Background: Branched-chain amino acids (BCAAs), essential nutrients including leucine, isoleucine, and valine, serve as a resource for energy production and the regulator of important nutrient and metabolic signals. Recent studies have suggested that dysfunction of BCAA catabolism is associated with the risk of cardiovascular disease. Platelets play an important role in cardiovascular disease, but the functions of BCAA catabolism in platelets remain unknown. Methods: The activity of human platelets from healthy subjects before and after ingestion of BCAAs was measured. Protein phosphatase 2Cm specifically dephosphorylates branched-chain α-keto acid dehydrogenase and thereby activates BCAA catabolism. Protein phosphatase 2Cm–deficient mice were used to elucidate the impacts of BCAA catabolism on platelet activation and thrombus formation. Results: We found that ingestion of BCAAs significantly promoted human platelet activity (n=5; P <0.001) and arterial thrombosis formation in mice (n=9; P <0.05). We also found that the valine catabolite α-ketoisovaleric acid and the ultimate oxidation product propionyl-coenzyme A showed the strongest promotion effects on platelet activation, suggesting that the valine/α-ketoisovaleric acid catabolic pathway plays a major role in BCAA-facilitated platelet activation. Protein phosphatase 2Cm deficiency significantly suppresses the activity of platelets in response to agonists (n=5; P <0.05). Our results also suggested that BCAA metabolic pathways may be involved in the integrin αIIbβ3–mediated bidirectional signaling pathway that regulates platelet activation. Mass spectrometry identification and immunoblotting revealed that BCAAs enhanced propionylation of tropomodulin-3 at K255 in platelets or Chinese hamster ovary cells expressing integrin αIIbβ3. The tropomodulin-3 K255A mutation abolished propionylation and attenuated the promotion effects of BCAAs on integrin-mediated cell spreading, suggesting that K255 propionylation of tropomodulin-3 is an important mechanism underlying integrin αIIbβ3–mediated BCAA-facilitated platelet activation and thrombosis formation. In addition, the increased levels of BCAAs and the expression of positive regulators of BCAA catabolism in platelets from patients with type 2 diabetes mellitus are significantly correlated with platelet hyperreactivity. Lowering dietary BCAA intake significantly reduced platelet activity in ob/ob mice (n=4; P <0.05). Conclusions: BCAA catabolism is an important regulator of platelet activation and is associated with arterial thrombosis risk. Targeting the BCAA catabolism pathway or lowering dietary BCAA intake may serve as a novel therapeutic strategy for metabolic syndrome–associated thrombophilia.

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Background Low energy availability (LEA) is a medical condition observed in athletes, with a higher prevalence in aesthetic sports. For the first time, this study evaluated the relative prevalence of LEA in female elite athletes (ELA) and recreational athletes (REA) in aesthetic sports in China. Methods Female athletes from 6 sports (trampolining, rhythmic gymnastics, aerobics, dance sport, cheerleading and dance) were recruited, including ELA (n = 52; age = 20 ± 3) on Chinese national teams and REA (n = 114; Age = 20 ± 2) from Beijing Sport University. Participants completed 2 online questionnaires to assess LEA and eating disorder risk. These included the Low Energy Availability in Females Questionnaire (LEAF-Q), which provided information on injury history, gastrointestinal function and menstrual history, and the Eating Disorder Inventory-3 Referral Form (EDI-3 RF). For a sub-group of elite athletes (n = 14), body composition, bone mineral density, and blood serum were also quantified. Results A total of 41.6% of participants (n = 69) were at increased risk of LEA, and 57.2% of participants (n = 95) were classified as high in eating disorder risk. For ELA vs. REA, there was a significantly higher prevalence of LEA risk (55.8% vs. 35.1%; p = 0.012) and amenorrhea (53.8% vs. 13.3%; p < 0.001). Elite athletes at increased risk of LEA had significantly lower estradiol (p = 0.021) and whole-body BMD (p = 0.028). Pearson correlations indicated that the whole-body BMD (r = − 0.667, p = 0.009) correlated negatively with LEAF-Q score. Conclusions Results of this study indicate that there is a risk of LEA in female Chinese athletes within aesthetic sports, and significantly higher prevalence of increased LEA risk observed in ELA than in REA. Chinese coaches and sports medicine staff working elite female athletes in aesthetic sports should develop strategies to reduce the prevalence of LEA.

Cellular toxicity test is a key step in assessing the graphene toxicity for its biomedical applications. In this study, we investigated the cytotoxicity of graphene with 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2Htetrazolium bromide (MTT) and tetrazolium-8-[2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4disulfophenyl)-2H-tetrazolium] monosodium salt (CCK-8) assay on HepG2 cell line and Chang liver cell line. The cell viability data obtained by using MTT and CCK-8 assay showed inconsistent. Graphene induced adsorption, optical interferences, as well as electron transfer can prevent to appropriate evaluate graphene toxicity. Our findings demonstrated the importance of careful interpreting of obtained data from classical in vitro assays on assessment of graphene cytotoxicity. www.rsc.org/advances 53240 | RSC Adv., 2015, 5, 53240-53244 This journal is