Supplementary Material for: Dynamic Prognostic Score to Predict Survival in East Asian Elderly Populations with Dysphagia

Introduction: Dysphagia is a common disorder in older adults leading to severe complications, including aspiration pneumonia, dehydration, malnutrition, weight loss, and even death. However, no prognostic model has been developed to predict the prognosis of older adults with dysphagia. Methods: Data from patients with dysphagia at a single center were retrospectively reviewed between 2014 and 2017. All data were obtained from the Dryad Digital Repository. The least absolute shrinkage and selection operator regression model was used to select potentially relevant features. Multiple Cox proportional hazard analysis was used to develop a model based on the training set. Results: The nomogram comprised age, sex, percutaneous endoscopic gastrostomy, chronic heart failure, total lymphocyte count, daily calorie intake, and severe pneumonia, which provided favorable calibration and discrimination in the training dataset, with AUCs for the 1-year, 2-year, and 3-year survival predictions of 0.833, 0.871, and 0.886, respectively. Furthermore, it showed acceptable discrimination in the validation cohort, with AUCs for the 1-year, 2-year, and 3-year survival predictions of 0.884, 0.834, and 0.782, respectively. Moreover, the decision curve analysis results revealed that the nomogram was clinically beneficial. Conclusion: A nomogram, combining seven demographic and clinical factors, provided an excellent preoperative prediction of survival probability in older individuals with dysphagia. This predictive model can be used as a reference to assist clinicians in making clinical decisions

Supplementary Material for: Colonization of <b><i>Caenorhabditis elegans</i></b> by <b><i>Bacillus nematocida</i></b> B16, a Bacterial Opportunistic Pathogen

<b><i>Background:</i></b> Soil-dwelling <i>Bacillus nematocida</i> B16 can kill <i>Caenorhabditis elegans</i> via a Trojan horse-like mechanism. However, colonization is a key problem that must be solved during the infection process. <b><i>Aims:</i></b> To study the molecular mechanism involved in the colonization of <i>B. nematocida</i> B16 against the host <i>C. elegans</i>. <b><i>Methods:</i></b> GFP-expressing strain B16g was constructed and its nematocidal activity was assayed. ‘Feeding transfer’ experiments were carried out separately using of B16 and B16g strains to explore the colonization mode of the bacteria. Fluorescence microscopy was used to observe the interactions between fluorescent signal and the quantity of bacteria in the intestine. A mariner-based transposon called TnYLB-1 was also applied in the random mutagenesis of B16 to screen the mutants with impaired colonization of nematode worms and identify potential localization-related genes. <b><i>Results and Conclusion:</i></b> A small inoculum of the bacteria resulted in its proliferation in the <i>C. elegans</i> intestine. The fluorescence signal was enhanced with increasing bacterial density in the intestine. Several candidate genes with possibly important roles in colonization were found. These results provide a solid foundation for further elucidation of the infection process at the molecular level and enrichment of our knowledge of bacterial pathogenesis

Erratum: Rare RET Variant p.D707E in a Chinese Pedigree with Hereditary Medullary Thyroid Carcinoma

<b><i>Background:</i></b> Hereditary medullary thyroid carcinoma (HMTC) is thought to be associated with germline mutations of the RET proto-oncogene. <b><i>Methods:</i></b> We detected RET proto-oncogene germline mutations from a pedigree with HMTC in the east of China and investigated the characteristics of these mutations in this pedigree and their correlation with HMTC by direct sequencing of all 21 exons in the RET gene of all 46 subjects. <b><i>Results and Conclusion:</i></b> (1) Thirteen types of RET gene variants were detected in this pedigree. Of these, p.F285S in exon 4, c.854_855CA in exon 4, and p.D707E in exon 11 are reported for the first time in our study. (2) Both linkage disequilibrium analysis and logistic regression analysis showed a significant correlation between the p.D707E variant and HMTC (LOD = 3.69, OR = 4.413, p = 0.000167), indicating that this variant is a risk factor for medullary thyroid carcinoma (MTC). (3) The single-nucleotide polymorphisms (SNP) G691S in exon 11 (rs1799939), S904S in exon 15 (rs1800863), and rs2075912 and rs2565200 in the 3′-untranslated region of the RET proto-oncogene are in complete linkage disequilibrium (D' = 1, r² = 1); no correlation of these SNP and MTC was observed in this pedigree. (4) No hot-spot mutation of the RET proto-oncogene was detected in this pedigree. We drew the conclusion that the heterozygous nonsynonymous variant p.D707E in the RET proto-oncogene is rare, but it is a risk factor for hereditary MTC

Supplementary Material for: Treatment of Secondary Hyperparathyroidism: Results of a Phase 2 Trial Evaluating an Intravenous Peptide Agonist of the Calcium-Sensing Receptor

<p><b><i>Background/Aims:</i></b> This study evaluated the efficacy and safety of AMG416 (etelcalcitide), a novel peptide agonist of the calcium (Ca)-sensing receptor given intravenously (IV) after each hemodialysis session for the treatment of secondary hyperparathyroidism (SHPT). <b><i>Methods:</i></b> Adult subjects with SHPT on hemodialysis enrolled in a 12-week, dose titration (parent) study followed by an open-label extension phase. AMG416 was administered IV, thrice weekly starting at 5 mg/session and titrated based on the subject's parathyroid hormone (PTH) and albumin-corrected Ca (cCa) to target a PTH of 150-300 pg/ml. Efficacy (percent PTH change from baseline to the efficacy analysis period during the parent study) and safety (open-label extension phase) endpoints were evaluated. <b><i>Results:</i></b> Baseline (n = 37) mean (standard error [SE]) PTH was 853 (106 pg/ml). The mean (95% CI) percent change from baseline to the efficacy analysis period in PTH concentration was -53.6% (-60.8, -46.4). The proportion of subjects with ≥30% reduction in PTH from baseline to the efficacy assessment period (EAP) was 89% (32/36; 95% CI 73.9, 96.9). Results by the baseline PTH subgroup (≤700 vs. >700 pg/ml) were comparable for both analyses. The proportion of subjects achieving a PTH ≤300 pg/ml was 56% (n = 20/36) at the efficacy assessment period. The mean (SE) percent changes from baseline to EAP were observed for cCa -15% (1.0%) and phosphorus -10% (3.3%). Adverse events were mild to moderate in severity. The PTH reductions achieved in the parent study were maintained in the open-label extension phase. <b><i>Conclusion:</i></b> AMG416 was well tolerated and appears to be an effective agent for the treatment of SHPT in patients on hemodialysis.</p

Supplementary Material for: CyclinG1 Amplification Enhances Aurora Kinase Inhibitor-Induced Polyploid Resistance and Inhibition of Bcl-2 Pathway Reverses the Resistance

<i>Background/Aims:</i> CyclinG1 (CycG1) is frequently overexpressed in solid tumors and overexpression of CycG1 promotes cell survival upon paclitaxel exposure by inducing polyploidy. Whether and how CycG1 regulates polyploidization caused by small molecular targeted inhibitors remains unclear. <i>Methods:</i> Immunohistochemistry and immunoblotting were utilized to examine protein expression. Cell proliferation was measured by ATPlite assay, and cell cycle distribution and apoptosis were measured by flow cytometry and/or DNA fragmentation assays. <i>Results:</i> Overexpression of CycG1 in breast cancer cells caused apoptosis-resistant polyploidy upon treatment with Aurora kinase inhibitor, ZM447439 (ZM). Addition of ABT-263, a small-molecule BH3 mimetic, to ZM, produced a synergistic loss of cell viability with greater sustained tumor growth inhibition in breast cancer cell lines. Decrease of Mcl-1 and increase of NOXA caused by ZM treatment, were responsible for the synergy. Furthermore, CycG1 was highly expressed in Triple-Negative-Breast-Cancer patients treated with paclitaxel and was paralleled by decreased cell survival. <i>Conclusion:</i> CycG1 is a crucial factor in ZM-induced polyploidy resistance, and ABT-263/ZM combination hold therapeutic utility in the CycG1-amplified subset of breast cancer and CycG1, thus, is a promising target in breast cancer