Supplementary Material, AJMQ_TQA_SUPPLEMENTARY_MATERIAL – The Teachers of Quality Academy: Evaluation of the Effectiveness and Impact of a Health Systems Science Training Program

<p>Supplementary Material, AJMQ_TQA_SUPPLEMENTARY_MATERIAL for The Teachers of Quality Academy: Evaluation of the Effectiveness and Impact of a Health Systems Science Training Program by Danielle S. Walsh, Suzanne Lazorick, Luan Lawson, Donna Lake, Herbert G. Garrison, Jason Higginson, Paul Vos, and Elizabeth Baxley in American Journal of Medical Quality</p

<i>Bifidobacterium breve</i> Attenuates Murine Dextran Sodium Sulfate-Induced Colitis and Increases Regulatory T Cell Responses

<div><p>While some probiotics have shown beneficial effects on preventing or treating colitis development, others have shown no effects. In this study, we have assessed the immunomodulating effects of two probiotic strains, <i>Lactobacillus rhamnosus (L. rhamnosus)</i> and <i>Bifidobacterium breve (B. breve)</i> on T cell polarization <i>in vitro</i>, using human peripheral blood mononuclear cells (PBMC), and <i>in vivo</i>, using murine dextran sodium sulfate (DSS) colitis model. With respect to the latter, the mRNA expression of T cell subset-associated transcription factors and cytokines in the colon was measured and the T helper type (Th) 17 and regulatory T cell (Treg) subsets were determined in the Peyer's patches. Both <i>L. rhamnosus</i> and <i>B. breve</i> incubations <i>in vitro</i> reduced Th17 and increased Th2 cell subsets in human PBMCs. In addition, <i>B. breve</i> incubation was also able to reduce Th1 and increase Treg cell subsets in contrast to <i>L. rhamnosus</i>. <i>In vivo</i> intervention with <i>B. breve</i>, but not <i>L. rhamnosus</i>, significantly attenuated the severity of DSS-induced colitis. In DSS-treated C57BL/6 mice, intervention with <i>B. breve</i> increased the expression of mRNA encoding for Th2- and Treg-associated cytokines in the distal colon. In addition, intervention with <i>B. breve</i> led to increases of Treg and decreases of Th17 cell subsets in Peyer's patches of DSS-treated mice. <i>B. breve</i> modulates T cell polarization towards Th2 and Treg cell-associated responses <i>in vitro</i> and <i>in vivo</i>. <i>In vivo B. breve</i> intervention ameliorates DSS-induced colitis symptoms and this protective effect may mediated by its effects on the T-cell composition.</p></div

<i>B. breve</i> intervention changes mRNA expression of Th2-, Th17- and Treg- associated cytokines in the colon.

<p>The mRNA expression of Th1- (<i>Ifnγ</i> and <i>Il12</i>), Th2- (<i>Il4, Il5</i> and <i>I13</i>), Th17- (<i>Il23</i> and <i>Il17</i>) and Treg- (<i>Tgfβ</i> and <i>Il10</i>) associated cytokines was quantified in the distal colons of both A) healthy and B) DSS-treated mice with or without <i>B. breve</i> intervention. Results are expressed as mean + SEM, n = 5 mice per group, pooled from two independent experiments. * p<0.05; ** p<0.01; *** p<0.001.</p

<i>B. breve</i>, but not <i>L. rhamnosus</i>, ameliorates DSS-induced colitis.

<p>C57BL/6 mice with or without probiotics treatment received either normal drinking water or drinking water with DSS for 5 days. A) The fecal condition was calculated on day 0, day 3 and day 5 after DSS treatment. On day 6, the mice were sacrificed and B) the colon length of each mouse was measured. Results are expressed as mean ± SEM, n = 6 mice per group, pooled from two independent experiments. Colons were collected and examined for histological score as described in materials and methods. C) The histological scoring graph and D) representative H&E staining photos are shown. Results are expressed as mean + SEM, n = 3 mice per group, pooled from two independent experiments. E) The presence of Ly-6B+ cells was visualized in the proximal (p) and distal (d) colons using immunohistochemistry. The pictures are representative of 3 separate mice per group obtained from two experiments. F) The concentration of MPO was measured in colon homogenates of each group. Results are expressed as mean + SEM, n = 4 mice per group, pooled from two independent experiments. * p<0.05; ** p<0.01.</p

<i>L. rhamnosus</i> and <i>B. breve</i> alter T cell differentiation in human PBMCs.

<p>PBMCs were stimulated with anti-CD3 alone (white bars), with a combination of anti-CD3 and <i>L. rhamnosus</i> (grey bar) or a combination of anti-CD3 and <i>B. breve</i> (black bar) for 48 hours or 7 days. A–D) The percentages of Th2 (GATA3+Tbet-), Th17 (RORγ+FOXP3-), Treg (RORγ-FOXP3+) or Th1 (GATA-Tbet+) cells within the activated T cells (CD4+CD69+) in the PBMCs were determined after 48 hours of incubation. Percentages within activated CD4+CD69+ T cell population are shown. E–H) The percentages of cytokines (IL10, IL17, IL4 or IFNγ) producing CD4+ T cells in the PBMCs were determined after 7 days of incubation. Percentages within CD4+ T cell population are shown. The Results are expressed as mean + SEM, n = 3, * p<0.05.</p

qPCR primer sequences.

<p>qPCR primer sequences.</p

Treatment toxicity reported in studies using IMRT and chemotherapy for the treatment of locally advanced rectal cancer.

<p>Treatment toxicity reported in studies using IMRT and chemotherapy for the treatment of locally advanced rectal cancer.</p

Acute and late toxicities following intensity-modulated and image-guided radiotherapy for 27 patients with locally advanced laryngeal cancer.

<p>Acute and late toxicities following intensity-modulated and image-guided radiotherapy for 27 patients with locally advanced laryngeal cancer.</p

Dose distribution to target volume and to critical organs at risk for complications following image-guided radiotherapy for head and neck cancer.

<p>PTV1: target volume receiving 66 to 70 Gy; PTV2: target volume receiving 59.6 to 63 Gy; PTV3: target volume receiving 54 to 56 Gy; Gy: gray.</p

Aspiration rate reported in the literature following radiotherapy for non-laryngeal and non-hypopharyngeal head and neck cancer.

<p>C: conventional with two lateral and a supraclavicular field; NS: not specified; IMRT: intensity-modulated radiotherapy; WF: whole-field; SF: split-field; IGRT: image-guided radiotherapy.</p