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Additional file 2: Figure S2. The three animal experimental protocols used in this study. The high-concentration intermittent (repeated) inhalation exposure study (A), the high-concentration single inhalation study (B), and the repeated intratracheal instillation study (C).

Additional file 1: Fig. S1. The whole body inhalation exposure system using in this study. The whole body inhalation exposure system (A), the averaged CWAAP-A concentration in the chamber per each exposure day (B), cumulative frequency distribution graphs with logarithmic probability (C) and representative scanning electron microscope (SEM) images of the CWAAP-A particles in the chambers (D). Scale bar: 20 μm (panel D).

Additional file 5: Fig. S5. The effect of pretreatment of the CWAAP-A-exposed rats with clodronate liposomes (to deplete macrophages) or polymorphonuclear leukocytes (PMN)-neutralizing antibodies (to deplete neutrophils) on BALF markers and cell populations. The lactate dehydrogenase (LDH) activity (A) and surfactant protein-D (SP-D) in the BALF B, and the number of lymphocytes (C), eosinophils (D) and basophils € in the plasma are shown. Tukey’s multiple comparison test: *p < 0.05, **p < 0.01, and ***p < 0.001, pairs indicated.

Additional file 4: Fig. S4. Co-localization of vascular endothelial growth factor receptor 3 (VEGFR3) with lymphatic vessels in rat lungs. Lung sections from Prox1-EGFP transgenic rats were stained with three commercially available lymphatic markers, VEGFR3, lymphatic vessel endothelial hyaluronan receptor 1 (Lyve-1), and podoplanin (RT1-40), with co-staining of GFP and 4',6-diamidino-2-phenylindole (DAPI; a nucleus marker). Only VEGFR3 co-localized almost completely with GFP in the rat lungs, including in the peripheral regions. Br Bronchus and Vn Venule.

Additional file 6: Fig. S6. Design of animal experimental protocols in this research. 13-week inhalation exposure study (A), and macrophages or neutrophils depletion study (B).

Additional file 2: Fig. S2. Representative macroscopic photographs of cross-sections of rat lungs. The left and right sides represent males and females, respectively. conc concentration, expo exposure, rec recovery.

Additional file 3: Fig. S3. Visualization of normal lymphatic vessels in the rat lung using Prospero homeobox protein-1 (Prox1)-enhanced green fluorescent protein (EGFP) transgenic rats [13]. (A) Lung sections from Prox1-EGFP transgenic rats were double-stained with GFP and α-smooth muscle actin (αSMA) and counterstained with hematoxylin. Lymphatic vessels (red arrows) run around the veins and arteries. Blue arrows represent neuroendocrine cells. (B) A graphical image of lymphatic vessels running (green lines) through the lungs.

Figure S1. Sequence data of internal transcribed spacer (ITS) region. Figure S2. Sequence data of domain 1 and 2 region. Figure S3. Sequence data of glyceraldehyde-3-phosphate dehydrogenase (G3PDH) gene. Figure S4. Sequence data of heat-shock protein 60 (HSP60) gene. Figure S5. Sequence data of DNA-dependent RNA polymerase subunit II (RPB2) gene. (PDF 383 kb)

Purpose: Although TI-201 SPECT has been used to evaluate the malignant grade of cerebral gliomas, the gold standard continues to be histopathologic examination. The authors assessed and compared the prognostic abilities of the two studies using survival analysis. Materials and Methods: Twenty-nine patients underwent 34 sessions of TI-201 SPECT plus surgery for primary or recurrent cerebral gliomas 12 to 78 months before this analysis. Using conventional survival analyses, such as the log-rank test, Cox regression, and the Akaike cross-tab method, the authors evaluated the prognostic significance of 10 variables: histopathologic grade, TI-201 SPECT, Tc-99m HMPAO SPECT, tumor cell viability, radionecrosis, neurologic defects, clinical improvement, surgery, chemotherapy, and external beam radiotherapy. Results: TI-201 SPECT was most strongly related to prognosis, followed by histopathologic grade. The other variables had little prognostic value. The Cox stepwise selection procedure indicated that TI-201 SPECT was the only independent predictor of outcome, whereas histopathologic analysis was eliminated from the prognostic model. However, the Kaplan-Meier survival curve and the Akaike method indicated that histopathologically low-grade tumors were more closely associated with longer-term survival than were TI-201 low uptake tumors. Conclusions: TI-201 SPECT is not only closely correlated with the histopathologic grade of tumor but is a significantly better predictor of outcome than histopathologic grade. However, histopathologic examination may provide additional information on longer-term survival. TI-201 SPECT is a valuable procedure, especially in patients in whom a histologic diagnosis of possible glioma cannot be made.