ABSTRACTS from 16th International COnference on ... - CRRT Online

<strong>ABSTRACTS</strong> FROM 17 TH INTERNATIONAL CONFERENCE ON CRRT,
SAN DIEGO, FEB 14-17, 2012
second trough blood and effluent
samples were drawn and immediately
stored on ice. Drug analysis: Free and
effluent imipenem levels were measured
by RP-HPLC in the lab of one of the
investigators (WHF). Effluent and free
plasma imipenem levels were compared
using a multivariate linear regression.
Results: Complete data was available
<strong>from</strong> 17 subjects dialyzed with the
NxStage Express (n=6) or Gambro
Prismaflex (n=11). Plasma total
imipenem levels were not measurable
using our HPLC assay, but plasma free
drug and effluent drug levels were
measured in 51 paired samples. Effluent
levels predicted plasma free drug levels
in a CRRT-dose dependent manner.
Effluent overestimated plasma in one
sample. Discussion: Imipenem analysis
in CRRT effluent may provide a bloodsparing
technique for pharmacokinetic
and pharmacodynamic studies. More
study is needed to determine the best use
of this technique.
73. Perfluorocarbon Protects Kidney
Tubular Epithelial Cells By Septic
Plasma-Induced Apoptosis And
Promotes CD133+ Renal
Progenitor Cell Differentiation:
Relevance For Bioartificial Renal
Assist Devices
Vincenzo Cantaluppi, Davide Medica,
Alessandro D Quercia, Federico
Figliolini, Sergio Dellepiane, Gennaro
Iavarone, Giovanni Abagnale, Giuseppe
P Segoloni, Giovanni Camussi
University of Turin, San Giovanni
Battista Molinette Hospital
Extracorporeal blood purification
techniques including renal assist devices
(RAD) with viable renal tubular
epithelial cells (TEC) have been
proposed for the treatment of sepsisassociated
acute kidney injury (AKI).
We previously demonstrated that plasma
derived <strong>from</strong> septic patients induce a
direct injury and pro-apoptotic effect on
cultured human TEC. Perfluorocarbon
(PFC) molecules are oxygen carriers
used for organ preservation before
transplantation. The aim of this study
was to evaluate the effect of PFC on
septic plasma-induced TEC injury and
on renal CD133+ stem cell
differentiation. TEC and CD133+ renal
progenitors were isolated by cell sorting.
Plasma was drawn by 1 patients with
sepsis and AKI (RIFLE criteria). TEC
were incubated with patients’ plasma in
presence or absence of PFC evaluating:
cytotoxicity (XTT assay), apoptosis
(TUNEL assay, ELISA for caspase-3, -8,
-9), cell polarity (trans-epithelial
electrical resistance, TER) and albumin
uptake. Moreover, we studied the effect
of PFC on proliferation (BrdU assay)
and differentiation of CD133+ renal
progenitor cells. Septic plasma induced:
1) a cytotoxic and pro-apoptotic effect
on TEC through the activation of the
death receptor as well as of the
mitochondrial apoptotic pathways; 2) the
alteration of cell polarity (TER) and
albumin uptake; 3) the down-regulation
of the tight junction protein ZO-1 and of
the endocytic receptor megalin. All the
detrimental effects induced by septic
plasma on TEC were significantly
reduced in presence of PFC. In addition,
PFC induced CD133+ progenitor cell
proliferation and differentiation toward
an epithelial phenotype (increase of TER
and expression of markers of fully
differentiated TEC such as E-cadherin,
ZO-1, megalin, alkaline phosphatase,
aminopeptidase-A, aquaporin-1 and
NGAL ). Conclusion: PFC protects
TEC <strong>from</strong> septic plasma-induced injury
and provides an appropriated oxygen
tension to promote CD133+ stem cell
179