair following infection with P. aeruginosa was studied through scratch assay. RESULTS: Compared to control (P. aeruginosa (5*105) incubated in DMEM (1 ml) at 37 °C for 16 h), MSC-CM significantly: i) inhibits the growth of P. aeruginosa (159*109 vs. 104*109 CFU/ml), ii) accelerates corneal epithelial repair following infection with P. aeruginosa (9% vs. 24% closure of the wounded area after 12 h of infection), and iii) downregulates the lipopolysaccharide-induced expression of IL-6, TNF-α and Lipocalin 2 in HCECs. A combination of MSC-CM with an antibiotic, Ciprofloxacin moderately regulated the expression of IL-6, TNF-α, and Lipocalin 2. CONCLUSION: MSC-CM holds promise as an adjunctive therapeutic approach for P. aeruginosa-induced corneal epithelial damage.


Assuntos
Ensaio de Imunoadsorção Enzimática , Infecções Oculares Bacterianas , Células-Tronco Mesenquimais , Infecções por Pseudomonas , Pseudomonas aeruginosa , Humanos , Infecções Oculares Bacterianas/microbiologia , Infecções Oculares Bacterianas/metabolismo , Infecções Oculares Bacterianas/patologia , Infecções por Pseudomonas/microbiologia , Infecções por Pseudomonas/terapia , Infecções por Pseudomonas/tratamento farmacológico , Células-Tronco Mesenquimais/metabolismo , Epitélio Corneano/microbiologia , Epitélio Corneano/patologia , Epitélio Corneano/metabolismo , Células Cultivadas , Ceratite/microbiologia , Ceratite/metabolismo , Ceratite/patologia , Transplante de Células-Tronco Mesenquimais/métodos , Meios de Cultivo Condicionados/farmacologia , Estudo de Prova de Conceito , Interleucina-6/metabolismo , Úlcera da Córnea/microbiologia , Úlcera da Córnea/metabolismo , Úlcera da Córnea/patologia , Úlcera da Córnea/tratamento farmacológico , Lipocalina-2/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
17.
Mycopathologia ; 189(2): 28, 2024 Mar 14.
Artigo em Inglês | MEDLINE | ID: mdl-38483684

RESUMO

BACKGROUND: Fungal keratitis (FK) is a kind of infectious keratopathy with a high rate of blindness worldwide. Deoxynivalenol (DON) has been proven to have multiple toxic effects on humans and animals. OBJECTIVES: The aim of this study was to explore a possible pathogenic role of DON in FK. METHODS: We first made an animal model of FK in New Zealand white rabbits, and then attempted to detect DON in a culture medium in which Fusarium solani had been grown and also in the corneal tissue of the animal model of Fusarium solani keratitis. Next, a model of DON damage in human corneal epithelial cells (HCECs) was constructed to evaluate effects of DON on the activity, migration ability, cell cycle, and apoptosis in the HCECs. Then, putative the toxic damaging effects of DON on rabbit corneal epithelial cells and the impact of the repair cycle were studied. The expression levels of inflammatory factors in the corneas of the animal model and in the model of DON-damaged HCECs were measured. RESULTS: The Fusarium solani strain used in this study appeared to have the potential to produce DON, since DON was detected in the corneal tissue of rabbits which had been inoculated with this Fusarium solani strain. DON was found to alter the morphology of HCECs, to reduce the activity and to inhibit the proliferation and migration of HCECs. DON also induced the apoptosis and S-phase arrest of HCECs. In addition, DON was found to damage rabbit corneal epithelial cells, to prolong the corneal epithelial regeneration cycle, and to be associated with the upregulated expression of inflammatory factors in HCECs and rabbit corneas. CONCLUSIONS: DON appears to have a toxic damaging effect on HCECs in FK, and to induce the expression of inflammatory factors, leading to the exacerbation of keratitis and the formation of new blood vessels. Future studies will explore the possibility of developing a test to detect DON in ophthalmic settings to aid the rapid diagnosis of FK, and to develop DON neutralizers and adsorbents which have the potential to improve keratocyte status, inhibit apoptosis, and alleviate inflammation, therein providing new thinking for therapy of clinical FK.


Assuntos
Úlcera da Córnea , Infecções Oculares Fúngicas , Fusarium , Ceratite , Tricotecenos , Humanos , Coelhos , Animais , Ceratite/microbiologia , Células Epiteliais
18.
Nano Lett ; 24(13): 4044-4053, 2024 Apr 03.
Artigo em Inglês | MEDLINE | ID: mdl-38517749

RESUMO

Fungal keratitis (FK) is an infectious eye disease that poses a significant risk of blindness. However, the effectiveness of conventional antifungal drugs is limited due to the intrinsic ocular barrier that impedes drug absorption. There is an urgent need to develop new therapeutic strategies to effectively combat FK. Herein, we synthesized an ultrasmall positively charged carbon dot using a simple stage-melting method. The carbon dot can penetrate the corneal barrier by opening the tight junctions, allowing them to reach the lesion site and effectively kill the fungi. The results both in vitro and in vivo demonstrated that it exhibited good biocompatibility and antifungal activity, significantly improving the therapeutic effect in a mouse model of FK. Therefore, this biophilic ultrasmall size and positive carbon dot, characterized by its ability to penetrate the corneal barrier and its antifungal properties, may offer valuable insights into the design of effective ocular nanomedicines.


Assuntos
Úlcera da Córnea , Infecções Oculares Fúngicas , Ceratite , Animais , Camundongos , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Ceratite/tratamento farmacológico , Ceratite/microbiologia , Úlcera da Córnea/tratamento farmacológico , Úlcera da Córnea/microbiologia , Infecções Oculares Fúngicas/tratamento farmacológico , Infecções Oculares Fúngicas/microbiologia , Córnea/microbiologia
19.
J Control Release ; 368: 483-497, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38458571

RESUMO

Fungal keratitis is a refractory eye disease that is prone to causing blindness. Fungal virulence and inflammatory responses are two major factors that accelerate the course of fungal keratitis. However, the current antifungal drugs used for treatment usually possess transient residence time on the ocular surface and low bioavailability deficiencies, which limit their therapeutic efficacy. In this work, natamycin (NATA)-loaded mesoporous zinc oxide (Meso-ZnO) was synthesized for treating Aspergillus fumigatus keratitis with excellent drug-loading and sustained drug release capacities. In addition to being a carrier for drug delivery, Meso-ZnO could restrict fungal growth in a concentration-dependent manner, and the transcriptome analysis of fungal hyphae indicated that it inhibited the mycotoxin biosynthesis, oxidoreductase activity and fungal cell wall formation. Meso-ZnO also promoted cell migration and exhibited anti-inflammatory role during fungal infection by promoting the activation of autophagy. In mouse models of fungal keratitis, Meso-ZnO/NATA greatly reduced corneal fungal survival, alleviated tissue inflammatory damage, and reduced neutrophils accumulation and cytokines expression. This study suggests that Meso-ZnO/NATA can be a novel and effective treatment strategy for fungal keratitis.


Assuntos
Aspergilose , Infecções Oculares Fúngicas , Ceratite , Óxido de Zinco , Animais , Camundongos , Antifúngicos/uso terapêutico , Antifúngicos/farmacologia , Óxido de Zinco/uso terapêutico , Aspergilose/tratamento farmacológico , Aspergilose/microbiologia , Ceratite/tratamento farmacológico , Ceratite/metabolismo , Ceratite/microbiologia , Natamicina/uso terapêutico , Infecções Oculares Fúngicas/tratamento farmacológico , Infecções Oculares Fúngicas/metabolismo , Infecções Oculares Fúngicas/microbiologia , Sistemas de Liberação de Medicamentos , Camundongos Endogâmicos C57BL
20.
Indian J Med Microbiol ; 48: 100558, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38458337

RESUMO

Lasiodiplodia theobromae is a dematiaceous fungus which rarely causes keratitis and is mostly resistant to the commonly used antifungal drugs. Here, we report three cases of keratitis caused by L.theobromae from Assam. All the cases were successfully treated with 1% voriconazole and surgical debridement. To the best of our knowledge and literature search, this is the first case series of keratitis caused by L.theobromae reported from eastern India.


Assuntos
Antifúngicos , Ascomicetos , Ceratite , Voriconazol , Humanos , Voriconazol/uso terapêutico , Antifúngicos/uso terapêutico , Ceratite/tratamento farmacológico , Ceratite/microbiologia , Índia , Masculino , Ascomicetos/isolamento & purificação , Ascomicetos/efeitos dos fármacos , Feminino , Adulto , Pessoa de Meia-Idade , Resultado do Tratamento , Micoses/tratamento farmacológico , Micoses/microbiologia , Desbridamento
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