Additional file 1 of Risk factors associated with severe adverse events in patients with relapsing polychondritis undergoing flexible bronchoscopy

Supplementary Material

EEG and histological measures from aging mice in the untreated/post-CS treated or prophylactically treated cohort.

<p>Post-CS and prophylactic anticonvulsive treatments were conducted in two separate cohorts of aging mice (n = 24 and 21 respectively). <b>Panel A</b>: Changes in ipsilateral hippocampal EEG signals were analyzed in 8 and 5 animals with CS from these two cohorts respectively. Ipsilateral hippocampal EEG signals were normalized as a % of the baseline. The numbers of animals examined at different post-HI time points are indicated in the parentheses. <b>Panel B</b>: ipsilateral EEG signals were analyzed in untreated animals and prophylactically treated animals (n = 6 each) that did not exhibit CS and ipsilateral infarctions on later histological assessment. Data are similarly presented in Panel A. <b>Panel C</b>: The ratio of the ipsilateral to contralateral hemispheric areas measured in histological brain sections. These measurements were made in 4 untreated/post-CS treated animals and 7 prophylactically treated animals 4–5 weeks post-HI. There were no significant group differences in any EEG or histological measures (p>0.05, t test or Mann-Whitney Rank Sum Test).</p><p>EEG and histological measures from aging mice in the untreated/post-CS treated or prophylactically treated cohort.</p

Abundant Fluoro-Jade positive cells in aging mice with post-HI CS.

<p><b>A-F,</b> representative images were collected from 2 aging mice 24 hours following HI, one with CS (D-F, right), the other without (A-C, left). Brain regions in which Fluoro-Jade positive cells were analyzed are indicated diagrammatically in middle column (by red squares). <b>G</b>, an enlarged image was taken from a selected cortical area (indicated by a yellow square in F). <b>H</b>, regional counts of Fluoro-Jade positive cells in 8 post-HI aging mice with and without CS (mean±SE; n = 4 in each group). *, p<0.05, ipsilateral cell counts in animals with CS vs. contralateral cell counts and cell counts in animals without CS, one way ANOVA.</p

Ipsilateral brain injury or infarctions in aging mice with post-HI CS.

<p><b>A</b>-<b>D</b>, images of cresyl violet-stained brain sections were obtained from 4 aging mice. Left, low power views at three coronal levels. Right, magnified views of the dorsal hippocampal areas (denoted by black dots on low power view). Time of histologic preparation, experimental group, and CS occurrence were indicated for each animal above their respective low power images. <b>E</b>, ratios of ipsilateral relative to contralateral hemispheric areas were obtained at 8 coronal levels and averaged for each animal. Data (%, mean±SE) for individual animals were pooled together according to the indicated experimental groups. *, p<0.05, sham controls vs. other experimental groups, one way ANOVA.</p

CS incidence and lack of CS-concurrent hippocampal-cortical EEG discharges.

<p><b>A</b>, a schematic presentation of experimental manipulations and animal groups. <b>B</b>, CS incidences in aging mice following HI, sham surgery or hypoxia alone. <b>C</b>, representative brain histologic sections obtained from one aging mouse, showing the tracks (black arrows) of implanted EEG electrodes in the ipsilateral hippocampus (right) and contralateral cortex (left). Similar observations were made in other 9 animals to confirm the location of implanted EEG electrodes. <b>D-E</b>, representative EEG traces were collected from 2 aging mice before, during, and shortly after hypoxia. Tethered recordings were made from the ipsilateral hippocampus (ipsil. hipp.) and contralateral parietal cortex (contra. cort.). Original data were treated with band-pass filtered (0.5–500 Hz) for illustration purpose. Red circles or green squares denote movement artifacts in the absence or presence of CS. Arrowed segments are expanded below, showing hippocampal spikes (D, left) and movement artifact-contaminated signals during CS (D, right and E, right). <b>F</b>, signals of ipsilateral cortical EEG (top) and gross motor activity (bottom) collected telemetrically from another aging mouse. Left, evident motor activity signals without corresponding EEG discharges during a CS event. Right, EEG spikes in the absence of motor activity signals (during immobility).</p

Decreases of ipsilateral EEG signals in aging mice with early-onset CS.

<p><b>A</b>-<b>C</b>, representative EEG traces collected from 3 aging mice during baseline monitoring (left), at the end of ambient air exposure or hypoxia (middle), and 24 hours later (right). Tethered recordings were made from the ipsilateral hippocampus and contralateral parietal cortex for each animal. Note the ipsilateral EEG suppression in the animal with post-HI CS (B), recovered ipsilateral EEG in the animal without CS (C) and the lack of EEG suppression in the control animal (A). <b>D</b>, 30-sec EEG segments were collected during baseline monitoring, at the end of either hypoxia or ambient air exposure, then at 1 hour, and 24 hours following either a sham operation or HI. The root mean square (RMS) of the EEG signals was calculated and normalized as a percentage of the baseline RMS. Animals are grouped as sham controls and post-HI with and without CS. Data (mean±SE) for the ipsilateral hippocampus (left) and the contralateral cortex (right) are presented separately. *, p<0.05, sham control vs. others, one way ANOVA.</p

Ipsilateral brain injuries observed histologically from individual aging mice.

<p>Ipsilateral brain injury was recognized through hypochromic staining when examined at 24–48 hours post-HI or cystic infarctions at 4–5 weeks post HI. Column 1: Animal ID of each aging mouse examined. Columns 2–6: injured brain structures indicated by ‘+’. Abbreviations: HIPP—hippocampus; LAT CORT—lateral cortex; STR—striatum; TH—thalamus; MB/BST—midbrain and brainstem areas. Column 7: ipsilateral regions with hypochromic staining where clearly recognized boundaries were present and quantifiable in 6 animals. These regions were measured at 8 coronal levels (bregma 1.9, 1.2, 0.5, -0.2, -1.1, -1.5, -2.4 and -3.2 mm respectively) and normalized as a % of total ipsilateral hemispheric area. The mean±SE from multiple coronal levels were presented for each animal. Column 8: ipsilateral brain injury observed for other animals in which injury margins were difficult to demarcate. For these animals, the coronal levels at which hypochromic staining or infarctions were observed were indicated for each animal instead.</p><p>Ipsilateral brain injuries observed histologically from individual aging mice.</p