Additional file 1: of Bioassay-guided isolation of active anti-adipogenic compound from royal jelly and the study of possible mechanisms

Data on identification and purity assessment of isolated 10-Hydroxy-2-decenoic acid (10-HDA). Figure S1. 1H NMR Spectrum (A) and 13C NMR (500 MHz) spectra (B) of the compound isolated from ethyl acetate fraction of RJ in DMSO-d6; Figure S2. TLC analysis of Royal jelly (RJ), RJ-EA fraction (EA), isolated 10-HDA compound (Com) and standard 10-HDA (Std); Figure S3. Mass spectrometry - structure of the 10-Hydroxy-2-decenoic acid (10-HDA). (DOCX 156 kb

DataSheet1_18KHT01, a Potent Anti-Obesity Polyherbal Formulation.docx

Obesity is a life-threatening metabolic disorder necessitating urgent development of safe and effective therapy. Currently, limited such therapeutic measures are available for obesity. The present study was designed to develop a novel, safe and effective herbal therapy for the management of obesity. A polyherbal formulation (18KHT01) was developed by homogeneously mixing a specific proportion of crude Quercus acutissima (acorn jelly powder), Camellia sinensis (dry leaf buds), and Geranium thunbergii (dry aerial part) along with Citrus limon (fruit juice). Synergistic antioxidant, antiadipogenic, and anti-obesity activities were evaluated by in vitro as well as in vivo studies. In vitro experiments revealed strong synergistic antioxidant and anti-adipogenic activities of 18KHT01. Molecular assessment of 18KHT01 showed significant down-regulation of vital adipogenic factors such as PPARγ, C/EBPα, aP2, SREBP-1c, FAS, and LPL. Based on the results of the preliminary toxicity study, 75 and 150 mg/kg, twice daily doses of 18KHT01 were administered to evaluate anti-obesity activity in diet-induced obese (DIO) C57BL/6J mice model. The major obesity-related parameters such as body weight, weight gain, food efficiency ratio, as well as serum lipid profile were significantly reduced by 18KHT01 with potential synergism. Also, the high-fat diet-induced insulin resistance was suggestively alleviated by the formulation, and thus ameliorated fasting blood glucose. Histological evaluation of liver and white adipose tissue revealed that the significant reduction of fat depositions and thus reduction of these tissue weights. Synergy evaluation experiments exhibited that the 18KHT01 offered strong synergism by improving efficacy and reducing the toxicity of its ingredients. Overall results evidenced the 18KHT01 as a safe and potent anti-obesity herbal therapy.</p

Mucosal-Associated Invariant T Cell Deficiency in Chronic Obstructive Pulmonary Disease

<p>Mucosal-associated invariant T (MAIT) cells have been reported to play an important role in mucosal immunity. However, little is known about the roles of MAIT cells in chronic obstructive pulmonary disease (COPD). The aims of this study were to examine the levels of circulating MAIT cells and their subsets in COPD patients and to investigate the potential relationship between clinical parameters and MAIT cell levels. Forty-five COPD patients and 57 healthy control subjects were enrolled in the study. Circulating MAIT cells and their subset levels in the peripheral blood were measured by flow cytometry. Disease grades were classified according to the GOLD criteria for the assessment of severity of COPD. Circulating MAIT cell levels were found to be significantly reduced in COPD patients. In particular, this MAIT cell deficiency was more prominent in CD8+ and double-negative T cell subsets. Interestingly, elevated serum C-reactive protein level and reduced FEV₁/FVC ratio were associated with MAIT cell deficiency in COPD patients. Furthermore, the circulating MAIT levels were found to be significantly lower in patients with moderate to severe COPD than in patients with mild COPD. Our data shows that MAIT cells are numerically deficient in the peripheral blood of patients with COPD. In addition, this MAIT cell deficiency was found to reflect inflammatory activity and disease severity. These findings provide important information for monitoring the changes in MAIT cell levels and for predicting the prognosis during the disease course.</p

Activation, Impaired Tumor Necrosis Factor-α Production, and Deficiency of Circulating Mucosal-Associated Invariant T Cells in Patients with Scrub Typhus

<div><p>Background</p><p>Mucosal-associated invariant T (MAIT) cells contribute to protection against certain microorganism infections. However, little is known about the role of MAIT cells in <i>Orientia tsutsugamushi</i> infection. Hence, the aims of this study were to examine the level and function of MAIT cells in patients with scrub typhus and to evaluate the clinical relevance of MAIT cell levels.</p><p>Methodology/Principal Findings</p><p>Thirty-eight patients with scrub typhus and 53 health control subjects were enrolled in the study. The patients were further divided into subgroups according to disease severity. MAIT cell level and function in the peripheral blood were measured by flow cytometry. Circulating MAIT cell levels were found to be significantly reduced in scrub typhus patients. MAIT cell deficiency reflects a variety of clinical conditions. In particular, MAT cell levels reflect disease severity. MAIT cells in scrub typhus patients displayed impaired tumor necrosis factor (TNF)-α production, which was restored during the remission phase. In addition, the impaired production of TNF-α by MAIT cells was associated with elevated CD69 expression.</p><p>Conclusions</p><p>This study shows that circulating MAIT cells are activated, numerically deficient, and functionally impaired in TNF-α production in patients with scrub typhus. These abnormalities possibly contribute to immune system dysregulation in scrub typhus infection.</p></div

Changes in MAIT cell levels and functions in scrub typhus patients.

<p>The percentages of MAIT cells (<u>panel A</u>) in the peripheral blood of 18 scrub typhus patients during active disease and remission were determined by flow cytometry. TNF-α expression (<u>panel B</u>) in the MAIT cell population after stimulation with PMA and IM was determined by intracellular flow cytometry. Data in <u>panel B</u> were obtained from 11 patients with scrub typhus. Symbols represent individual subjects. *p < 0.05 by the Wilcoxon matched-pairs signed rank test.</p

Expression of CD69 and PD-1 and apoptosis of MAIT cells after stimulation with IL-12 and IL-18.

<p>PBMCs (1 × 10⁶/well) were incubated for 24 hours in the presence of IL-12 (50 ng/mL) and IL-18 (50 ng/mL), and then stained with FITC-conjugated anti-CD3, FITC-conjugated annexin V, APC-conjugated anti-TCR Vα7.2, PE-conjugated anti-CD3, PE-conjugated anti-CD69, PE-conjugated anti-PD-1 and PE-Cy5-conjugated anti-CD161 monoclonal antibodies. Percentages of CD69-expressing cells (<u>panel A</u>), annexin V-positive cells (<u>panel C</u>), and PD-1-expressing cells (<u>panel E</u>) among MAIT cells were determined by flow cytometry. Data in <u>panels B, D and F</u> were obtained from 6 HCs. Values are expressed as the mean ± SEM. *p < 0.005 by paired t test.</p

Regression coefficients of MAIT cell percentages with respect to clinical and laboratory parameters in scrub typhus patients.

<p>Regression coefficients of MAIT cell percentages with respect to clinical and laboratory parameters in scrub typhus patients.</p

Highly Enantioselective Phase-Transfer Catalytic Alkylation in the Preparation of Non-natural α-Amino Acids via Solid Phase Synthesis Using Aldimine Linker

A new Merrifield-resin-derived glycinimine tert-butyl ester (9) was prepared and applied to the enantioselective synthesis of non-natural α-amino acids. High enantioselectivities (86 to >99% ee) were accomplished by employing the aldimine linker under phase-transfer alkylation conditions, using 50% aqueous CsOH in toluene/chloroform (7:3) at 0 °C in the presence of N-(9-anthracenylmethyl)-O(9)-allylcinchonidium bromide (10 mol %)

Expression of CD69 and PD-1 and apoptosis of MAIT cells from scrub typhus patients.

<p>Freshly isolated PBMCs were stained with FITC-conjugated anti-CD3, FITC-conjugated annexin V, APC-conjugated anti-TCR Vα7.2, PE-conjugated anti-CD3, PE-conjugated anti-CD69, PE-conjugated anti-PD-1 and PE-Cy5-conjugated anti-CD161 monoclonal antibodies, and then analyzed by flow cytometry. Percentages of CD69-expressing cells (<u>panel A</u>), annexin V-positive cells (<u>panel C</u>) and PD-1-expressing cells (<u>panel E</u>) among MAIT cells were determined by flow cytometry. Data in <u>panels B, D and F</u> were obtained from 14 HCs and 17 patients with scrub typhus. Symbols represent individual subjects and horizontal lines are median values. *p < 0.0001 by the Mann-Whitney U test.</p

Expression of IFN-γ, IL-17 and TNF-α in MAIT cells from scrub typhus patients.

<p>Freshly isolated PBMCs (1 × 10⁶/well) were incubated for 4 hours in the presence of PMA (100 ng/ml) and IM (1 μM). Panel A: Representative IFN-γ, IL-17 and TNF-α expression in the MAIT cell population were determined by intracellular flow cytometry after stimulation with PMA and IM. Data in <u>panel B</u> were obtained from 14 HCs and 23 patients with scrub typhus. Symbols represent individual subjects and horizontal lines are median values. *p < 0.05 by the Mann-Whitney U test.</p