Subjects: pre-season time point (normalized to fold-change

Subjects:          Ten female student girls from the faculty of physicaleducation (Grade 2) constituted subjects of this study. Written informedconsent was obtained from all participants according to the ethical approvalfor this study conformed to the standards of the Declaration of Helsinki.  The Helwan University review board approvedthe protocol before study initiation. Height, weight and age were recorded forall investigated subjects. All participants were required to abstain fromnon-steroidal anti-inflammatory use for at least 7 days prior the experimentalstudy day. Training protocol:The subjects trainedfor 12 weeks as 3 days/week in a basketball program. Each training sessionconsisted of 10 min. warm up followed by 60 – 80 min.

of specific basketballskills training program under specialized coach supervision, then positive cooldown for another 10 min. Exercise on a cycle ergometer to exhaustion wasperformed on the sampling days. Plasma sampling:          Five milliliters of blood was collected under asepticconditions in standard anticoagulant (EDTA)-treated vacutainer tubes atbaseline (prior to exercise testing) and immediately post-exercise testing.

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Allblood samples were centrifuged at 2000 g for 10 min to pellet cellular elementsimmediately after each blood draw. The supernatant plasma was then separatedand immediately frozen at –80?C till PCR assay. miRNA extraction:          TotalRNA extraction was performed using a MicroRNA Extraction Kit (ShanghaiShineGene Molecular Biotech, Inc., Shanghai, China).

Quantitative efficacy andreproducibility of c-miRNA extraction was confirmed by extraction via serialdilutions. Quantification ofmicroRNA expression:To quantify levels ofselect c-miRNA, standard reverse transcription-quantitative (‘real time’)polymerase chain reaction (RT-QPCR) was utilized. Specifically, reversetranscription was performed to generate cDNA representing levels of maturec-miRNA molecules. Fold-change of miRNA 206species was calculated using the formula specified by the manufacturingcompany. Specifically, in this formula, Ct represents the ‘real time’cycle number at which the increase in miRNA probe fluorescence is exponential.As reference control for miRNA quantification, Ct values were thensubtracted from Ct values obtained from exogenously added miR-206, asdescribed above.

Ct values were then compared (?Ct) with each athlete’sown resting baseline at the pre-season time point (normalized to fold-change of1).Basketball skillsMeasurements:Basketball skills were assessed before and after theprogram as follow:A)  Ball defense/obstacleskills for 10 times.B)   Ball passing to/fromanother player at 10 yards during running for 10 times.

C)   Ball shooting to thebasket from the shooting circle for 10 times.D)  Shooting to the basket from the running and jumping position for 10times.Result of total 40 points were calculated andcompared before to after program. Statistical analysis:A Computer program (SPSSversion 17) was used to analyze obtained results. Student’s Mean, standard deviation ofplayers and t test were used to examinethe difference between baseline and after exercise measurements before andafter the training program. Probabilitywas assigned significant at ? 0.05.


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