Mycobacterium tuberculosis Cell Wall Antigens Induce the Formation of Immune Complexes and the Development of Vasculitis in an Experimental Murine Model.

Tuberculosis (TB) of the central nervous system (CNS) presents high mortality due to brain damage and inflammation events. The formation and deposition of immune complexes (ICs) in the brain microvasculature during Mycobacterium tuberculosis (Mtb) infection are crucial for its pathobiology. The relevance of ICs to Mtb antigens in the pathogenesis of CNS-TB has been poorly explored. Here, we aimed to establish a murine experimental model of ICs-mediated brain vasculitis induced by cell wall antigens of Mtb. We administered a cell wall extract of the prototype pathogenic Mtb strain H37Rv to male BALB/c mice by subcutaneous and intravenous routes. Serum concentration and deposition of ICs onto blood vessels were determined by polyethylene glycol precipitation, ELISA, and immunofluorescence. Histopathological changes in the brain, lung, spleen, liver, and kidney were evaluated by hematoxylin and eosin staining. Our results evidenced that vasculitis developed in the studied tissues. High serum levels of ICs and vascular deposition were evident in the brain, lung, and kidneys early after the last cell wall antigen administration. Cell wall Mtb antigens induce strong type III hypersensitivity reactions and the development of systemic vasculitis with brain vascular changes and meningitis, supporting a role for ICs in the pathogenesis of TB.

Investigation of EBT3 radiochromic film's response to humidity.

PURPOSE: The aim of this work is to investigate the effects of immersing EBT3 radiochromic film in water and to evaluate its contribution to the total uncertainty in dose determination. MATERIALS AND METHODS: We used 3 cm × 3 cm EBT3 radiochromic films irradiated in the range of 0-70 Gy to study the impact of water immersion on the change in net optical density. These films were placed in a water container for a period of 24 h. The net optical density was measured before (0 h) and after of the immersion in water (1, 3, 6, 12, 18, and 24 h). The absorbance spectrum of the EBT3 radiochromic film was measured at 0 h and 24 h after immersion in water. The uncertainty in dose determination due to the effects of keeping the EBT3 radiochromic film submerged in water at 0, 1, and 24 h were recorded in the red, green, and blue channels. RESULTS: We observed an increase in the net optical density as an effect on the film due to its immersion in water. The penetration of the water at the edges of the radiochromic film was observed to be a function of time during which the film remained in the water. On the other hand, the penetration of water at the edges of the film was found to be independent of irradiation dose. CONCLUSIONS: EBT3 radiochromic film is found more resistant to water penetration through the edges than its predecessors. However, there is evidence that suggest that liquid water damage the Nylon cover layer of the film by changing its optical properties. Therefore, it is recommended to build a new calibration curve for radiochromic films for a specific situation involving dose measurements in liquid water.

Spectral analysis of the EBT3 radiochromic films for clinical photon and electron beams.

PURPOSE: The purpose of this study was to investigate the spectral response of the EBT3 radiochromic films to different beam qualities for radiation therapy dosimetry. Dose, dose rate, and interbatch dependencies on the spectral response of the films were investigated as well. METHODS: Pieces of EBT3 films placed between layers of solid water phantoms were irradiated with 6 and 15 MV photon beams, 6 and 10 MV-flattening filter free (FFF) photon beams, and 6 and 20 MeV electron beams at dose levels between 0.4 and 50 Gy. Net absorbance was measured as a function of wavelength from the spectra acquired in the wavelength range of 400-800 nm using a fiber-coupled spectrometer and broadband light source. RESULTS: No significant change was observed in the absorption spectra of the EBT3 film from the same batch irradiated with the same amount of dose using different beam qualities. Also, no spectral change with dose rate was observed. The measured net absorbance per Gy was independent of beam quality in the 1-50 Gy dose range. Slight differences in the spectral shape and absorption band positions were observed in film samples from different batches. The net absorbance spectra showed two absorption bands centered around 634-636 nm (primary) and 583-585 nm (secondary). However, depending on the film batch, for doses above a certain level the primary absorption band appears to "split" into two bands centered around ~624-628 and ~641-645 nm. CONCLUSIONS: The spectral shape of the EBT3 radiochromic films irradiated with photons (including FFF) and electron beams is beam quality and dose rate independent; however, it varies with dose level, batch, and spectroscopy system used.

On the spectral characterization of radiochromic films irradiated with clinical proton beams.

Radiochromic films have been widely studied for clinical dosimetry in conventional external beam radiation therapy. With an increase in practice of proton therapy, such films are being conveniently used; however, their spectroscopic characterization for this modality is lacking. This work investigated the response of the EBT3 radiochromic films irradiated in a Mevion S250™ clinical proton beam. Dose, dose rate, inter-batch, sensitivity, and linear energy transfer (LET) dependencies of the films were studied. Pieces of the radiochromic films from different batches were irradiated using a spread-out Bragg peak (SOBP) proton beam at dose levels between 0.1-15 Gy. Absorption spectra were measured in the wavelength range of 400-800 nm with 2.5 nm resolution. For comparison, the optical density of the films was measured using a flatbed scanner. The net absorbance spectra showed two characteristic absorption bands centered at 636 nm and 585 nm. However, a saturation effect, manifested as broadening/splitting appearance, was observed in the 636 nm band for doses beyond a certain batch-dependent level ~4-10 Gy, in the three different film batches studied. The differences in the spectral shape led to dose-response curves with variable sensitivity. In general a high spectral sensitivity was observed in 0.1-6 Gy range for the three film batches. For a given dose, no significant change in the spectra was observed with change in the dose rate. No significant dependency on the LET was observed for the EBT3 films irradiated with proton beams with dose-averaged LETs ranging from 1.14-6.50 keV µm-1 studied in this work. However, at a given dose, ~5% lower spectral response was observed in the films irradiated with protons compared to their counterparts irradiated with photon beams.