RESUMEN
Hemolysis is the most prevalent pre-analytical interfering factor and a major source of error in laboratory analysis. The examination of samples post-centrifugation can provide valuable information regarding pre-analytical interferences. In this unusual case, a patient's plasma specimen was cherry-red after centrifugation, which is most usually indicative of hemolysis. However, subsequent investigations ruled out common hemolysis causes. We eventually determined that the patient's cherry-red plasma was most likely caused by other factors in the patient's medical history, including cancer treatment with PV-10 (rose bengal disodium 10%). We then conducted an interference study to comprehensively assess the effects of PV-10 on various biochemical tests, especially liver function tests and bilirubin levels. The findings indicate that PV-10 has varying effects on different biochemical assays and test results should be examined individually. This report underlines the need for awareness of potential drug interference on laboratory tests for better result interpretation and making clinical decisions.
Asunto(s)
Hemólisis , Humanos , Masculino , Plasma/química , Plasma/metabolismoRESUMEN
Recent studies have demonstrated the interaction between gut microbiota and brain on ischemic stroke, but the roles of gut microbiota in the pathophysiology of ischemic stroke remain largely unclear. In this study, we detected a significant increase of intestinal Akkermansia muciniphila (AKK) following ischemic stroke by a rose bengal photothrombosis model. To investigate the function and mechanism of AKK on ischemic stroke, we performed the AKK administration prior to stroke surgery. The results showed that mice treated with AKK gained significantly higher body weight and behaved better than those in PBS group at 3 days after ischemic stroke. Consistently, AKK administration remarkably decreased the infarct volumes as well as the density of degenerating neurons and apoptotic cells after ischemic stroke. Notably, AKK is a potential therapeutic target in immune-related disorders connected to the microbiota, and inflammation is crucially involved in the pathophysiological process of ischemic stroke. For the determination of underlying mechanisms of this protective effect, we investigated whether there are associations between AKK and neuroinflammation following ischemic stroke. The results suggested that AKK administration significantly reduced the activation of astrocytes and microglia but up-regulated multiple anti-inflammatory factors following ischemic stroke. Therefore, our study highlighted the beneficial roles of intestinal AKK on ischemic stroke and provided a new perspective for the treatment of ischemic stroke.
Asunto(s)
Akkermansia , Microbioma Gastrointestinal , Accidente Cerebrovascular Isquémico , Recuperación de la Función , Animales , Masculino , Ratones , Microbioma Gastrointestinal/fisiología , Ratones Endogámicos C57BL , Recuperación de la Función/fisiología , VerrucomicrobiaRESUMEN
During photosynthesis, reactive oxygen species (ROS) are formed, including hydrogen peroxide (H2O2) and singlet oxygen (1O2), which have putative roles in signalling, but their involvement in photosynthetic acclimation is unclear. Due to extreme reactivity and a short lifetime, 1O2 signalling occurs via its reaction products, such as oxidised poly-unsaturated fatty acids in thylakoid membranes. The resulting lipid peroxides decay to various aldehydes and reactive electrophile species (RES). Here, we investigated the role of ROS in the signal transduction of high light (HL), focusing on GreenCut2 genes unique to photosynthetic organisms. Using RNA seq. data, the transcriptional responses of Chlamydomonas reinhardtii to 2 h HL were compared with responses under low light to exogenous RES (acrolein; 4-hydroxynonenal), ß-cyclocitral, a ß-carotene oxidation product, as well as Rose Bengal, a 1O2-producing photosensitiser, and H2O2. HL induced significant (p < 0.05) up- and down-regulation of 108 and 23 GreenCut2 genes, respectively. Of all HL up-regulated genes, over half were also up-regulated by RES, including RBCS1 (ribulose bisphosphate carboxylase small subunit), NPQ-related PSBS1 and LHCSR1. Furthermore, 96% of the genes down-regulated by HL were also down-regulated by 1O2 or RES, including CAO1 (chlorophyllide-a oxygnease), MDH2 (NADP-malate dehydrogenase) and PGM4 (phosphoglycerate mutase) for glycolysis. In comparison, only 0-4% of HL-affected GreenCut2 genes were similarly affected by H2O2 or ß-cyclocitral. Overall, 1O2 plays a significant role in signalling during the initial acclimation of C. reinhardtii to HL by up-regulating photo-protection and carbon assimilation and down-regulating specific primary metabolic pathways. Our data support that this pathway involves RES.
Asunto(s)
Chlamydomonas reinhardtii , Fotosíntesis , Transducción de Señal , Oxígeno Singlete , Oxígeno Singlete/metabolismo , Fotosíntesis/genética , Chlamydomonas reinhardtii/genética , Chlamydomonas reinhardtii/metabolismo , Regulación de la Expresión Génica de las Plantas , Peróxido de Hidrógeno/metabolismo , Luz , Especies Reactivas de Oxígeno/metabolismoRESUMEN
PURPOSE: To develop a reliable method to generate a mouse model of branch retinal artery occlusion (BRAO) using laser-induced thrombosis of a major artery in the mouse retina. Also, to develop a reliable method to detect retinal hypoxia as predictive biomarker for the risk of neuronal cell damage in BRAO. METHODS: A reliable and reproducible model of laser-induced BRAO was developed in mouse retina using Rose Bengal. To characterize retinal hypoxia in BRAO, pimonidazole immunostaining and HYPOX-4 molecular imaging methods were used. Terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) was used to characterize neuronal cell damage in the BRAO retina. Expression of mRNA in retinal tissues from BRAO and age-matched control retinas were analyzed using qRT-PCR. RESULTS: Occlusion of a branch retinal artery near the optic nerve head (ONH) caused a pattern of retinal tissue hypoxia covering about 12.5% of the entire retina. TUNEL-positive cells were localized in all layers in BRAO retinal tissue cross sections. In addition, qRT-PCR data analysis suggests that BRAO is associated with both inflammation and hypoxia. CONCLUSIONS: This study provides a reliable method for BRAO in mouse retina and demonstrates the utility of molecular imaging method to detect retinal hypoxia as predictive biomarker for the risk of neuronal cell damage in BRAO. In addition, our data suggest that BRAO retinas are associated with inflammation and also associated with hypoxia-related neuronal cell damage. PERSPECTIVES: Imaging areas of retinal hypoxia may provide accurate diagnosis, evaluating retinal tissue injury from BRAO.
RESUMEN
Among the three active aldehyde oxidases in Arabidopsis thaliana leaves (AAO1-3), AAO3, which catalyzes the oxidation of abscisic-aldehyde to abscisic-acid, was shown recently to function as a reactive aldehyde detoxifier. Notably, aao2KO mutants exhibited less senescence symptoms and lower aldehyde accumulation, such as acrolein, benzaldehyde, and 4-hydroxyl-2-nonenal (HNE) than in wild-type leaves exposed to UV-C or Rose-Bengal. The effect of AAO2 expression absence on aldehyde detoxification by AAO3 and/or AAO1 was studied by comparing the response of wild-type plants to the response of single-functioning aao1 mutant (aao1S), aao2KO mutants, and single-functioning aao3 mutants (aao3Ss). Notably, aao3Ss exhibited similar aldehyde accumulation and chlorophyll content to aao2KO treated with UV-C or Rose-Bengal. In contrast, wild-type and aao1S exhibited higher aldehyde accumulation that resulted in lower remaining chlorophyll than in aao2KO leaves, indicating that the absence of active AAO2 enhanced AAO3 detoxification activity in aao2KO mutants. In support of this notion, employing abscisic-aldehyde as a specific substrate marker for AAO3 activity revealed enhanced AAO3 activity in aao2KO and aao3Ss leaves compared to wild-type treated with UV-C or Rose-Bengal. The similar abscisic-acid level accumulated in leaves of unstressed or stressed genotypes indicates that aldehyde detoxification by AAO3 is the cause for better stress resistance in aao2KO mutants. Employing the sulfuration process (known to activate aldehyde oxidases) in wild-type, aao2KO, and molybdenum-cofactor sulfurase (aba3-1) mutant plants revealed that the active AAO2 in WT employs sulfuration processes essential for AAO3 activity level, resulting in the lower AAO3 activity in WT than AAO3 activity in aao2KO.
RESUMEN
Healing of deep cutaneous wounds often results in detrimental sequelae, including painful and debilitating scars. Current therapies for full-thickness injuries that target specific phases of wound healing have moderate success; however, full resolution remains incomplete and negative consequences persist if skin homeostasis is not achieved. Photoactivated molecules can modulate cellular responses by generating reactive oxygen species and may provide a novel therapeutic option to improve wound healing. In the current study, we investigated the effects of Rose bengal (RB) dye in a preclinical model of full-thickness cutaneous injury. Monochromatic green light activates RB to generate ROS in the presence of oxygen, subsequently crosslinking collagen fibrils. In in vitro studies, we show that photoactivated RB is well tolerated by epidermal keratinocytes and dermal fibroblasts and can mitigate fibrotic signalling by downregulating collagen production. In a murine model of full-thickness injury, topically-applied and photoactivated RB closed wounds faster than control and vehicle treatments and showed significantly improved wound healing outcomes, including enhanced early granulation, better collagen organisation and increased vascularity in the presence of protracted tissue ROS. These data support an overall improved cutaneous wound healing profile after RB phototherapy and warrant further investigations into this versatile molecule.
RESUMEN
AIM: Impact of final root canal disinfectants diode laser (DL), Rose Bengal photosensitizer (RBP), Chitosan Nanoparticles (CNPs), and CNPs modified RBP on Enterococcus faecalis (E.faecalis) survival rate, smear layer (SL) removal efficacy and push out bond strength (PBS) of resin sealer to the radicular dentin. MATERIAL AND METHOD: Single rooted seventy-five human mandibular premolar teeth were sectioned at a level of cementoenamel (CEJ) and root canal treatment was performed using a ProTaper rotary instrument. To assess the antibacterial efficiency E.faecalis inoculation was performed using a pour plate method. The samples were arbitrarily allocated into five groups based on the final disinfectant received (n = 15) Group-1 (NaOCl+17 % EDTA), Group-2 (NaOCl+DL), Group-3 (NaOCl+CNPs), Group-4 (NaOCl+RBP activated by DL) and Group-5 (NaOCl+CNPs-RBP activated by DL). SEM analysis for SL removal was performed on five specimens from each group. Root canal obturation and sectioning were performed at cervical, middle, and apical levels. PBS and failure mode were analyzed using a universal testing machine (UTM) and stereomicroscope respectively. The data was subjected to analysis using one-way analysis of variance (ANOVA) and Tukey's post hoc test. RESULT: Group 1 (NaOCl+17 % EDTA) (0.36 ± 0.01 CFU/mL) unveiled the maximum survival rate of tested bacteria. However, Group 5 (NaOCl+CNPs-RBP activated by DL) (0.11 ± 0.01 CFU/mL) displayed minimum survival of E.faecalis. The highest SL removal (1.44 ± 0.11) and bond integrity scores (12.23 ± 0.24 MPa) were recorded in the coronal section of Group 5 (NaOCl+ CNPs-RBP activated by DL) treated roots. However, the lowest removal of SL (3.15 ± 1.09) and PBS (7.24 ± 0.09 MPa) were recorded in Group 4 (NaOCl+RBP activated by DL) treated canals. CONCLUSION: CNPs-RBP activated by DL as the final irrigant provided satisfactory antibacterial activity, SL removal, and bond strength outcomes compared to other tested groups.
RESUMEN
Previous studies have shown that antimicrobial photodynamic inactivation (aPDI) can be strongly potentiated by the addition of the non-toxic inorganic salt, potassium iodide (KI). This approach was shown to apply to many different photosensitizers, including the xanthene dye Rose Bengal (RB) excited by green light (540 nm). Rose Bengal diacetate (RBDA) is a lipophilic RB derivative that is easily taken up by cells and hydrolyzed to produce an active photosensitizer. Because KI is not taken up by microbial cells, it was of interest to see if aPDI mediated by RBDA could also be potentiated by KI. The addition of 100 mM KI strongly potentiated the killing of Gram-positive methicillin-resistant Staphylocccus aureus, Gram-negative Eschericia coli, and fungal yeast Candida albicans when treated with RBDA (up to 15 µM) for 2 hours followed by green light (540 nm, 10 J/cm2). Both RBDA aPDI regimens (400 µM RBDA with or without 400 mM KI followed by 20 J/cm2 green light) accelerated the healing of MRSA-infected excisional wounds in diabetic mice, without damaging the host tissue.
Asunto(s)
Candida albicans , Staphylococcus aureus Resistente a Meticilina , Fármacos Fotosensibilizantes , Yoduro de Potasio , Rosa Bengala , Infecciones Estafilocócicas , Cicatrización de Heridas , Animales , Rosa Bengala/farmacología , Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Cicatrización de Heridas/efectos de los fármacos , Yoduro de Potasio/farmacología , Ratones , Candida albicans/efectos de los fármacos , Fármacos Fotosensibilizantes/farmacología , Infecciones Estafilocócicas/tratamiento farmacológico , Infecciones Estafilocócicas/microbiología , Escherichia coli/efectos de los fármacos , Diabetes Mellitus Experimental/microbiología , Diabetes Mellitus Experimental/tratamiento farmacológico , Fotoquimioterapia/métodos , Sinergismo Farmacológico , Luz , MasculinoRESUMEN
BACKGROUND: Brucellosis is a zoonotic infectious disease. It is estimated that the number of cases reported today is much less than the actual number. We still have difficulty in diagnosing the disease and its organ involvement. In this sense, new approaches that can be useful in clinical practice are required, and we aimed to evaluate this situation in our study. METHODS: 171 of 213 patients followed in our center between January 2021 and April 2024 were included in the study. A total of 150 patients were included in the study as a control group. Rose Bengal test (RBT), Coombs gel test (CGT), enzyme-linked immunosorbent assay (ELISA), and automated blood culture were used for diagnosing brucellosis. Complete blood count, sedimentation, C-reactive protein, and biochemical parameters were obtained. Inflammation markers such as neutrophil-lymphocyte ratio, platelet-lymphocyte ratio, systemic immune-inflammation index, and systemic inflammation response index were calculated. RESULTS: The most successful results in the diagnosis were ELISA (89.4%), RBT (88.3%), CGT (83%), and blood culture (34.8%). For diagnosing sacroiliitis and spondylodiscitis, instead of resorting to expensive methods like magnetic resonance, a combination of ELISA positivity with elevated acute phase reactants and inflammatory markers could be significantly instructive. CONCLUSIONS: Optimizing diagnostic algorithms and exploring novel diagnostic approaches, such as inflammatory markers, hold promise for improving diagnosis and management.
RESUMEN
BACKGROUND: Brucellosis is a zoonosis endemic to specific geographical regions. In first line laboratories, diagnosis is made by blood culture or Rose Bengal (RB) serology. METHODS: We compare brucellosis testing between 2012-2021 at two university hospitals in Brussels, Belgium with concomitant national confirmed cases and institutional cases. RESULTS: RB testing increased from 30 to 211 tests/year between 2012-2021. A total of fifty-two national brucellosis cases were notified during the study period, of which fifteen cases in Brussels. No trend was noted nationally or regionally. Epidemiological data indicated travel to endemic regions, confirmed by strain testing. Institutional cases all showed symptomatic presentations with positive travel histories. CONCLUSIONS: Serologic testing inappropriately increases yearly, while annual imported brucellosis cases remain rare, and have positive travel histories and are symptomatic. We therefore support current recommendations of limiting RB testing to symptomatic patients at risk of exposure, meaning predominantly positive recent travel history.
Asunto(s)
Brucelosis , Rosa Bengala , Pruebas Serológicas , Brucelosis/diagnóstico , Brucelosis/epidemiología , Humanos , Masculino , Femenino , Bélgica/epidemiología , Adulto , Persona de Mediana Edad , Anciano , Viaje , Adulto Joven , Brucella/inmunología , Brucella/aislamiento & purificación , Hospitales Universitarios/estadística & datos numéricosRESUMEN
Photochemical sealing of a nerve wrap over the repair site isolates and optimizes the regenerating nerve microenvironment. To facilitate clinical adoption of the technology, we investigated photosealed autologous tissue in a rodent sciatic nerve transection and repair model. Rats underwent transection of the sciatic nerve with repair performed in three groups: standard microsurgical neurorrhaphy (SN) and photochemical sealing with a crosslinked human amnion (xHAM) or autologous vein. Functional recovery was assessed at four-week intervals using footprint analysis. Gastrocnemius muscle mass preservation, histology, and nerve histomorphometry were evaluated at 120 days. Nerves treated with a PTB-sealed autologous vein improved functional recovery at 120 days although the comparison between groups was not significantly different (SN: -58.4 +/- 10.9; XHAM: -57.9 +/- 8.7; Vein: -52.4 +/- 17.1). Good muscle mass preservation was observed in all groups, with no statistical differences between groups (SN: 69 +/- 7%; XHAM: 70 +/- 7%; Vein: 70 +/- 7%). Histomorphometry showed good axonal regeneration in all repair techniques. These results demonstrate that peripheral nerve repair using photosealed autologous veins produced regeneration at least equivalent to current gold-standard microsurgery. The use of autologous veins removes costs and foreign body concerns and would be readily available during surgery. This study illustrates a new repair method that could restore normal endoneurial homeostasis with minimal trauma following severe nerve injury.
Asunto(s)
Regeneración Nerviosa , Nervio Ciático , Animales , Ratas , Regeneración Nerviosa/fisiología , Nervio Ciático/lesiones , Nervio Ciático/cirugía , Nervio Ciático/fisiología , Humanos , Amnios , Trasplante Autólogo/métodos , Músculo Esquelético , Recuperación de la Función , Masculino , Procedimientos Neuroquirúrgicos/métodos , Venas/cirugíaRESUMEN
Aim: Rose Bengal photodynamic antimicrobial therapy (RB-PDAT) has poor corneal penetration, limiting its efficacy against acanthamoeba keratitis (AK). Iontophoresis enhances corneal permeation of charged molecules, piquing interest in its effects on RB in ex vivo human corneas.Methods: Five donor whole globes each underwent iontophoresis with RB, soaking in RB, or were soaked in normal saline (controls). RB penetration and corneal thickness was assessed using confocal microscopy.Results: Iontophoresis increased RB penetration compared with soaking (177 ± 9.5 µm vs. 100 ± 5.7 µm, p < 0.001), with no significant differences in corneal thickness between groups (460 ± 87 µm vs. 407 ± 69 µm, p = 0.432).Conclusion: Iontophoresis significantly improves RB penetration and its use in PDAT could offer a novel therapy for acanthamoeba keratitis. Further studies are needed to validate clinical efficacy.
The study aimed to improve a new treatment for eye infections known as photodynamic antimicrobial therapy. It investigated whether the use of electricity through a technique called iontophoresis could help a chemical called Rose Bengal go deeper into the eye in order to target more severe infections. The iontophoresis machine was custom built, with patient-contacting components 3D printed. The experiments were performed using donated human eye tissue and found that iontophoresis significantly improved the penetration depth of Rose Bengal as compared with the current technique of only soaking the eye in Rose Bengal.
Asunto(s)
Queratitis por Acanthamoeba , Córnea , Iontoforesis , Rosa Bengala , Humanos , Rosa Bengala/administración & dosificación , Iontoforesis/métodos , Córnea/metabolismo , Córnea/efectos de los fármacos , Queratitis por Acanthamoeba/tratamiento farmacológico , Fotoquimioterapia/métodos , Microscopía ConfocalRESUMEN
Assessment of the antimicrobial, micro tensile bond strength (µTBS), and degree of conversion (DC) of fifth-generation adhesive modified using photoactivated 0.5% rose bengal (RB) and photoactivated RB-doped titanium dioxide nanoparticles (TiO2NPs) in different concentrations (2% and 5%) as compared with the unmodified adhesive bonded to the carious affected dentin (CAD). Forty mandibular molars with caries progression up to the middle third of the dentin, as per the International Caries Detection and Assessment System (ICDAS) score of 4 and 5 were included. Specimens were divided into four groups based on etch and rinse adhesive (ERA) modification group 1: unmodified ERA, group 2: photoactivated 0.5% RB photosensitizer (PS) modified ERA, group 3: photoactivated RB-doped 2 wt% TiO2NPs adhesive, group 4: photoactivated RB-doped 5 wt% TiO2NPs adhesive. Followed by adhesive and composite restoration on the CAD surface. All the specimens were thermocycled and an assessment of µTBS and failure pattern analysis was performed. The antibacterial potency of RB and RB-doped TiO2NPs (2% and 5%) followed by their activation using visible light against Streptococcus mutans (S.mutans) were tested. The survival rate of S.mutans was assessed using the Kruskal-Wallis test. The analysis of µTBS involved the use of ANOVA, followed by a post-hoc Tukey honestly significant difference (HSD) multiple comparisons test. Group 1 (Unmodified ERA) (0.52 ± 0.31 CFU/mL) treated samples unveiled the highest means of bacterial survival and lowest µTBS (11.32 ± 0.63 MPa). Nevertheless, group 4: photoactivated RB-doped 5 wt% TiO2NPs adhesive displayed the lowest outcomes of S.mutans survival (0.11 ± 0.02 CFU/mL) and highest bond strength (18.76 ± 1.45 MPa). The photoactivated RB-doped 2 wt% TiO2NPs in adhesive demonstrated promising enhancements in both µTBS and antibacterial efficacy against S.mutans. However, it is noteworthy that this modification led to a decrease in the DC of the adhesive. RESEARCH HIGHLIGHTS: Unmodified ERA-treated samples unveiled the highest bacterial survival and the lowest µTBS. Photoactivated RB-doped 5 wt% TiO2NPs adhesive displayed the lowest S.mutans survival rate and highest bond strength. DC decreased with an increase in concentration of TiO2.
RESUMEN
Melanogenesis-stimulated B16-F10 cells enter in a quiescent state, present inhibited mitochondrial respiration and increased reactive oxygen species levels. These alterations suggest that these cells may be under redox signaling, allowing tumor survival. The aim of this study was to evaluate redox-modified proteins in B16-F10 cells after melanogenesis stimulation and rose bengal-photodynamic therapy (RB-PDT). A redox proteomics label-free approach based on the biotin switch assay technique with biotin-HPDP and N-ethylmaleimide was used to assess the thiol-oxidized protein profile. Aconitase was oxidized at Cys-448 and Cys-451, citrate synthase was oxidized at Cys-202 and aspartate aminotransferase (Got2) was oxidized at Cys-272 and Cys-274, exclusively after melanogenesis stimulation. After RB-PDT, only guanine nucleotide-binding protein subunit beta-2-like 1 (Gnb2l1) was oxidized (Cys-168). In contrast, melanogenesis stimulation followed by RB-PDT led to the oxidation of different cysteines in Gnb2l1 (Cys-153 and Cys-249). Besides that, glyceraldehyde-3-phosphate dehydrogenase (Gapdh) presented oxidation at Cys-245, peptidyl-prolyl cis-trans isomerase A (Ppia) was oxidized at Cys-161 and 5,6-dihydroxyindole-2-carboxylic acid oxidase (Tyrp1) was oxidized at Cys-65, Cys-30, and Cys-336 after melanogenesis stimulation followed by RB-PDT. The redox alterations observed in murine melanoma cells and identification of possible target proteins are of great importance to further understand tumor resistance mechanisms.
RESUMEN
PURPOSE: To investigate corneal biomechanical changes after corneal cross-linking (CXL) treatments with rose bengal-green light (RB-CXL) and riboflavin-UVA (RF-CXL). METHODS: A total of 60 freshly enucleated lamb eyes were obtained for this experimental study. Fifteen eyes were treated with RB-CXL using 0.1% RB solution (Group 1), 15 eyes were treated with RB-CXL using 0.2% RB solution (Group 2), 15 eyes were treated with RF-CXL using 0.1% RF solution (Group 3), and 15 eyes were used as controls (Group 4). The same treatment protocol (10-minute irradiation using a total of 5.4 J/cm2 energy) was applied to all treatment groups. To evaluate corneal biomechanical changes, the stress-strain test was used for both the treated and control corneas. The elastic modulus was calculated using the tension strain curves obtained during the test. RESULTS: The average elastic modulus values were calculated to be 18.9, 23.5, 22.3, and 14.1 MPa in Groups 1, 2, 3, and 4, respectively. Statistically significant differences were found between the groups (p < 0.001 for Group 1 vs. 2; p < 0.001 for Group 1 vs. 3; p < 0.001 for Group 1 vs. 4; p = 0.002 for Group 2 vs. 3; p < 0.001 for Group 2 vs. 4; and p < 0.001 for Group 3 vs. 4). CONCLUSIONS: In this study, the efficacy of RB-CXL treatment applied using different concentrations of RB solutions at a total energy of 5.4 J/cm2 was investigated, and 0.2% RB solution was found to have at least as much and even more effective than the RF-CXL (0.1% RF) on the corneal elasticity module. These results are encouraging for the treatment of ectatic corneas particularly below 400 µm. It is considered that the findings obtained from this study will guide future experimental and clinical studies.
Asunto(s)
Córnea , Elasticidad , Fotoquimioterapia , Fármacos Fotosensibilizantes , Riboflavina , Rosa Bengala , Rayos Ultravioleta , Animales , Fenómenos Biomecánicos , Colágeno/metabolismo , Córnea/efectos de los fármacos , Córnea/fisiopatología , Reactivos de Enlaces Cruzados , Modelos Animales de Enfermedad , Módulo de Elasticidad , Elasticidad/efectos de los fármacos , Elasticidad/fisiología , Elasticidad/efectos de la radiación , Colorantes Fluorescentes , Luz Verde , Queratocono/tratamiento farmacológico , Queratocono/fisiopatología , Fotoquimioterapia/métodos , Fármacos Fotosensibilizantes/uso terapéutico , Fármacos Fotosensibilizantes/farmacología , Riboflavina/uso terapéutico , Rosa Bengala/farmacología , OvinosRESUMEN
The inability of visible light to penetrate far through biological tissue limits its use for phototherapy and photodiagnosis of deep-tissue sites of disease. This is unfortunate because many visible dyes are excellent photosensitizers and photocatalysts that can induce a wide range of photochemical processes, including photogeneration of reactive oxygen species. One potential solution is to bring the light source closer to the site of disease by using a miniature implantable LED. With this goal in mind, we fabricated a wireless LED-based device (volume of 23 mm3) that is powered by RF energy and emits light with a wavelength of 573 nm. It has the capacity to excite the green absorbing dye Rose Bengal, which is an efficient type II photosensitizer. The wireless transfer of RF power is effective even when the device is buried in chicken breast and located 6 cm from the transmitting antenna. The combination of a wireless device as light source and Rose Bengal as photosensitizer was found to induce cell death of cultured HT-29 human colorectal adenocarcinoma cells. Time-dependent generation of protruding bubbles was observed in the photoactivated cells suggesting cell death by light-induced pyroptosis and supporting evidence was gained by cell staining with the fluorescence probes Annexin-V FITC and Propidium Iodide. The results reveal a future path towards a wireless implanted LED-based device that can trigger photodynamic immunogenic cell death in deep-seated cancerous tissue.
Asunto(s)
Fotoquimioterapia , Fármacos Fotosensibilizantes , Piroptosis , Rosa Bengala , Fármacos Fotosensibilizantes/farmacología , Piroptosis/efectos de los fármacos , Fotoquimioterapia/métodos , Humanos , Rosa Bengala/farmacología , Células HT29 , Tecnología Inalámbrica , AnimalesRESUMEN
PURPOSE: To investigate the effect of rose bengal photodynamic therapy on lipopolysaccharide-induced inflammation in human corneal fibroblasts. Furthermore, to analyze potential involvement of the mitogen-activated protein kinase and nuclear factor kappa B signaling pathways in this process. METHODS: Human corneal fibroblast cultures underwent 0-2.0 µg/mL lipopolysaccharide treatment, and 24 h later rose bengal photodynamic therapy (0.001% RB, 565 nm wavelength illumination, 0.17 J/cm2 fluence). Interleukin-6, interleukin-8, intercellular adhesion molecule-1, interferon regulatory factor-3, interferon α2, and interferon ß1 gene expressions were determined by quantitative PCR. Interleukin-6, interleukin-8, and C-C motif chemokine ligand-4 concentrations in the cell culture supernatant were measured by enzyme-linked immunosorbent assays and intercellular adhesion molecule-1 protein level in human corneal fibroblasts by western blot. In addition, the nuclear factor kappa B and mitogen-activated protein kinase signaling pathways were investigated by quantitative PCR and phosphorylation of nuclear factor kappa B p65 and p38 mitogen-activated protein kinase by western blot. RESULTS: Rose bengal photodynamic therapy in 2.0 µg/mL lipopolysaccharide-stimulated human corneal fibroblasts triggered interleukin-6 and interleukin-8 mRNA (p < .0001) and interleukin-6 protein increase (p < .0001), and downregulated intercellular adhesion molecule-1 expression (p < .001). C-C motif chemokine ligand-4, interferon regulatory factor-3, interferon α2, and interferon ß1 expressions remained unchanged (p ≥ .2). Rose bengal photodynamic therapy increased IκB kinase subunit beta, nuclear factor kappa B p65, extracellular signal-regulated kinases-2, c-Jun amino terminal kinase, and p38 transcription (p ≤ .01), and triggered nuclear factor kappa B p65 and p38 mitogen-activated protein kinase phosphorylation (p ≤ .04) in lipopolysaccharide treated human corneal fibroblasts. CONCLUSION: Rose bengal photodynamic therapy of lipopolysaccharide-stimulated human corneal fibroblasts can modify the inflammatory response by inducing interleukin-6 and interleukin-8 expression, and decreasing intercellular adhesion molecule-1 production. C-C motif chemokine ligand-4, interferon regulatory factor-3, and interferon α and ß expressions are not affected by rose bengal photodynamic therapy in these cells. The underlying mechanisms may be associated with nuclear factor kappa B and p38 mitogen-activated protein kinase pathway activation.
Asunto(s)
Ensayo de Inmunoadsorción Enzimática , Lipopolisacáridos , FN-kappa B , Fotoquimioterapia , Rosa Bengala , Transducción de Señal , Proteínas Quinasas p38 Activadas por Mitógenos , Humanos , Rosa Bengala/farmacología , Fotoquimioterapia/métodos , Lipopolisacáridos/farmacología , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo , Células Cultivadas , FN-kappa B/metabolismo , Western Blotting , Fármacos Fotosensibilizantes/farmacología , Queratocitos de la Córnea/metabolismo , Queratocitos de la Córnea/efectos de los fármacos , Fibroblastos/metabolismo , Fibroblastos/efectos de los fármacos , Regulación de la Expresión Génica , Reacción en Cadena en Tiempo Real de la Polimerasa , Inflamación/metabolismo , Inflamación/tratamiento farmacológico , Factor 3 Regulador del Interferón/metabolismo , Molécula 1 de Adhesión Intercelular/metabolismo , Molécula 1 de Adhesión Intercelular/genéticaRESUMEN
Bisphenol A (BPA) is a widespread organic micro-pollutant, found in most environments, including alpine and Arctic regions, and several matrices such as waters and aerosols. Polar regions are characterized by periods of intense irradiation with no sunset due to the continuous sunlight, while alpine areas, despite following the day-night cycle of mid-latitudes, also undergo strong irradiation. For such conditions, it is possible that a fraction of the BPA present in snow may degrade through direct photolysis, producing other unknown species with different environmental mobility and possible ecotoxic effects. Furthermore, the snowpack is rich in species (known as photosensitizers) that facilitate indirect photodegradation processes through reactions involving hydroxyl radicals · OH , singlet oxygen (1O2), excited triplet states of the organic fraction (3CDOM*), and nitrite/nitrate. In this study, we investigated both direct and indirect photodegradation of BPA in the presence of specific photosensitizers producing · OH , 1O2, 3CDOM*, and NO2- to specifically explore the products of the reaction. The study was conducted in both liquid water and ice, under light and dark conditions. Results, obtained by HPLC-HRMS, revealed that the matrix in which the reaction takes place, in addition to the photosensitizer used, may influence the degradation by-products. This allows for the possibility of distinguishing the reaction environment based on the identified product.
RESUMEN
PV-10 is a 10% formulation of rose bengal sodium that has potent immunotherapeutic and anti-cancer activity against various tumors, including metastatic melanoma and refractory neuroblastoma. Currently, PV-10 is undergoing clinical testing for refractory metastatic neuroendocrine cancer and melanomas. However, preclinical investigation of PV-10 activity and its mechanisms against phenotypically and molecularly diverse adult solid tumors had not been conducted. In a panel of human cell lines derived from breast, colorectal, head and neck, and testicular cancers, we demonstrated that PV-10 induces cytotoxicity by apoptotic and autophagic pathways involving caspase-mediated PARP cleavage, downregulation of SQSTM1/p62, and upregulation of beclin-1. Treatment with PV-10 also consistently reduced phosphorylation of WNK1, which has been implicated in cancer cell migration and autophagy inhibition. By wound healing assay, PV-10 treatment inhibited the migration of cancer cells. Finally, significant inhibition of tumor growth was also noted in tumor-bearing mice treated with PV-10 by intralesional or systemic administration. In addition to known PV-10-mediated tumor-specific cytotoxic effects, we identified the mechanisms of PV-10 and provide new insights into its effect on autophagy and metastasis. Our data provide essential mechanism-based evidence and biomarkers of activity to formulate clinical studies of PV-10 in the future.
RESUMEN
Dental diseases, including conditions affecting oral structures, have become more common due to unhealthy lifestyle choices. Traditional antibiotic treatments face challenges related to the development of antibiotic resistance in bacteria. Photodynamic antibacterial chemotherapy is emerging as a promising alternative using photosensitizers to generate reactive oxygen species upon exposure to light. This article examines the photosensitizer Rose Bengal (RB) immobilized in hyaluronic acid (HA) for prolonged antibacterial action. The RB-HA conjugate demonstrated a molar ratio of approximately three RB residues to each of the ten units of HA. RB-HA exhibited a high singlet oxygen quantum yield (ΔΦ = 0.90), suggesting its efficacy in photodynamic treatment. A photostability analysis revealed slower photobleaching of RB-HA, which is essential for prolonged application. Under visible light and ultrasonic treatment, RB-HA exhibited effective antibacterial activity against Gram-positive S. aureus and Gram-negative E. coli bacteria for at least 80 days. The gradual release of RB ensured sustained bactericidal concentration. The study establishes RB-HA as a promising candidate for antimicrobial photodynamic and sonodynamic therapy in dental and other medical fields, providing enhanced stability and prolonged antibacterial efficacy.