Characteristics and gene expressions before and after weight loss in obese patients (1^st cohort).

<p>Data shown are means ± SEM.</p><p>*<i>P</i><0.05,</p><p>**<i>P</i><0.01 and</p><p>***<i>P</i><0.001 obese compared with lean controls;</p>$<p><i>P</i><0.05,</p>$$<p><i>P</i><0.01 and</p>$$$<p><i>P</i><0.001 compared with before weight loss;</p><p>Abbreviations: BMI, body mass index; C, cholesterol; DBP, diastolic blood pressure; HOMA-IR, homeostasis model assessment of insulin resistance; hs-CRP, high sensitivity C-reactive protein; ox-LDL, oxidized LDL; SBP, systolic blood pressure.</p

Regulation of IRAK3 expression in THP-1 monocytes.

<p>(<b>A</b>) Gene expression was analyzed by measuring relative <i>RNA</i> levels using qRT-PCR, protein expression and ROS production were determined by flow cytometry in THP-1 cells exposed to 1 or 10 µg/ml gADIPOQ (n = 6) or in <i>IRAK3</i>-depleted THP-1 cells exposed to 10 µg/ml gADIPOQ (n = 4) for 6 h and 24 h. Data shown are means ± SEM of 24 h exposed cells normalized to 6 h exposed cells. ^*<i>P</i><0.05, ^**<i>P</i><0.01 and ^***<i>P</i><0.001 compared with THP-1 cells exposed to high gADIPOQ; ^$$<i>P</i><0.01 and ^$$$<i>P</i><0.001 compared with THP-1 cells exposed to low gADIPOQ. (<b>B</b>) Gene/protein expression in THP-1 cells exposed to 10 µg/ml gADIPOQ and 10 µg/ml ox-LDL (n = 6), 1 µg/ml gADIPOQ and 25 µg/ml ox-LDL (n = 6) or 10 µg/ml gADIPOQ and 25 µg/ml ox-LDL (n = 6). Data are expressed as means ± SEM. ^**<i>P</i><0.01 compared with THP-1 cells exposed to 10 µg/ml gADIPOQ and 10 µg/ml ox-LDL; ^$<i>P</i><0.05 and ^$$<i>P</i><0.01 compared with THP-1 cells exposed to 1 µg/ml gADIPOQ and 25 µg/ml ox-LDL. Abbreviations: gADIPOQ, globular adiponectin, iROS, intracellular ROS; mROS, mitochondrial ROS; ox-LDL, oxidized LDL; ROS, reactive oxygen species.</p

Characteristics of obese women with and without T2DM for validation (2^nd cohort).

<p>Data shown are means ± SEM.</p><p>*<i>P</i><0.05,</p><p>**<i>P</i><0.01 and</p><p>***<i>P</i><0.001 obese compared with lean controls;</p>$<p><i>P</i><0.05,</p>$$<p><i>P</i><0.01 and</p>$$$<p><i>P</i><0.001 compared with obese women without T2DM;</p><p>Abbreviations: BMI, body mass index; C, cholesterol; DBP, diastolic blood pressure; HOMA-IR, homeostasis model assessment of insulin resistance; hs-CRP, high sensitivity C-reactive protein; ox-LDL, oxidized LDL; SBP, systolic blood pressure; T2DM, type 2 diabetes.</p

Gene expressions of the <i>IRAK3</i>-related pathway and adipocyte differentiation markers in visceral adipose tissue.

<p>(<b>A</b>) Gene expression in visceral adipose tissue was analyzed by measuring relative <i>RNA</i> levels using qRT-PCR for key molecules in the TLR2/NFκB inflammatory pathway. The adipose tissue specific antioxidant gene <i>SOD3</i> instead of <i>SOD2</i> was used as oxidative stress marker in visceral adipose tissue. (<b>B</b>) Relative <i>RNA</i> levels of markers of adipocyte differentiation (<i>PPARs</i> and <i>ADIPOQ</i>), insulin signaling (<i>INSR</i>) and glucose uptake (<i>GLUT4</i>) in visceral adipose tissue as determined by qRT-PCR. Data shown are means. ^*<i>P</i><0.05 and ^**<i>P</i><0.01 obese persons compared with lean controls; lean controls (n = 7), obese patients (n = 21).</p

Association of <i>RNA</i> expressions in monocytes and blood levels with occurrence of metabolic syndrome.

<p>*Cut points were determined by ROC curve analysis. Data are means (and 95% confidence intervals). Abbreviations: OR, odds ratio; NPV, negative predictive value; PPV, positive predictive value.</p

Exposure of IRAK3-depleted THP-1 cells to additional stress results in more inflammation and ROS.

<p>Gene expression was analyzed using qRT-PCR and mROS production was determined by flow cytometry in THP-1 cells exposed to (<b>A</b>) 5.5 mM D-glucose and 9.5 mM D-mannitol (osmotic control) or 15 mM D-glucose (n = 6), and (<b>B</b>) 100 ng/ml IL-6 (n = 6). Data shown are means ± SEM. ^*<i>P</i><0.05, ^**<i>P</i><0.01 and ^***<i>P</i><0.001 compared with THP-1 cells exposed to 5.5 mM D-glucose or PBS vehicle. (<b>C</b>) Gene/protein expression and ROS production in THP-1 cells transiently transfected with siRNA targeting <i>IRAK3</i> (n = 10) or in THP-1 cells exposed to 100 mU/ml glucose oxidase with (n = 4) or without (n = 5) silencing of <i>IRAK3</i>. Data shown are means ± SEM. ^*<i>P</i><0.05, ^**<i>P</i><0.01 and ^***<i>P</i><0.001 compared with THP-1 control cells or THP-1 cells transfected with negative control siRNA; ^$$<i>P</i><0.01 and ^$$$<i>P</i><0.001 compared with THP-1 cells transfected with <i>IRAK3</i> siRNA; ^##<i>P</i><0.01 and ^###<i>P</i><0.001 compared with THP-1 cells exposed to glucose oxidase. Abbreviations: iROS, intracellular ROS; mROS, mitochondrial ROS; ROS, reactive oxygen species.</p

At similar weight loss, dietary composition determines the degree of glycemic improvement in diet-induced obese C57BL/6 mice

<div><p>Background</p><p>Achieving weight loss is the cornerstone of the treatment of the metabolic consequences of obesity, in particular of glucose intolerance.</p><p>Objective</p><p>To determine whether improvement in glucose control depends on dietary macronutrient composition of the diet at identical weight loss.</p><p>Materials and methods</p><p>Twenty-two weeks old diet-induced obese C57BL/6 mice lost weight through caloric restriction on normal chow (R-NC) or high fat diet (R-HF). Control mice were fed normal chow (LEAN) or high fat diet (OBESE) ad libitum. Body weight and composition were assessed after 8 weeks of dietary intervention. Glucose homeostasis was evaluated by intraperitoneal glucose tolerance tests (IPGTT). Epididymal white adipose (eWAT) and hepatic tissues were analyzed by immunohistochemistry and RT-qPCR.</p><p>Results</p><p>By 30 weeks of age, the body weight of the mice on R-NC (31.6±1.7g, mean±SEM) and R-HF (32.3±0.9g) was similar to LEAN mice (31.9±1.4g), while OBESE mice weighed 51.7±2.4g. Glucose tolerance in R-NC was better than in LEAN mice (69% AUC IPGTT, P 0.0168) whereas R-HF mice remained significantly less glucose tolerant (125% AUC IPGTT, P 0.0279 vs LEAN), despite identical weight loss. The eWAT pads and adipocyte size were similar in LEAN and R-NC mice, while the eWAT pad size of R-HF was 180% of R-NC (P < 0.0001) and the average adipocyte size of R-HF mice was 134% of R-NC fed mice (P 0.0285). No LEAN or R-NC mice had hepatic steatosis, in contrast to 28.6% of R-HF mice. Compared to OBESE mice, inflammatory markers were lower in eWAT and liver tissue of R-NC, but not in R-HF mice. Measures of visceral adiposity correlated well with glucose tolerance parameters.</p><p>Conclusions</p><p>In mice, caloric restriction on a normal chow diet improved glucose tolerance significantly more when identical weight loss was achieved on a high fat diet.</p></div

Macronutrient composition.

<p>Macronutrient composition.</p

Correlations between body composition, glucose parameters, epididymal white adipose tissue (eWAT) size, hepatic steatosis and ALT levels.

<p>Correlations between body composition, glucose parameters, epididymal white adipose tissue (eWAT) size, hepatic steatosis and ALT levels.</p

Energy expentidure components as obtained in automated cages for indirect calorimetry.

<p>Respiratory exchange rate (a), oxygen consumption (b), heat production (c) and ambulatory activity (d) (n = 6–8 per group). Data are presented as mean±SEM * P<0.05; ** P<0.01 *** P<0.001 **** P<0.0001.</p