Comparison of two methods of cardiopulmonary exercise testing for assessing physical fitness in children and adolescents with extreme obesity.

It is well-known that children and adolescents with obesity have increased over recent decades which in turn carries greater risk of co-morbidities and poses a preventive as well as a therapeutic challenge. Currently, there are limited recommendations available on proven methods for recording physical fitness in children and adolescents presenting with extreme obesity. In this study, twenty participants, aged 12-17 years, with a body mass index (BMI) above the 99.5th percentile, were comparatively assessed, using a correlation between their physical fitness on a bicycle (BC) and treadmill (TM) cardiopulmonary exercise testing (CPET) with a lactate diagnostic. The results of the BC and the TM were as follows: maximum heart rate (HRmax) 186.4 ± 8.6 beats per minute (bpm) vs. 190.8 ± 8.8 bpm, peak oxygen consumption (VO2peak/kg) 23.5 ± 2.9 ml/min/kg vs. 25.4 ± 3.1 ml/min/kg, and maximum lactate (Lamax) 6.4 ± 1.6 mmol/l vs. 5.6 ± 1.4 mmol/l. The values of HRmax and VO2peak/kg were significantly higher for adolescents tested on the TM. However, no significant difference was observed in either Lamax values or between the genders.    Conclusions: The higher values of HRmax and VO2peak/kg could be attributed to the activation of a higher percentage of muscle mass on the TM. Lower Lamax values on the TM suggest maximum physical exertion was not achieved. This could be due to the extreme body weight carried by the participants. Both the BC and the TM CPET could be used for assessing physical fitness in children and adolescents with extreme obesity but should not be used interchangeably. What is Known: • Currently, there are only limited recommendations available on proven methods for recording physical fitness in children and adolescents with extreme obesity available. What is New: • Cardiopulmonary exercise testing with maximum physical exertion has been shown to be feasible in children and adolescents with extreme obesity. The results obtained from this study demonstrated that both a bicycle and a treadmill can be effectively used for assessing the physical fitness levels in children and adolescents with extreme obesity.

Two patterns of thrombopoietin signaling suggest no coupling between platelet production and thrombopoietin reactivity in thrombocytopenia-absent radii syndrome.

BACKGROUND: Thrombocytopenia with absent radii syndrome is defined by bilateral radius aplasia and thrombocytopenia. Due to impaired thrombopoietin signaling there are only few bone marrow megakaryocytes and these are immature; the resulting platelet production defect improves somewhat over time. A microdeletion on chromosome 1q21 is present in all patients but is not sufficient to form thrombocytopenia with absent radii syndrome. We aimed to refine the signaling defect in this syndrome. DESIGN AND METHODS: We report an extended study of 23 pediatric and adult patients suffering from thrombocytopenia with absent radii syndrome in order to scrutinize thrombopoietin signal transduction by immunoblotting and gel electrophoretic shift assays. In addition, platelet immunotyping and reactivity were analyzed by flow cytometry. Results were correlated with clinical data including age and platelet counts. RESULTS: Two distinct signaling patterns were identified. Juvenile patients showed abrogated thrombopoietin signaling (pattern #1), which is restored in adults (pattern #2). Phosphorylated Jak2 was indicative of activation of STAT1, 3 and 5, Tyk2, ERK, and Akt, showing its pivotal role in distinct thrombopoietin-dependent pathways. Jak2 cDNA was not mutated and the thrombopoietin receptor was present on platelets. All platelets of patients expressed normal levels of CD41/61, CD49b, and CD49f receptors, while CD42a/b and CD29 were slightly reduced and the fibronectin receptor CD49e markedly reduced. Lysosomal granule release in response to thrombin receptor activating peptide was diminished. CONCLUSIONS: We show a combined defect of platelet production and function in thrombocytopenia with absent radii syndrome. The rise in platelets that most patients have during the first years of life preceded the restored thrombopoietin signaling detected at a much later age, implying that these events are uncoupled and that an unknown factor mediates the improvement of platelet production.

The microtubule modulator RanBP10 plays a critical role in regulation of platelet discoid shape and degranulation.

Terminally mature megakaryocytes undergo dramatic cellular reorganization to produce hundreds of virtually identical platelets. A hallmark feature of this process is the generation of an elaborate system of branched protrusions called proplatelets. We recently identified RanBP10 as a tubulin-binding protein that is concentrated along polymerized microtubules in mature megakaryocytes. RanBP10 depletion in vitro caused the disturbance of polymerized filaments. Here we study the function of RanBP10 in vivo by generating deficient mice using a gene-trap approach. Mutant mice show normal platelet counts, and fetal liver-derived megakaryocytes reveal only slightly reduced proplatelet formation. However, ultrastructural analysis unveiled a significantly increased geometric axis ratio for resting platelets, and many platelets exhibited disorders in microtubule filament numbers and localization. Mutant mice showed a markedly prolonged bleeding time. Granule release, a process that depends on internal contraction of the microtubule marginal coil, also was reduced. Flow cytometry analysis revealed reduced expression of CD62P and CD63 after PAR4-peptide stimulation. These data suggest that RanBP10 plays an essential role in hemostasis and in maintaining microtubule dynamics with respect to both platelet shape and function.