RESUMEN
This study aimed to evaluate a novel albumin-binding strategy for addressing the challenge of insufficient tumor retention of fibroblast activation protein inhibitors (FAPIs). Maleimide, a molecule capable of covalent binding to free thiol groups, was modified to conjugate with FAPI-04 in order to enhance its binding to endogenous albumin, resulting in an extended blood circulation half-life and increased tumor uptake. DOTA-FAPI-maleimide was prepared and radiolabeled with Ga-68 and Lu-177, followed by cellular assays, pharmacokinetic analysis, PET/CT, and SPECT/CT imaging to assess the probe distribution in various tumor-bearing models. Radiolabeling of the modified probe was successfully achieved with a radiochemical yield of over 99% and remained stable for 144 h. Cellular assays showed that the ligand concentration required for 50% inhibition of the probe was 1.20 ± 0.31 nM, and the Kd was 0.70 ± 0.07 nM with a Bmax of 7.94 ± 0.16 fmol/cell, indicative of higher specificity and affinity of DOTA-FAPI-maleimide compared to other FAPI-04 variants. In addition, DOTA-FAPI-maleimide exhibited a persistent blood clearance half-life of 7.11 ± 0.34 h. PET/CT images showed a tumor uptake of 2.20 ± 0.44%ID/g at 0.5 h p.i., with a tumor/muscle ratio of 5.64 in HT-1080-FAP tumor-bearing models. SPECT/CT images demonstrated long-lasting tumor retention. At 24 h p.i., the tumor uptake of [177Lu]Lu-DOTA-FAPI-maleimide reached 5.04 ± 1.67%ID/g, with stable tumor retention of 3.40 ± 1.95%ID/g after 4 days p.i. In conclusion, we developed and evaluated the thiol group-attaching strategy, which significantly extended the circulation and tumor retention of the adapted FAPI tracer. We envision its potential application for clinical cancer theranostics.
Asunto(s)
Maleimidas , Tomografía Computarizada por Tomografía de Emisión de Positrones , Radiofármacos , Animales , Maleimidas/química , Ratones , Humanos , Radiofármacos/farmacocinética , Radiofármacos/química , Distribución Tisular , Línea Celular Tumoral , Tomografía Computarizada por Tomografía de Emisión de Positrones/métodos , Radioisótopos de Galio/farmacocinética , Radioisótopos de Galio/química , Radioisótopos/química , Compuestos Heterocíclicos con 1 Anillo/química , Compuestos Heterocíclicos con 1 Anillo/farmacocinética , Femenino , Neoplasias/diagnóstico por imagen , Neoplasias/tratamiento farmacológico , Ratones Desnudos , Tomografía Computarizada por Tomografía Computarizada de Emisión de Fotón Único/métodos , Ensayos Antitumor por Modelo de Xenoinjerto , Endopeptidasas , Proteínas de la Membrana/metabolismo , Nanomedicina Teranóstica/métodos , LutecioRESUMEN
The stimulator of interferon genes (STING) is a pivotal protein in the production of STING-dependent type I interferon, which has the potential to enhance tumor rejection. The visualization of STING in the tumor microenvironment is valuable for STING-related treatments, but few STING imaging probes have been reported to date. In this study, we developed a novel 18F-labeled agent ([18F]F-CRI1) with an acridone core structure for the positron emission tomography (PET) imaging of STING in CT26 tumors. The probe was successfully prepared with a nanomolar STING binding affinity of Kd = 40.62 nM. [18F]F-CRI1 accumulated quickly in the tumor sites and its uptake reached a maximum of 3.02 ± 0.42% ID/g after 1 h i.v. injection. The specificity of [18F]F-CRI1 was confirmed both in in vitro cell uptake and in vivo PET imaging by blocking studies. Our findings suggest that [18F]F-CRI1 may be a potential agent for visualizing STING in the tumor microenvironment.
Asunto(s)
Radioisótopos de Flúor , Neoplasias , Humanos , Tomografía de Emisión de Positrones/métodos , Neoplasias/diagnóstico por imagen , Interferones , Línea Celular Tumoral , Microambiente TumoralRESUMEN
Pancreatic cancer (PC) is one of the most common malignant tumors in digestive tract. To explore the role of epigenetic factor EZH2 in the malignant proliferation of PC, so as to provide effective medical help in PC. Sixty paraffin sections of PC were collected and the expression of EZH2 in PC tissues was detected by immunohistochemical assay. Three normal pancreas tissue samples were used as controls. The regulation of EZH2 gene on proliferation and migration of normal pancreatic cell and PC cell were determined by MTS, colony forming, Ki-67 antibody, scratch and Transwell assays. Through differential gene annotation and differential gene signaling pathway analysis, differentially expressed genes related to cell proliferation were selected and verified by RT-qPCR. EZH2 is mainly expressed in the nuclei of pancreatic tumor cells, but not in normal pancreatic cells. The results of cell function experiments showed that EZH2 overexpression could enhance the proliferation and migration ability of PC cell BXPC-3. Cell proliferation ability increased by 38% compared to the control group. EZH2 knockdown resulted in reduced proliferation and migration ability of cells. Compared with control, proliferation ability of cells reduced by 16%-40%. The results of bioinformatics analysis of transcriptome data and RT-qPCR demonstrated that EZH2 could regulate the expression of E2F1, GLI1, CDK3 and Mcm4 in normal and PC cells. The results revealed that EZH2 might regulate the proliferation of normal pancreatic cell and PC cell through E2F1, GLI1, CDK3 and Mcm4.
Asunto(s)
Neoplasias Pancreáticas , Humanos , Proteína con Dedos de Zinc GLI1/metabolismo , Línea Celular Tumoral , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/metabolismo , Neoplasias Pancreáticas/patología , Páncreas/metabolismo , Páncreas/patología , Componente 4 del Complejo de Mantenimiento de Minicromosoma/metabolismo , Proteína Potenciadora del Homólogo Zeste 2/genética , Proteína Potenciadora del Homólogo Zeste 2/metabolismo , Quinasa 3 Dependiente de Ciclina/metabolismo , Factor de Transcripción E2F1/metabolismo , Neoplasias PancreáticasRESUMEN
PURPOSE: Stimulator of interferon genes (STING) protein plays a vital role in the immune surveillance of tumor microenvironment. Monitoring STING expression in tumors benefits the relevant STING therapy. This study aimed to develop a novel 18F-labeled agonist, dimeric amidobenzimidazole (diABZI), and firstly evaluate the feasibility of noninvasive positron emission tomography (PET) imaging of STING expression in the tumor microenvironment. METHODS: An analog of the STING agonist NOTA-DABI was synthesized and labeled with 18F via Al18F-NOTA complexation (denoted as [18F]F-DABI). Physicochemical properties, STING protein-binding affinity, and specificity of [18F]F-DABI were evaluated using cell uptake and docking assays. In vivo small-animal PET imaging and biodistribution studies of [18F]F-DABI in tumor-bearing mice were performed to verify the pharmacokinetics and tumor targeting ability. The correlation between tumor uptake and STING expression was also analyzed. RESULTS: [18F]F-DABI was produced conveniently with high radiochemical yield (44 ± 15%), radiochemical purity (> 97%) and molar activity (15-30 GBq/µmol). In vitro binding assays demonstrated that [18F]F-DABI has a favorable affinity and specificity for STING with a KD of 12.98 ± 2.07 nM. In vivo studies demonstrated the specificity of [18F]F-DABI for PET imaging of STING expression with B16F10 tumor uptake of 10.93 ± 0.93%ID/g, which was significantly different from that of blocking groups (3.13 ± 0.88%ID/g, ***p < 0.0001). Furthermore, tumor uptake of [18F]F-DABI was well positively correlated with STING expression in different tumor types. Biodistribution results demonstrated that [18F]F-DABI was predominately uptaken in the liver and intestines, indicating its hepatobiliary elimination. CONCLUSION: This proof-of-concept study demonstrated a STING-binding radioligand for PET imaging, which could be used as a potential companion diagnostic tool for related STING-agonist therapies.
Asunto(s)
Radioisótopos de Flúor , Tomografía de Emisión de Positrones , Animales , Ratones , Radioisótopos de Flúor/farmacocinética , Distribución Tisular , Línea Celular Tumoral , Tomografía de Emisión de Positrones/métodos , Expresión Génica , InterferonesRESUMEN
PURPOSE: Bacterial infection and antibiotic resistance are serious threats to human health. This study aimed to develop two novel radiotracers, 18F-NTRP and 18F-NCRP, that possess a specific nitroreductase (NTR) response to image deep-seated bacterial infections using positron emission tomography (PET). This method can distinguish infection from sterile inflammation. METHODS: 18F-NTRP and 18F-NCRP were synthesized via a one-step method; all the steps usually involved in tracer radiosynthesis were successfully adapted in the All-In-One automated module. After the physiochemical properties of 18F-NTRP and 18F-NCRP were characterized, their specificity and selectivity for NTR were verified in E. coli and S. aureus. The ex vivo biodistribution of the tracers was evaluated in normal mice. MicroPET-CT imaging was performed in mouse models of bacterial infection and inflammation after the administration of 18F-NTRP or 18F-NCRP. RESULTS: Fully automated radiosynthesis of 18F-NTRP and 18F-NCRP was achieved within 90-110 min with overall decay-uncorrected, isolated radiochemical yields of 21.24 ± 4.25% and 11.3 ± 3.78%, respectively. The molar activities of 18F-NTRP and 18F-NCRP were 320 ± 40 GBq/µmol and 275 ± 33 GBq/µmol, respectively. In addition, 18F-NTRP and 18F-NCRP exhibited high selectivity and specificity for NTR response. PET-CT imaging in bacteria-infected mouse models with 18F-NTRP or 18F-NCRP showed significant radioactivity uptake in either E. coli- or S. aureus-infected muscles. The uptake for E. coli-infected muscles, 2.4 ± 0.2%ID/g with 18F-NTRP and 4.05 ± 0.49%ID/g with 18F-NCRP, was up to three times greater than that for uninfected control muscles. Furthermore, for both 18F-NTRP and 18F-NCRP, the uptake in bacterial infection was 2.6 times higher than that in sterile inflammation, allowing an effective distinction of infection from inflammation. CONCLUSION: 18F-NTRP and 18F-NCRP are worth further investigation to verify their potential clinical application for distinguishing bacterial infection from sterile inflammation via their specific NTR responsiveness.
Asunto(s)
Infecciones Bacterianas , Mecloretamina , Animales , Escherichia coli , Radioisótopos de Flúor/química , Humanos , Inflamación/diagnóstico por imagen , Ratones , Nitrorreductasas , Tomografía Computarizada por Tomografía de Emisión de Positrones , Tomografía de Emisión de Positrones/métodos , Staphylococcus aureus , Distribución Tisular , Tomografía Computarizada por Rayos XRESUMEN
OBJECTIVES: To analyse the clinical features and risk factors of acute/subacute interstitial pneumonia (A/SIP) and death in patients with positive anti-aminoacyl-transfer RNA synthetase antibody (anti-ARS Ab) and positive anti-melanoma differentiation-associated gene 5 antibodies (anti-MDA5 Ab). METHODS: Interstitial lung disease (ILD) patients with anti-ARS+ or anti-MDA5+ were recruited. Their demographics, clinical manifestations, laboratory data were collected and they were followed up for 1 year. Risk factors of A/SIP and mortality were analysed. RESULTS: 71 patients with anti-ARS+ ILD and 31 patients with anti-MDA5+ ILD were included. Incidence of ulcerative rash, Gottron's sign, pulmonary infection and A/SIP in the anti-MDA5+ group were significantly higher than those in the anti-ARS+ group, Creatine kinase (CK), leukocyte count, and lymphocyte count were lower, the value of serum ferritin (SF) was higher, and 12-month cumulative survival rate was lower. Advanced age, anti-MDA5+ and low immunoglobulin G (IgG) level were independent predictors of A/SIP. The decreased PaO2 and elevated SF were independent predictors for poor prognosis in A/SIP patients. CONCLUSIONS: Compared to anti-ARS+ group, the anti-MDA5+ group was more prone to ulcerative rash, Gottron's sign and pulmonary infection. Patients with anti-MDA5+, advanced age and decreased values of IgG were more likely to have A/SIP, while patients with A/SIP had lower incidence of myositis and arthritis. Mortality of A/SIP patients increased with higher serum ferritin level.
Asunto(s)
Aminoacil-ARNt Sintetasas , Dermatomiositis , Enfermedades Pulmonares Intersticiales , Autoanticuerpos , Humanos , Helicasa Inducida por Interferón IFIH1 , Enfermedades Pulmonares Intersticiales/diagnóstico , Pronóstico , Estudios RetrospectivosRESUMEN
Phthalic acid esters (PAEs) can enter environment media by secondary effluent discharge from wastewater treatment plants (WWTP) into receiving rivers, thus posing a threat to ecosystem health. A level III fugacity model was established to simulate the fate and transfer of four PAEs in a study area in Tianjin, China, and to evaluate the influence of WWTP discharge on PAEs levels in the receiving river. The results show that the logarithmic residuals of most simulated and measured values of PAEs are within one order of magnitude with a good agreement. PAEs in the study area were mainly distributed in soil and sediment phases, which accounted for 84.66%, 50.26%, 71.96% and 99.09% for dimethyl phthalate (DMP), diethyl phthalate (DEP), dibutyl phthalate (DBP) and di-(2-ethylhexyl) phthalate (DEHP), respectively. The upstream advection accounted for 77.90%, 93.20%, 90.21% and 90.93% of the total source of DMP, DEP, DBP and DEHP in the river water, respectively, while the contribution of secondary effluent discharge was much lower. Sensitivity analysis shows that emission and inflow parameters have greater influences on the multimedia distributions of PAEs than physicochemical and environmental parameters. Monte Carlo analysis quantifies the uncertainties and verifies the reliability of the simulation results.
Asunto(s)
Dietilhexil Ftalato , Ácidos Ftálicos , Contaminantes Químicos del Agua , Purificación del Agua , China , Dibutil Ftalato , Dietilhexil Ftalato/análisis , Ecosistema , Ésteres/análisis , Multimedia , Ácidos Ftálicos/análisis , Reproducibilidad de los Resultados , Ríos , Contaminantes Químicos del Agua/análisisRESUMEN
BACKGROUND: Identifying gene mutation signatures will enable a better understanding for the occurrence and development of colorectal cancer (CRC), and provide some potential biomarkers for clinical practice. Currently, however, there is still few effective biomarkers for early diagnosis and prognostic judgment in CRC patients. The purpose was to identify novel mutation signatures for the diagnosis and prognosis of CRC. METHODS: Clinical information of 531 CRC patients and their sequencing data were downloaded from TCGA database (training group), and 53 clinical patients were collected and sequenced with targeted next generation sequencing (NGS) technology (validation group). The relationship between the mutation genes and the diagnosis, pathological type, stage and prognosis of CRC were compared to construct signatures for CRC, and then analyzed their relationship with RNA expression, immunocyte infiltration and tumor microenvironment (TME). RESULTS: Mutations of TP53, APC, KRAS, BRAF and ATM covered 97.55% of TCGA population and 83.02% validation patients. Moreover, 57.14% validation samples and 22.06% TCGA samples indicated that patients with mucinous adenocarcinoma tended to have BRAF mutation, but no TP53 mutation. Mutations of TP53, PIK3CA, FAT4, FMN2 and TRRAP had a remarkable difference between I-II and III-IV stage patients (P < 0.0001). Besides, the combination of PIK3CA, LRP1B, FAT4 and ROS1 formed signatures for the prognosis and survival of CRC patients. The mutations of TP53, APC, KRAS, BRAF, ATM, PIK3CA, FAT4, FMN2, TRRAP, LRP1B, and ROS1 formed the signatures for predicting diagnosis and prognosis of CRC. Among them, mutation of TP53, APC, KRAS, BRAF, ATM, PIK3CA, FAT4 and TRRAP significantly reduced their RNA expression level. Stromal score, immune score and ESTIMATE score were lower in patients with TP53, APC, KRAS, PIK3CA mutation compared non-mutation patients. All the 11 gene mutations affected the distributions of immune cells. CONCLUSION: This study constructed gene mutation signatures for the diagnosis, treatment and prognosis in CRC, and proved that their mutations affected RNA expression levels, TME and immunocyte infiltration. Our results put forward further insights into the genotype of CRC.
Asunto(s)
Biomarcadores de Tumor , Neoplasias Colorrectales/genética , Mutación , Adulto , Anciano , Alelos , Neoplasias Colorrectales/diagnóstico , Neoplasias Colorrectales/inmunología , Neoplasias Colorrectales/mortalidad , Biología Computacional/métodos , Femenino , Estudios de Asociación Genética , Genotipo , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Masculino , Persona de Mediana Edad , Clasificación del Tumor , Estadificación de Neoplasias , Pronóstico , Proteínas Proto-Oncogénicas/genética , Análisis de Supervivencia , Microambiente Tumoral/genética , Microambiente Tumoral/inmunología , Proteínas Supresoras de Tumor/genéticaRESUMEN
In recent years, α-glucosidase inhibitors (AGIs) have played a significant role in the treatment of type II diabetes (T2D), so it is necessary to develop a reliable and sensitive method to find new AGIs. Herein, we establish a novel method based on fluorescent carbon nitride nanoparticles (CNNPs) for the sensitive detection of the activity of α-glucosidase (α-glu) and the screening of its inhibitors. A CNNP-based fluorescent probe is synthesized from green raw materials, urea and lysine, by a one-pot method. In the presence of α-glu, the substrate 4-nitrophenyl-α-d-glucopyranoside (pNPG) is hydrolyzed to generate 4-nitrophenol (pNP), leading to the fluorescence (FL) quenching of CNNPs due to the inner filter effect (IFE). On the other hand, the activity of α-glu is inhibited after the addition of AGIs, which turns on the FL of CNNPs. In this way, the detection of α-glu activity and the screening of AGIs are achieved. The linear range is 1.25-10.00 U L-1 with a limit of detection as low as 0.17 U L-1 and the IC50 values of two typical inhibitors (gallic acid and acarbose) are 813 µM and 465 µM, respectively. The CNNP probe is further applied for the determination of α-glu activity in human serum samples with satisfactory results.
Asunto(s)
Diabetes Mellitus Tipo 2 , Nanopartículas , Carbono , Diabetes Mellitus Tipo 2/diagnóstico , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Colorantes Fluorescentes , Humanos , Nitrilos , alfa-GlucosidasasRESUMEN
Amblyopia is a relatively common (incidence 3%) developmental disorder in which there is loss of vision as a consequence of a disruption to normal visual development. Although the deficit is monocular and known to be of cortical origin, the nature of the processing deficit is controversial. Human behavioral studies have identified two main deficits - a loss of contrast sensitivity and perceived spatial distortions. Here we use a multifocal fMRI approach to ascertain, in a group of anisometropic amblyopes, whether these two deficits have a single common cause or whether they are the result of two underlying independent cortical disorders. We found that fMRI magnitudes were attenuated in amblyopic eye stimulation, and that there was poor fidelity for co-localization of the activity clusters between the amblyopic and fellow-fixing eye stimulation. These effects varied across eccentricities and correlate with the degree of amblyopia but not with one another, suggesting two independent cortical deficits: a reduced responsiveness as well as reduced fidelity of spatial representation. These deficits are independent of eccentricity within the central field and consistent across early cortical visual areas.
Asunto(s)
Ambliopía/fisiopatología , Mapeo Encefálico/métodos , Sensibilidad de Contraste/fisiología , Reconocimiento Visual de Modelos/fisiología , Trastornos de la Percepción/fisiopatología , Distorsión de la Percepción/fisiología , Percepción Espacial/fisiología , Corteza Visual/fisiopatología , Adulto , Ambliopía/diagnóstico por imagen , Femenino , Humanos , Imagen por Resonancia Magnética , Masculino , Trastornos de la Percepción/diagnóstico por imagen , Corteza Visual/diagnóstico por imagen , Adulto JovenRESUMEN
Novel immune checkpoint blockades, including those targeting CD73 and A2aR, are being evaluated in malignancies in clinical trials. Here, we investigated the expression of CD73 and A2aR as well as tumor-infiltrating lymphocytes (TILs), and analyzed their correlations with clinicopathological characteristics and survival in diffuse large B-cell lymphoma (DLBCL). We found that CD73 expression on tumor cells, rather than the total protein and gene levels of CD73, was associated with survival. Patients with CD73+ /Pax-5+ (median survival, 57.8 months; 95% CI, 46.4-69.3) experienced significantly poorer outcomes than those with CD73- /Pax-5+ (median survival, 73.5 months; 95% CI, 65.9-81.2). Additionally, A2aR expression on both total TILs and CD8+ TILs was correlated with survival. Patients with A2aR+ TILs (median survival, 53.3 months; 95% CI, 40.6-66.0) had a significantly shorter survival time than patients with A2aR- TILs (median survival, 74.5 months; 95% CI, 67.5-81.5). Spearman's rank test showed that CD73 expression on tumor cells was positively correlated with A2aR expression on TILs (R = 0.395, p = 0.001). We further found that patients could be more precisely stratified through the combination of CD73 tumor cell expression and A2aR TILs expression, and patients with CD73+ /Pax-5+ and A2aR+ TILs experienced the worst outcome. We also revealed that patients with CD73+ /Pax-5+ and low CD8+ TILs or low absolute lymphocyte counts had unfavorable outcomes. Overall, our findings uncovered that patients with CD73+ on tumor cells as well as A2aR+ on TILs or low CD8+ TILs exhibited inferior survival, supporting potential combination strategies using CD73/A2aR immunosuppressive blockades as treatment options for DLBCL patients.
Asunto(s)
5'-Nucleotidasa/inmunología , Linfocitos Infiltrantes de Tumor/inmunología , Linfoma de Células B Grandes Difuso/inmunología , Receptor de Adenosina A2A/inmunología , 5'-Nucleotidasa/biosíntesis , Protocolos de Quimioterapia Combinada Antineoplásica/administración & dosificación , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Biomarcadores de Tumor/biosíntesis , Biomarcadores de Tumor/inmunología , Linfocitos T CD8-positivos/inmunología , Ciclofosfamida/administración & dosificación , Doxorrubicina/administración & dosificación , Femenino , Proteínas Ligadas a GPI/biosíntesis , Proteínas Ligadas a GPI/inmunología , Humanos , Linfoma de Células B Grandes Difuso/tratamiento farmacológico , Linfoma de Células B Grandes Difuso/patología , Masculino , Persona de Mediana Edad , Factor de Transcripción PAX5/biosíntesis , Factor de Transcripción PAX5/inmunología , Prednisona/administración & dosificación , Receptor de Adenosina A2A/biosíntesis , Rituximab/administración & dosificación , Transducción de Señal/inmunología , Tasa de Supervivencia , Vincristina/administración & dosificaciónRESUMEN
Photolysis and photoracemization are two important photochemical phenomena of the prototype complexes [Ru(bipy)3]2+ and [Ru(phen)3]2+ (bipy = 2,2'-bipyridine, phen = 1,10-phenanthroline), but little is known about their relations. To solve this issue, the photoinduced chiral inversion ΔâΛ of the complexes was analyzed theoretically. The results indicated that the photoracemization reaction proceeds on the lowest triplet potential energy surface in three steps 3CTΔâ3MCΔ, 3MCΔâ3MCΛ, and 3MCΛâ3CTΛ (CT = charge transfer state; MC = metal-centered state). Where the first and third steps are fast processes of picoseconds, the second is the rate-determining step (RDS) of microseconds. Such a slow step for the racemization leads to the excited molecule lingering around the bottom of 3MC state after the first step and, therefore, greatly enhances the possibility of deexcitation and photolysis mostly at the triplet-singlet crossing point. In other words, the photoracemization and photolysis of the complexes have a competition relation, not a slave relation as assumed by the photoracemization model suggested in literature. They are dominated by the RDS. This conclusion is also consistent with the Δ(δ S)âΛ(δ S) chiral inversion of the [Ru(bipy)2(L-ser)]+ series complexes, which is reversible with no detectable photolysis, as its second step is a fast one. Note that, although the photoracemization of the prototype complexes is very slow, it passes through the three steps reversibly and ends with a photon emitting, which could be detected with the time-resolved circularly polarized luminescence and related techniques. These findings are helpful to understand and control the photochemical behavior of the complexes in practice.
RESUMEN
In this study, we investigated the effect of optical treatment on sensory eye balance in anisometropic amblyopia. Fourteen individuals (age: 13.7 ± 8.4 years old) with previously untreated anisometropic amblyopia were enrolled in the study. The average magnitude of their anisometropia (spherical equivalent) was 4.02 ± 1.19 DS. Their best corrected monocular visual acuity and sensory eye balance were measured before and after a 2-month period of full refractive correction (i.e., our optical treatment). Spectacle-corrected distance visual acuity (at 5 m) was measured monocularly using the Tumbling E Chart. Sensory eye balance was quantitatively assessed using a binocular phase-combination paradigm to determine the interocular contrast ratio at which the two eyes were balanced in binocular sensory combination (i.e., the balance point). We found that both interocular contrast ratio at the balance point (p = 0.006) and visual acuity of the amblyopic eye (p < 0.001) were significantly improved after 2 months of optical treatment, often referred to as refractive adaptation. We conclude that sustained optical treatment improves interocular sensory balance in anisometropic amblyopia as well as monocular acuity. Optical treatment is a passive form of binocular therapy and a necessary first step in treating the binocular dysfunction that characterizes amblyopia.
Asunto(s)
Ambliopía/terapia , Anisometropía/terapia , Anteojos , Visión Binocular/fisiología , Adaptación Fisiológica , Adolescente , Adulto , Ambliopía/fisiopatología , Anisometropía/fisiopatología , Niño , Femenino , Humanos , Masculino , Retinoscopía , Agudeza Visual/fisiología , Adulto JovenRESUMEN
One of the main concerns in the photochemistry and photophysics of ruthenium complexes is the de-excitation of the triplet metal centered ligand-field state 3MC. To understand the mechanism by which the 3MC states in some reversible photochemical reactions could avoid the fate of fast decay and ligand dissociations, the photoinduced chiral inversion at the metal center of the complexes [Ru(diimine)2(l-ser)]+ (diimine = 1,10-phenanthroline or 2,2'-bipyridine, l-ser = l-serine) has been analyzed at the first principle level of theory. The calculated equilibrium constants and ECD curves for the photoinduced equilibrium mixtures are in agreement with the observed ones. The results showed that the reversible photochemical process Δ(δS) â Λ(δS) on the potential surface of the lowest triplet excited state proceeds in three steps: 3CTΔ â 3MCΔ, 3MCΔ â 3MCΛ, 3MCΛ â 3CTΛ, where the first and the third steps involve mainly the elongation and compression of the octahedral core of the reactant Δ(δS) and product Λ(δS), respectively. The chiral inversion Δ â Λ takes place in the second step through a much distorted square-pyramid-like transition state, and actually proceeds on the triplet ground state 3MC due to the crossover of the triplet T1 and singlet S0 states. Inspecting the transient structures at the crossing points, we found that they become less distorted and their lowest or imaginary-frequency displacement vectors in triplet state still dominate the reaction path, which makes the reaction reversible without ligand release. Thus, the triplet ground-state-bridged photoinduced mechanism offers a new angle of view to understand the related reversible photochemical reactions.
RESUMEN
Amblyopia results from inadequate visual experience during the critical period of visual development. Abnormal binocular interactions are believed to play a critical role in amblyopia. These binocular deficits can often be resolved, owing to the residual visual plasticity in amblyopes. In this study, we quantitatively measured the sensory eye dominance in treated anisometropic amblyopes to determine whether they had fully recovered. Fourteen treated anisometropic amblyopes with normal or corrected to normal visual acuity participated, and their sensory eye dominance was assessed by using a binocular phase combination paradigm. We found that the two eyes were unequal in binocular combination in most (11 out of 14) of our treated anisometropic amblyopes, but none of the controls. We concluded that the treated anisometropic amblyopes, even those with a normal range of visual acuity, exhibited abnormal binocular processing. Our results thus suggest that there is potential for improvement in treated anisometropic amblyopes that may further enhance their binocular visual functioning.
Asunto(s)
Ambliopía/fisiopatología , Predominio Ocular , Visión Binocular , Agudeza Visual , Ambliopía/rehabilitación , Niño , Percepción de Profundidad , Femenino , Humanos , Masculino , Plasticidad Neuronal , Recuperación de la Función , Resultado del TratamientoRESUMEN
Compared with the importance of didentate ligands to chiral chelates, little is known about the influence of unidentate ligands on the chiroptical properties of related chelates. To assess the importance of achiral unidentate ligands to electronic circular dichroism (ECD) spectra, calculations of the excitation energies and oscillator and rotational strengths for all the Λ-enantiomers of bis(ethylenediamine) cobalt(III) complexes cis-[Co(en)2(X)2](n+) (X = Cl(-), CN(-), NH3, N3(-), NO2(-), H2O; n = 1, 3) were performed at the TDDFT/B3LYP/6-311++G(2d,p)//DFT/B3LYP/6-311++G(2d,p) level of theory, including solvent effects. The individual contributions of the chiral arrays, Δ/Λ octahedral core, δ/λ twists of the en ligands, and δ/λ relative orientations of the unidentate ligands to the rotational strengths of related transitions were quantitatively determined and graphically presented. It was found that, for the chelates with nonaxially symmetric unidentate ligands (N3(-), NO2(-), H2O), the chiral orientation (δ/λ) of the unidentate ligands not only dominates the ECD spectra of the Λ-diastereoisomers but also dominates the relative energies in solution with the δ-orientation preferred. For those complexes with axially symmetric unidentate ligands (Cl(-), CN(-), NH3), the inherent dissymmetry within the metal ion-donor atom cluster was found to be unidentate ligand-dependent. The Boltzmann-weighted averaged ECD spectra of the complexes with Λ-octahedral core are in excellent agreement with the observed ones, except that for the diazido complex, they are opposite in sign. This demonstrates that the absolute configuration of the complex cis-(-)-[Co(en)2(N3)2](+) is Δ, not the Λ-form assigned by McCaffery et al. in 1965. These findings not only reveal the importance of unidentate ligands to the ECD spectra of related chelates but also provide an insight for the influence of unidentate ligands on both the inherent dissymmetry and the distributional chirality.
RESUMEN
Bacillus cereus is responsible for several outbreaks of foodborne diseases due to its emetic toxin and enterotoxin. Enterotoxins, cytotoxin K (CytK), nonhemolytic enterotoxin (Nhe), and hemolysin BL (Hbl), have been recorded in several diarrheal cases due to food poisoning from B. cereus. The objective of this study was to develop a rapid and accurate method that combines multiplex PCR with propidium monoazide to selectively detect viable cells of enterotoxin-producing B. cereus in milk powder, noodles, and rice, and investigate the distribution of enterotoxins in 62 strains of B. cereus in Jiangxi province, China. The specificity of primers of 3 enterotoxins (i.e., cytK, nheA, and hblD) of B. cereus was verified by inclusivity and exclusivity tests using single PCR. Upon optimization of multiplex PCR conditions, it was found that the detection limit of viable cells was 10(2) cfu/mL of B. cereus in pure culture. By enrichment for 3 or 4 h and propidium monoazide pretreatment, a protocol for detection of viable cells as low as 2.2×10(1) cfu/g in spiked food (e.g., milk powder, noodles, and rice) was established and proved valid even under the interference of non-Bacillus cereus at as high as 10(5) cfu/g. Moreover, the protocol based on multiplex PCR for detection was applied for the analysis of distribution of toxin gene of B. cereus, and the results showed a regional feature for toxin gene distribution, indicating that potential toxigenicity of B. cereus should be evaluated further.
Asunto(s)
Bacillus cereus/aislamiento & purificación , Enterotoxinas/aislamiento & purificación , Comida Rápida/microbiología , Microbiología de Alimentos , Fórmulas Infantiles/microbiología , Animales , Bacillus cereus/genética , Enterotoxinas/química , Humanos , Lactante , Leche/química , Leche/microbiología , Reacción en Cadena de la Polimerasa MultiplexRESUMEN
Although the microtubule-stabilizing agent paclitaxel has been widely used for treatment of several cancer types, particularly for the malignancies of epithelia origin, it only shows limited efficacy on hematological malignancies. Emerging roles of O-GlcNAcylation modification of proteins in various cancer types have implicated the key enzymes catalyzing this reversible modification as targets for cancer therapy. Here, we show that the highly selective O-GlcNAcase (OGA) inhibitor thiamet-G significantly sensitized human leukemia cell lines to paclitaxel, with an approximate 10-fold leftward shift of IC50. Knockdown of OGA by siRNAs or inhibition of OGA by thiamet-G did not influence the cell viability. Furthermore, we demonstrated that thiamet-G binds to OGA in competition with 4-methylumbelliferyl N-acetyl-ß-d-glucosaminide dehydrate, an analogue of O-GlcNAc UDP, thereby suppressing the activity of OGA. Importantly, inhibition of OGA by thiamet-G decreased the phosphorylation of microtubule-associated protein Tau and caused alterations of microtubule network in cells. It is noteworthy that paclitaxel combined with thiamet-G resulted in more profound perturbations on microtubule stability than did either one alone, which may implicate the underlying mechanism of thiamet-G-mediated sensitization of leukemia cells to paclitaxel. These findings thus suggest that a regimen of paclitaxel combined with OGA inhibitor might be more effective for the treatment of human leukemia.
Asunto(s)
Leucemia/patología , Microtúbulos/efectos de los fármacos , Paclitaxel/farmacología , Piranos/farmacología , Tiazoles/farmacología , beta-N-Acetilhexosaminidasas/antagonistas & inhibidores , Humanos , Células Jurkat , Leucemia/enzimología , Leucemia/metabolismo , Fosforilación , Proteínas tau/metabolismoRESUMEN
BACKGROUND: Burkitt's lymphoma is an aggressive malignancy with high risk of metastasis to extranodal sites, such as bone marrow and central nervous system. The prognosis of metastatic Burkitt's lymphoma is poor. Here we sought to identify a role of histone deacetylase 6 (HDAC6) in the metastasis of Burkitt's lymphoma cells. METHODS: Burkitt's lymphoma cells were pharmacologically treated with niltubacin, tubacin or sodium butyrate (NaB) or transfected with siRNAs to knock down the expression of HDAC6. Cell migration and invasion ability were measured by transwell assay, and cell cycle progression was analyzed by flow cytometry. Cell adhesion and proliferation was determined by CellTiter-Glo luminescent cell viability assay kit. Cell morphological alteration and microtubule stability were analyzed by immunofluorescence staining. Effect of niltubacin, tubacin and NaB on acetylated tubulin and siRNA efficacy were measured by western blotting. RESULTS: Suppression of histone deacetylase 6 activity significantly compromised the migration and invasion of Burkitt's lymphoma cells, without affecting cell proliferation and cell cycle progression. Mechanistic study revealed that HDAC6 modulated chemokine induced cell shape elongation and cell adhesion probably through its action on microtubule dynamics. CONCLUSIONS: We identified a critical role of HDAC6 in the metastasis of Burkitt's lymphoma cells, suggesting that pharmacological inhibition of HDAC6 could be a promising strategy for the management of metastatic Burkitt's lymphoma.
RESUMEN
BACKGROUND: Although rituximab in the combination of CHOP chemotherapy has been widely used as the standard treatment for several kinds of B-cell non-Hodgkin lymphoma (B-NHL), a great number of B-NHL patients treated with this immunotherapy still develop primary and secondary resistance. Recently Bruton's tyrosine kinase (Btk) inhibitor ibrutinib showed promising therapeutic effect in relapsed/refractory CLL and B-cell NHL, which provided essential alternatives for these patients. METHODS: The proliferation and apoptosis induction of tumor cells were measured by cell viability assay and Annexin-V staining. Western Blotting analysis and real-time PCR were used to detect the expression level of target proteins and chemokines production. RESULTS: We demonstrated that ibrutinib inhibited the proliferation and induced apoptosis of GCB-DLBCL cell lines through suppression of BCR signaling pathway and activation of caspase-3. Furthermore, the chemokines CCL3 and CCL4 production from tumor cells were also found to be attenuated by ibrutinib treatment. But different cell lines exhibited distinct sensitivity after ibrutinib treatment. Interestingly, the decreasing level of p-ERK after ibrutinib treatment, but not the basal expression level of Btk, correlated with different drug sensitivity. CONCLUSIONS: Ibrutinib could be a potentially useful therapy for GCB-DLBCL and the decreasing level of p-ERK could become a useful biomarker to predict related therapeutic response.