Supplementary Material for: Low Birthweight and Premature Birth Are Risk Factors for Podocytopenia and Focal Segmental Glomerulosclerosis

<b><i>Background:</i></b> Recent reports suggest that low birthweight (LBW) is a risk factor for kidney diseases, including focal segmental glomerulosclerosis (FSGS), although the underlying pathological mechanism remains unknown. Podocyte loss triggers glomerulosclerosis; however, whether FSGS in LBW children is associated with podocytopenia is unclear. <b><i>Methods:</i></b> We reviewed the birthweights and gestational age of all patients who underwent renal biopsies from 1995 to 2011 at our Institute. Sixteen patients had FSGS, of which 6 (37.5%) had LBW; this LBW rate was significantly higher than the overall LBW rate in Japan (9.7%). The incidence of LBW was also high in patients with minimal change nephrotic syndrome (MCNS; 12.5%). The glomerular cell numbers in biopsy sections were calculated using computer image analysis and compared with FSGS of normal birthweight (NBW-FSGS). Biopsy specimens from age-matched patients with MCNS were also compared. Wilms' tumor-1 (WT1) immunohistochemistry was performed to enumerate the podocytes. <b><i>Results:</i></b> All patients in the LBW-FSGS group were also preterm, with an average gestational age of 25.8 weeks. The number of podocytes per glomerulus in the LBW-FSGS patients was 34 and 24% lower as compared to that in the MCNS patients (p < 0.01) and the NBW-FSGS patients (p < 0.05), respectively. Similar results were observed for the WT1-positive glomerular cell number. <b><i>Conclusion:</i></b> LBW and premature birth were associated with FSGS development. The possibility that LBW and premature birth may be predisposing factors for severe podocytopenia in children with FSGS warrants further investigation

Supplementary Material for: Guggulsterone, a Plant-Derived Inhibitor of NF-TB, Suppresses CDX2 and COX-2 Expression and Reduces the Viability of Esophageal Adenocarcinoma Cells

<b><i>Background/Aims:</i></b> Induction by bile acid of caudal type homeobox 2 (CDX2) and cyclooxygenase-2 (COX-2) expression via nuclear factor-TB (NF-TB) activation is a critical event in the development of Barrett's esophagus (BE) and esophageal adenocarcinoma (EAC). Guggulsterone (GS) is a plant sterol that inhibits NF-TB activity. Here, we evaluated whether GS has either or both chemopreventive or therapeutic effects against EAC. <b><i>Methods:</i></b> Two EAC cells lines were treated with deoxycholic acid (DCA) in the presence of GS or vehicle. The levels of transcription and translation of <i>ITBE</i>, <i>CDX2</i>, and <i>COX-2</i> were determined. Prostaglandin E₂ (PGE₂) levels, cell viability, and cell cycle distribution were assessed as well. <b><i>Results:</i></b> GS inhibited DCA-induced ITBE phosphorylation. GS and the NF-TB inhibitor BAY11-7085 suppressed DCA-induced <i>CDX2</i> and <i>COX-2</i> expression in EAC cells. GS also suppressed basal transcription levels of <i>CDX2</i> and <i>COX-2</i> and reduced constitutive synthesis of COX-2 and PGE₂. Further, GS reduced the viability of EAC cells, increased their numbers in the apoptotic sub-G1 fraction. <b><i>Conclusion:</i></b> GS suppressed DCA-induced and NF-TB-dependent activation of <i>CDX2</i> and <i>COX-2</i> expression. Further, GS also reduced the viability of EAC cells. GS may serve as candidate for preventing and treating EAC and BE. i 2014 S. Karger AG, Base

Supplementary Material for: Narrow Band Imaging with Magnifying Endoscopy for Peyer's Patches in Patients with Inflammatory Bowel Disease

<b><i>Background/Aims:</i></b> Peyer's patches (PPs) play a major role in mucosal immunity, but little is known about their alterations in patients with inflammatory bowel disease (IBD). We aimed to evaluate endoscopic changes of PPs in IBD patients using narrow band imaging with magnifying endoscopy (NBI-ME). <b><i>Methods:</i></b> Images of PPs using NBI-ME by ileocolonoscopy were consecutively collected. Existence of branch-like structures and the vessel occupancy in the dome lesions of PPs were analyzed. Appearance of the surrounding villi of the domes in PPs was evaluated using a ‘villi index' consisting of irregular formation, hyperemia, and altered vascular network pattern. Vascularity of PPs was immunohistologically analyzed by anti-CD34 antibody. <b><i>Results:</i></b> 17 patients with Crohn's disease (CD), 43 with ulcerative colitis (UC), and 23 healthy control subjects (HC) were analyzed. Both CD and UC patients had a high prevalence of having branch-like structures and significantly higher vascularity in the dome lesions than HC. The villi indices and vascular widths in the villi were significantly larger in CD and UC patients than in HC. <b><i>Conclusions:</i></b> Precise examination with NBI-ME characterized alteration of vascular structure in the dome and surrounding villi lesions of PPs not only in CD but also in UC patients

Supplementary Material for: MicroRNA Expression and Functional Profiles of Osteosarcoma

<b><i>Objective:</i></b> Osteosarcoma (OS) is the most frequent primary malignant bone tumor in children and young adults. Although the introduction of combined neoadjuvant chemotherapy has significantly prolonged survival, the outcome for OS patients showing a poor response to chemotherapy is still unfavorable. In order to develop new therapeutic approaches, elucidation of the entire molecular pathway regulating OS cell proliferation would be desirable. <b><i>Methods:</i></b> MicroRNA (miRNA) are highly conserved noncoding RNA that play important roles in the development and progression of various other cancers. Using miRNA microarrays capable of detecting a known number of 933 miRNA, 108 miRNA were found to be commonly expressed in 24 samples of OS tissue and subjected to a cell proliferation assay. <b><i>Results:</i></b> We found that inhibition of 5 let-7 family miRNA (hsa-let-7a, b, f, g and i) significantly suppressed the proliferation of OS cells. Using a quantitative shotgun proteomics approach, we also found that the let-7 family miRNA regulated the expression of vimentin and serpin H1 proteins. <b><i>Conclusions:</i></b> Our present results indicate the involvement of let-7 family miRNA in regulation of the cell proliferation as well as epithelial-mesenchymal transition of OS. Thus, let-7 family miRNA may potentially provide novel targets for the development of therapeutic strategies against OS