CD38 expression have been described to be highly associated in CLL, we sought to determine their functional consequences in the context of supportive BM niches. For this we employed in vivo adoptive transfers of human leukemic cells into immunodeficient mice, in vitro flow cytometrical phenotyping.The steady-state lactate accumulation was attained in 10 minutes, indicating a fast lactate uptake by Bcap-37 cells or a fast equilibrium of lactate concentration across plasma membrane. On the other hand, intracellular pyruvate concentration was not significantly changed. As a result, L/P ratio throughout entire incubation increased by about 8 folds. Given the near-equilibrium status of the conversion in cells with Q value that is smaller but often within one order of magnitude less than Keq, this 8-fold increase of L/P ratio would likely lead the conversion to equilibrium.

In order to test whether the increase of L/P ratio could affect the equilibrium state, we measured glucose consumption and lactate generation by Bcap-37 cells that were incubated in medium containing exogenous lactate. In the absence of exogenous lactate, cells converted quantity glucose to lactate with a high L/G ratio, indicating that the net flow is from pyruvate to lactate. In the presence of exogenous 16 mM lactate, the L/G ratio was dramatically reduced to 0.260.03, indicating that the conversion was approaching equilibrium. In the presence of 19 mM lactate, cells produced negligible lactate and the L/G ratio was close to zero, indicating that the conversion was at or very close to equilibrium. In the presence of 22 mM lactate, cells consumed lactate, indicating that the direction of the conversion was reversed. We then measured the intracellular concentration of lactate and pyruvate, and L/P ratios in cells cultured under above conditions.Despite the fact that functional assays did not show any effect of RNA helicase DHX36 and HuR on cryptic exon inclusion, involvement of these proteins in regulation of cryptic exon activation cannot be completely ruled out as it might be that their effect on processing of ATM intron 20 and aberrant cryptic exon activation is compensated by other proteins that are yet to be identified.

In conclusion, we have identified two splicing factors, hnRNPA1/A2 and DAZAP1, that regulate ATM cryptic exon inclusion in an ISE-dependent manner. Pathological amyloidosis is characterized by the progressive formation in cells and organs of proteinaceous aggregates consisting predominantly of highly ordered, cross-b-sheet fibrils.It is also known from a small study by Valzania et al., that hemodynamic parameters tend to change over the course of several months after CRT. So, it would be very interesting, if these modest changes in EF immediately after implantation translate into robust changes later on in the course. Further follow-up has to clarify this issue.