Fucoxanthinol,Metabolite of Fucoxanthin,Improves Obesity-Induced Inflammation in Adipocyte Cells

Fucoxanthin (Fx) is a marine carotenoid found in edible brown seaweeds. We previously reported that dietary Fx metabolite into fucoxanthinol (FxOH), attenuates the weight gain of white adipose tissue of diabetic/obese KK-A^y mice. In this study, to evaluate anti-diabetic effects of Fx, we investigated improving the effect of insulin resistance on the diabetic model of KK-A^y mice. Furthermore, preventing the effect of FxOH on low-grade chronic inflammation related to oxidative stress was evaluated on 3T3-L1 adipocyte cells and a RAW264.7 macrophage cell co-culture system. A diet containing 0.1% Fx was fed to diabetic model KK-A^y mice for three weeks, then glucose tolerance was observed. Fx diet significantly improved glucose tolerance compared with the control diet group. In in vitro studies, FxOH showed suppressed tumor necrosis factor-α (TNF-α), and monocyte chemotactic protein-1 (MCP-1) mRNA expression and protein levels in a co-culture of adipocyte and macrophage cells. These findings suggest that Fx ameliorates glucose tolerance in the diabetic model mice. Furthermore, FxOH, a metabolite of Fx, suppresses low-grade chronic inflammation in adipocyte cells.     

六堡茶对胰岛素抵抗3T3-L1脂肪细胞糖脂代谢的影响

采用软脂酸诱导3T3-L1脂肪细胞胰岛素抵抗后,给予不同剂量的六堡茶水提物治疗,探讨六堡茶对胰岛素抵抗3T3-L1脂肪细胞的糖脂代谢的影响。结果表明,1 mmol·L^-1的软脂酸对3T3-L1脂肪细胞作用24 h后,葡萄糖摄取量减少,建立胰岛素抵抗模型。六堡茶对模型细胞干预24 h后,与模型组比较,细胞培养基的残留液中葡萄糖和游离脂肪酸的含量降低;对糖脂代谢相关关键酶（己糖激酶、6-磷酸果糖激酶-1、丙酮酸激酶、甘油-3-磷酸酰基转移酶）和葡萄糖转运子基因表达都有上调作用,并下调乙酰辅酶A羧化酶、蛋白酪氨酸磷酸酯酶1B基因表达。低剂量的六堡茶下调肉碱脂酰转移酶-I基因的表达,但是高剂量能显著上调肉碱脂酰转移酶-I基因表达。实验证明了六堡茶能够提高胰岛素抵抗3T3-L1脂肪细胞对葡萄糖的摄取能力,同时促进细胞内糖脂代谢,六堡茶有改善3T3-L1脂肪细胞胰岛素抵抗的作用。     

茶桑混合袋泡茶降血糖作用的实验研究

对茶桑混合袋泡茶的降血糖作用进行了研究。以低、中、高三个剂量(0.5、1、3g/kg·bw)的袋泡茶饲喂正常小鼠模型、高血糖小鼠模型14d后,给予葡萄糖2.5g/kg·bw灌胃,于0、0.5h和2.0h时相分别测定其空腹血糖(FBG)和耐糖量(GT)。结果表明,在由四氧嘧啶引起的高血糖模型动物糖耐量试验中,中、高剂量组显著降低了0.5h和2h时相血糖含量和0.5~0h时相血糖差值,且不影响正常对照组的血糖、糖耐量、糖耐量差值,表明茶桑混合袋泡茶具有降低血糖的作用。     

Fucoidan from marine brown algae inhibits lipid accumulation

In this study, we elucidated the inhibitory effect of fucoidan from marine brown algae on the lipid accumulation in differentiated 3T3-L1 adipocytes and its mechanism. The treatment of fucoidan in a dose-dependent manner was examined on lipid inhibition in 3T3-L1 cells by using Oil Red O staining. Fucoidan showed high lipid inhibition activity at 200 μg/mL concentration (P < 0.001). Lipolytic activity in adipocytes is highly dependent on hormone sensitive lipase (HSL), which is one of the most important targets of lipolytic regulation. Here, we examined the biological response of fucoidan on the protein level of lipolysis pathway. The expressed protein levels of total hormone sensitive lipase (HSL) and its activated form, phosphorylated-HSL were significantly increased at concentration of 200 μg/mL fucoidan. Furthermore, insulin-induced 2-deoxy-D-[3H] glucose uptake was decreased up to 51% in fucoidan-treated cells as compared to control. Since increase of HSL and p-HSL expression and decrease of glucose uptake into adipocytes are known to lead to stimulation of lipolysis, our results suggest that fucoidan reduces lipid accumulation by stimulating lipolysis. Therefore, these results suggest that fucoidan can be useful for the prevention or treatment of obesity due to its stimulatory lipolysis.     

茯茶水提物对Ⅱ型糖尿病小鼠糖代谢紊乱的干预作用

采用高脂高糖饲料联合链脲佐菌素诱导Ⅱ型糖尿病小鼠模型,研究茯砖茶水提物对Ⅱ型糖尿病小鼠的降糖功效。结果表明,茯茶水提取物干预能显著改善糖尿病小鼠多食、多饮、多尿、消瘦症状,降低空腹血糖浓度,提高机体对糖负荷的耐受性,提升胰岛素和胰岛素敏感水平,降低胰岛素抵抗水平。同时,q RT-PCR分析发现,茯茶水提物通过增强PPAR-α、GLUT2和GLUT4基因表达,促进了葡萄糖转运和肝脏内脂质代谢,从而改善糖尿病小鼠体内糖代谢紊乱。综上所述,茯茶能显著改善Ⅱ型糖尿病小鼠糖代谢紊乱症状,其中中、高剂量茯茶效果最为显著。     

茶多酚防治2型糖尿病的分子机理研究进展

茶多酚作为天然功能性物质能有效防治2型糖尿病,但是其具体作用机制仍不太清楚。研究表明它的作用机理主要是通过调控糖代谢平衡：包括降低α-糖苷酶、α-淀粉酶等双糖酶活性,调控胰岛素信号传导途径及AMPK途径中多个分子靶点的磷酸化水平,影响酶与转录因子的活性与表达,减弱胰岛素抵抗及糖异生作用,促进胰岛素的合成与分泌,促进靶组织对葡萄糖的吸收与利用等;同时,茶多酚能够调控脂代谢,通过抑制脂质合成与积累的相关酶的活性及转录因子的表达,促进脂质的氧化代谢,减轻脂代谢紊乱;另外,茶多酚的抗氧化与抗炎作用可以有效减轻组织细胞的氧化伤害和炎症损伤,减少细胞凋亡。     

Extensive Screening for Plant Foodstuffs in Okinawa,Japan with Anti-Obese Activity on Adipocytes <Emphasis Type="Italic">In Vitro</Emphasis>

To search for plant foodstuffs with potent anti-obese activity, we conducted a large scale screening based on the inhibitory activity on adipogenesis and the facilitating activity on adipolysis in vitro. That is, inhibition of intracellular lipid accumulation and facilitation of lipid degradation in 3T3-L1 adipocytes were extensively screened from ethanol and hexane extracts of approximately 100 kinds of plant foodstuffs marketed in Okinawa prefecture, which has been famous for the highest prevalence of exceptionally long-lived individuals in the world. Among them thirty one foodstuffs showed potent inhibitory activity on intracellular lipid accumulation in 3T3-L1 adipocytes, whereas only four foodstuffs showed clear facilitating effect on lipid degradation in 3T3-L1 adipocytes. Although further study to examine the in vivo effects on adipogenesis and adipolysis is required, this is the first study to investigate anti-obese characteristics of wide range of traditional Okinawa foodstuffs so that the results give useful information to take another look at Okinawa food culture.     

Ameliorating Effects of Fermented Rice Bran Extract on Oxidative Stress Induced by High Glucose and Hydrogen Peroxide in 3T3-L1 Adipocytes

In this study, we investigated whether fermented rice bran (FRB) can ameliorate the oxidative stress induced by high glucose and hydrogen peroxide (H₂O₂) in 3T3-L1 adipocytes by analyzing reactive oxygen species (ROS), oil red O staining, as well as the expression of mRNAs related to glucose homeostasis and adipogenesis. It was first confirmed that rice bran fermented by Issatchenkia orientalis MFST1 extract increased free phenolic content compared to non-fermented rice bran. The FRB extract strongly inhibited ROS generation and upregulated the expression of PPAR-γ and adiponectin. Moreover, FRB upregulated GLUT4 related to glucose transportation and insulin sensitivity. Taken together, FRB extract ameliorated oxidative stress-induced insulin resistance by neutralizing free radicals and upregulating adiponectin in adipocytes. Our results provide information toward understanding the beneficial effects of FRB on oxidative stress.     

Comparative Study of Sargassum fusiforme Polysaccharides in Regulating Cecal and Fecal Microbiota of High-Fat Diet-Fed Mice

Seaweed polysaccharides represent a kind of novel gut microbiota regulator. The advantages and disadvantages of using cecal and fecal microbiota to represent gut microbiota have been discussed, but the regulatory effects of seaweed polysaccharides on cecal and fecal microbiota, which would benefit the study of seaweed polysaccharide-based gut microbiota regulator, have not been compared. Here, the effects of two Sargassum fusiforme polysaccharides prepared by water extraction (SfW) and acid extraction (SfA) on the cecal and fecal microbiota of high-fat diet (HFD) fed mice were investigated by 16S rRNA gene sequencing. The results indicated that 16 weeks of HFD dramatically impaired the homeostasis of both the cecal and fecal microbiota, including the dominant phyla Bacteroidetes and Actinobacteria, and genera Coriobacteriaceae, S24-7, and Ruminococcus, but did not affect the relative abundance of Firmicutes, Clostridiales, Oscillospira, and Ruminococcaceae in cecal microbiota and the Simpson’s index of fecal microbiota. Co-treatments with SfW and SfA exacerbated body weight gain and partially reversed HFD-induced alterations of Clostridiales and Ruminococcaceae. Moreover, the administration of SfW and SfA also altered the abundance of genes encoding monosaccharide-transporting ATPase, α-galactosidase, β-fructofuranosidase, and β-glucosidase with the latter showing more significant potency. Our findings revealed the difference of cecal and fecal microbiota in HFD-fed mice and demonstrated that SfW and SfA could more significantly regulate the cecal microbiota and lay important foundations for the study of seaweed polysaccharide-based gut microbiota regulators.     

Attenuating Effects of Dieckol on High-Fat Diet-Induced Nonalcoholic Fatty Liver Disease by Decreasing the NLRP3 Inflammasome and Pyroptosis

Nonalcoholic fatty liver disease (NAFLD), which promotes serious health problems, is related to the increase in the nucleotide-binding oligomerization domain-like receptor family, pyrin domain containing 3 (NLRP3) inflammasome and pyroptosis by a high-fat diet (HFD). Whether dieckol (DK), a component of Ecklonia cava extracts (ECE), attenuated NAFLD in an HFD-induced NAFLD animal model was evaluated. The expression of high mobility group box 1/Toll-like receptor 4/nuclear factor-κB, which initiated the NLRP3 inflammasome, was increased in the liver of HFD-fed animals and significantly decreased with ECE or DK administration. The expression of NLRP3/ASC/caspase-1, which are components of the NLRP3 inflammasome, and the number of pyroptotic cells were increased by HFD and decreased with ECE or DK administration. The accumulation of triglycerides and free fatty acids in the liver was increased by HFD and decreased with ECE or DK administration. The histological NAFLD score was increased by HFD and decreased with ECE or DK administration. The expression of lipogenic genes (FASN, SREBP-2, PPARγ, and FABP4) increased and that of lipolytic genes (PPARα, CPT1A, ATGL, and HSL) was decreased by HFD and attenuated with ECE or DK administration. In conclusion, ECE or DK attenuated NAFLD by decreasing the NLRP3 inflammasome and pyroptosis.     

Fucoidan and Fucoxanthin Attenuate Hepatic Steatosis and Inflammation of NAFLD through Modulation of Leptin/Adiponectin Axis

Non-alcoholic fatty liver disease (NAFLD) is the emerging cause of chronic liver disease globally and lack of approved therapies. Here, we investigated the feasibility of combinatorial effects of low molecular weight fucoidan and high stability fucoxanthin (LMF-HSFx) as a therapeutic approach against NAFLD. We evaluated the inhibitory effects of LMF-HSFx or placebo in 42 NAFLD patients for 24 weeks and related mechanism in high fat diet (HFD) mice model and HepaRG^TM cell line. We found that LMF-HSFx reduces the relative values of alanine aminotransferase, aspartate aminotransferase, total cholesterol, triglyceride, fasting blood glucose and hemoglobin A1c in NAFLD patients. For lipid metabolism, LMF-HSFx reduces the scores of controlled attenuation parameter (CAP) and increases adiponectin and leptin expression. Interestingly, it reduces liver fibrosis in NAFLD patients, either. The proinflammatory cytokines interleukin (IL)-6 and interferon-γ are reduced in LMF-HSFx group. In HFD mice, LMF-HSFx attenuates hepatic lipotoxicity and modulates adipogenesis. Additionally, LMF-HSFx modulates SIRI-PGC-1 pathway in HepaRG cells under palmitic acid-induced lipotoxicity environment. Here, we describe that LMF-HSFx ameliorated hepatic steatosis, inflammation, fibrosis and insulin resistance in NAFLD patients. LMF-HSFx may modulate leptin-adiponectin axis in adipocytes and hepatocytes, then regulate lipid and glycogen metabolism, decrease insulin resistance and is against NAFLD.     

Effect of N. sativa oil on impaired glucose tolerance and insulin insensitivity induced by high-fat-diet and turpentine-induced trauma

The aim of this study was to investigate the effect of N. sativa oil on impaired glucose tolerance and insulin insensitivity induced by high-fat diet and trauma. Three dietary groups were used in this study; Rat-Chow (RC), N. sativa oil diet (Combination 4% N. sativa oil and 16% butter oil) (NSOD) and 20% Butter Oil Diet (BOD). Each group was subdivided in two groups; control and trauma. Diets were supplemented for five consecutive weeks body weight increase per week was calculated. At end of the dietary treatments, single dose (2 mL kg(-1) body weight) of turpentine was injected in the dorso-lumber region. Intravenous glucose tolerance test (i.v. GTT) was performed, insulinogenic index and insulin sensitivity was measured. The results showed butter oil diet significantly increased the body weights and visceral fats compared other two groups, respectively. Fasting glucose levels did not change in trauma induced rats while insulin levels increased significantly and it found highest in butter oil diet fed animals. Impaired glucose tolerance was found sever in BOD fed traumatized rats. N. sativa oil diet protected impaired glucose tolerance and insulin insensitivity induced either via saturated fatty acids or injury. In conclusion, N. sativa oil may be used in post surgery diabetic patients to prevent the long going adverse effects from surgical trauma.     

Dieckol Decreases Caloric Intake and Attenuates Nonalcoholic Fatty Liver Disease and Hepatic Lymphatic Vessel Dysfunction in High-Fat-Diet-Fed Mice

Increased inflammation is the main pathophysiology of nonalcoholic fatty liver disease (NAFLD). Inflammation affects lymphatic vessel function that contributes to the removal of immune cells or macromolecules. Dysfunctional lymphatic vessels with decreased permeability are present in NAFLD. High-fat diet (HFD) is known to increase body weight, food intake, and inflammation in the liver. Previously, it was reported that Ecklonia cava extracts (ECE) decreased food intake or weight gain, and low-calorie diet and weight loss is known as a treatment for NAFLD. In this study, the effects of ECE and dieckol (DK)—which is one component of ECE that decreases inflammation and increases lymphangiogenesis and lymphatic drainage by controlling lymphatic permeability in high-fat diet (HFD)-fed mice—on weight gain and food intake were investigated. ECE and DK decreased weight gain and food intake in the HFD-fed mice. NAFLD activities such as steatosis, lobular inflammation, and ballooning were increased by HFD and attenuated by ECE and DK. The expression of inflammatory cytokines such as IL-6 and TNF-α and infiltration of M1 macrophages were increased by HFD, and they were decreased by ECE or DK. The signaling pathways of lymphangiogenesis, VEGFR-3, PI3K/pAKT, and pERK were decreased by HFD, and they were restored by either ECE or DK. The expression of VE-cadherin (which represents lymphatic junctional function) was increased by HFD, although it was restored by either ECE or DK. In conclusion, ECE and DK attenuated NAFLD by decreasing weight gain and food intake, decreasing inflammation, and increasing lymphangiogenesis, as well as modulating lymphatic vessel permeability.     

A New Fungal Triterpene from the Fungus Aspergillus flavus Stimulates Glucose Uptake without Fat Accumulation

Through activity-guided fractionation, a new triterpene (asperflagin, 1) was isolated as a PPAR-γ agonist from the jellyfish-derived fungus Aspergillus flavus. Asperflagin displayed selective and moderate transactivation effects on PPAR-γ in Ac2F rat liver cells. Based on further biological evaluation and molecular docking analysis, we postulated that asperflagin might function as a PPAR-γ partial agonist. This compound was calculated to display a typical PPAR-γ ligand–receptor interaction that is distinct from that of full agonistic antidiabetics such as rosiglitazone, and may retain the antidiabetic effect without accompanying weight gain. Weight gain and obesity are typical side effects of the PPAR-γ full agonist rosiglitazone, and lead to suboptimal outcomes in diabetic patients. Compared to rosiglitazone, asperflagin showed higher glucose uptake in HepG2 human liver cells at concentrations of 20 and 40 μM but induced markedly lower adipogenesis and lipid accumulation in 3T3-L1 preadipocytes. These results suggest that asperflagin may be utilized for further study on advanced antidiabetic leads.     

Antihyperglycemic Effects of <Emphasis Type="Italic">Platycodon grandiflorum</Emphasis> (Jacq.) A. DC. Extract on Streptozotocin-induced Diabetic Mice

The root of Platycodon grandiflorum (Jacq.) A. DC has been reported to have a wide range of health benefits in oriental food. This study examined the hypoglycemic effects of Platycodon grandiflorum (Jacq.) A. DC aqueous-ethanol extract (PGE) in streptozotocin (STZ) -induced diabetic ICR mice (STZ diabetic mice) for the first time. The effects of PGE on blood glucose, plasma insulin levels and body weight were investigated. A significant decrease in blood glucose levels was observed after single administration of PGE. Furthermore, Glibenclamide and PGE significantly suppressed the rise in blood glucose after 30 min in the acute glucose tolerance test. Treatment with glibenclamide and PGE resulted in a reduction in blood glucose levels from the 2nd week, and this reduction was maintained until the 4th week of treatment. The body weight changed slightly in glibenclamide and PGE treated mice in comparison with the STZ control group. Plasma insulin levels were increased with glibenclamide treatment in STZ diabetic mice, whereas such effect was not observed with PGE. These results indicated that PGE could induce hypoglycemic effects without stimulating insulin secretion.     

Beta cell protective effects of sodium tungstate in streptozotocin-induced diabetic rats: glycemic control, blockage of oxidative stress and beta cell histochemistry

Background: Diabetes is a major public health problem. The development of new therapies that are able to improve glycemia management and even to cure diabetes is of great interest. In this study, protective effects of sodium tungstate against STZ-induced beta-cell damages were investigated. Methods: Sixty rats were divided into six groups: control, diabetic, sodium tungstate treated diabetic rats from one week before STZ injection (TDB), food-restricted diabetic (FRD), tungstate treated control, sodium tungstate treated diabetic rats from one week after STZ administration (TDA). We evaluated serum insulin, glucose and glucose tolerance; liver glycogen content, glucokinase (GK) activity; blood and pancreas antioxidant power, lipid peroxidation; and fuchsin-aldehyde histochemical staining of beta-cells. Results: Blood glucose levels of TDB group were lower than other diabetic groups (P<0.01). Blood insulin levels of all diabetic groups were lower than controls (P<0.01). Glucose intolerance improved in TDB animals. Blood and pancreas antioxidant power, liver glycogen contents and GK activities and granulated beta cells increased in TDB rats in comparison with other diabetic groups (P<0.01). Likewise, lipid peroxidation decreased significantly in TDB rats (P<0.01). Conclusions: Results suggested that sodium tungstate if administrated before STZ injection improves glycemic state by a direct effect on pancreatic beta-cells and preserves them by reducing the activity of these cells at the time of STZ injection, reducing STZ-induced oxidative stress, reducing insulin secretion, or all of the above mentioned.     

安吉白茶多糖对实验性糖尿病小鼠的降血糖作用研究

通过制备去甲肾上腺素致糖尿病小鼠模型及四氧嘧啶致高血糖小鼠模型,给正常小鼠、去甲肾上腺素致高血糖小鼠及四氧嘧啶致糖尿病小鼠连续灌胃安吉白茶多糖14 d后,取血测定血糖水平,以探讨安吉白茶多糖对正常小鼠、去甲肾上腺素致高血糖小鼠及四氧嘧啶致高血糖小鼠血糖水平及糖耐量（GT）的影响。结果显示,灌胃2周后,安吉白茶多糖对正常小鼠血糖水平影响较小,低、中、高剂量组与正常对照组相比均无显著意义(P>0.05);安吉白茶多糖能明显降低去甲肾上腺素所致糖尿病小鼠的血糖水平,低、中、高剂量组的血糖值与肾上腺素模型组比较,差异极显著(P<0.01)。且高剂量组降血糖效果优于低剂量组;安吉白茶多糖明显降低四氧嘧啶所致高血糖小鼠的血糖水平,三剂量组的血糖值与四氧嘧啶模型组比较,差异极显著(P<0.01)。三剂量均能降低实验性高血糖小鼠血糖,且随剂量增大,降血糖作用增强,以高剂量降糖作用最强。安吉白茶多糖在缓解糖尿病小鼠糖耐量降低方面作用显著,达到了药物组的治疗水平,但并不影响正常小鼠的血糖和糖耐量。     

Antidiabetic Activity of Differently Regioselective Chitosan Sulfates in Alloxan-Induced Diabetic Rats

The present study investigated and compared the hypoglycemic activity of differently regioselective chitosan sulfates in alloxan-induced diabetic rats. Compared with the normal control rats, significantly higher blood glucose levels were observed in the alloxan-induced diabetic rats. The differently regioselective chitosan sulfates exhibited hypoglycemic activities at different doses and intervals, especially 3-O-sulfochitosan (3-S). The major results are as follows. First, 3,6-di-O-sulfochitosan and 3-O-sulfochitosan exhibited more significant hypoglycemic activities than 2-N-3, 6-di-O-sulfochitosan and 6-O-sulfochitosan. Moreover, 3-S-treated rats showed a more significant reduction of blood glucose levels than those treated by 3,6-di-O-sulfochitosan. These results indicated that –OSO₃⁻ at the C3-position of chitosan is a key active site. Second, 3-S significantly reduced the blood glucose levels and regulated the glucose tolerance effect in the experimental rats. Third, treatment with 3-S significantly increased the plasma insulin levels in the experimental diabetic rats. A noticeable hypoglycemic activity of 3-S in the alloxan-induced diabetic rats was shown. Clinical trials are required in the future to confirm the utility of 3-S.     

Lipid Inhibitory Effect of (−)-loliolide Isolated from Sargassum horneri in 3T3-L1 Adipocytes: Inhibitory Mechanism of Adipose-Specific Proteins

Sargassum horneri (S. horneri) is a well-known brown seaweed widely distributed worldwide. Several biological activities of S. horneri have been reported. However, its effects on lipid metabolism and the underlying mechanisms remain elusive. In the present study, we examined the inhibitory effect of the active compound “(−)-loliolide ((6S,7aR)-6-hydroxy-4,4,7a-trimethyl-5,6,7,7a-tetrahydro-1-benzofuran-2(4H)-one (HTT))” from S. horneri extract on lipid accumulation in differentiated adipocytes. MTT assays demonstrated that (−)-loliolide is not toxic to 3T3-L1 adipocytes in a range of concentrations. (−)-loliolide significantly reduced intracellular lipid accumulation in the differentiated phase of 3T3-L1 adipocytes as shown by Oil Red O staining. Western blot analysis revealed that (−)-loliolide increased the expression of lipolytic protein phospho-hormone-sensitive lipase (p-HSL) and thermogenic protein peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1). Additionally, (−)-loliolide decreased expression of adipogenic and lipogenic proteins, including sterol regulatory element-binding protein-1 (SREBP-1), peroxisome proliferator-activated receptor-γ (PPAR-γ), CCAAT/enhancer-binding protein-α (C/EBP-α), and fatty acid-binding protein 4 (FABP4) in 3T3-L1 adipocytes. These results indicate that (−)-loliolide from S. horneri could suppress lipid accumulation via regulation of antiadipogenic and prolipolytic mechanisms in 3T3-L1 cells. Considering the multifunctional effect of (−)-loliolide, it can be useful as a lipid-lowering agent in the management of patients who suffer from obesity.