RESUMO
We developed a diagnostic tool to differentiate between endophthalmitis and uveitis using Raman spectroscopy. Twenty-two New Zealand rabbits with endophthalmitis induced by Staphylococcus aureus (10 animals), noninfectious uveitis induced by lipopolysaccharide from Escherichia coli (10 animals) and controls (two animals) were analyzed. Twenty-four hours after the eyes were inoculated, iris tissue was dissected and subjected to dispersive Raman spectroscopy using an excitation source at 830 nm and a spectrograph/CCD camera to detect a Raman signal with an integration time of 50 s. With the collected spectra of endophthalmitis and uveitis, we developed a routine to classify spectra in each specimen using principal components analysis, using a leave-one-out cross-validation procedure. The mean Raman spectra of tissues with uveitis and endophthalmitis showed several bands in the region of 800-1800 cm(-1), which have been attributed to nucleic acids, amino acids, proteins, and lipids. The bands at 1004, 1339, and 1555 cm(-1) differed significantly (t-test, p<0.05) between diseases. The principal components PC3 and PC4 differed significantly (ANOVA, p<0.05) for the two tissue types, indicating that these PCs can be used to discriminate between the two diseases using Mahalanobis distance as a discriminator. This technique is useful for differentiating the spectral bands of uveitis and endophthalmitis, and the diagnostic model showed sensitivity of 89%, specificity of 100%, and accuracy of 92% using the leave-one-out cross-validation procedure. These results may be clinically relevant for differentiating endophthalmitis from uveitis, and this approach may become a noninvasive method to optimize the diagnosis of inflammatory and infectious vitreoretinal diseases.
Assuntos
Endoftalmite/diagnóstico , Infecções por Escherichia coli/diagnóstico , Infecções Oculares Bacterianas/diagnóstico , Doenças da Íris/diagnóstico , Análise Espectral Raman , Infecções Estafilocócicas/diagnóstico , Uveíte/diagnóstico , Animais , Diagnóstico Diferencial , Endoftalmite/microbiologia , Infecções por Escherichia coli/microbiologia , Infecções Oculares Bacterianas/microbiologia , Doenças da Íris/microbiologia , Coelhos , Sensibilidade e Especificidade , Infecções Estafilocócicas/microbiologia , Uveíte/microbiologiaRESUMO
The microenvironment formed by lauroyl and stearoyl derivatives of chitosan in solution has been studied using two fluorescent probes, pyrene and nabumetone. Existence or not of microdomains formed by polymolecular associations, the inherent hydrophobicity of them in aqueous solution, and the influence of degree of substitution (DS) of derivatives were investigated by emission properties of pyrene and strengthened by the photophysical behavior of nabumetone. Additionally, the ratio between the fluorescence intensities of first (approximately 372 nm) to the third (approximately 384 nm) bands of the emission spectrum of pyrene was used to determine the critical aggregation concentration (CAC). In a previous work, it was already reported the characterization of chitosan derivatives by three spectroscopic techniques ((13)C-NMR, (1)H-NMR and infrared), as well as data on the solubility and swelling-index of them. In addition of that, the new results show that the investigated lauroyl and stearoyl derivatives of chitosan are expected to be potential models for applications in the medical field.