[Effect of 99mTc elution in vivo from red cells on red cell volumes measured using autologous 99mTc-labeled red cells: comparison with 51Cr method]. / Effet de l'élution in vivo du technétium sur les volumes globulaires mesurés à l'aide d'hématies autologues marquées au 99mTc : comparaison avec la technique au 51Cr.

The purpose of this work was to compare the measured red-cell volume (RCV) using sodium pertechnétate [RCV-99mTc] compared to the reference technique using sodium radiochromate [RCV-51Cr] and to assess the influence of technetium-99 elution on the RCV-99mTc value. Ten patients had simultaneous measurements of RCV-99mTc and RCV-51Cr. Elution of Tc-99m from red blood cells was 2.9% and led to an average overestimation of RCV-99mTc of 3.7%. The introduction of individual tracer elution rates in the RCV-99mTc calculation corrects this overestimation.

Significant differences when using MDRD for GFR estimation compared to radionuclide measured clearance.

OBJECTIVES: To evaluate Estimated Glomerular Filtration Rate (eGFR), using the Modification of Diet in Renal Disease equation, and compare with radionuclide GFR (rGFR) in a Radiology setting to assess renal function prior to contrast administration. METHODS: Five hundred and sixteen retrospective rGFR studies from a mixed referral population were selected and the eGFR calculated. Regression and Bland-Altman analysis was performed. The percentage of rGFR and eGFR studies below 30 ml/min/1.73 m² and 60 ml/min/1.73 m² were calculated; these are important thresholds for classifying renal insufficiency. RESULTS: A significant correlation between eGFR and rGFR (R² = 0.62, p < 0.0001) and significant differences in the medians (p < 0.0001) were found. eGFR overestimated rGFR with a bias (mean difference) of 10.8 ml/min/1.73 m² over the whole range of rGFR. Studies with an rGFR of under 30 ml/min/1.73 m² had a mean bias of 4.6 ml/min/1.73 m² (difference range -5.9 to 26.3 ml/min/1.73 m²). The bias over the range 30 to 60 ml/min/1.73 m² was 13.2 ml/min/1.73 m² (difference range -16.8 to 88.3 ml/min/1.73 m²). In 25.4% of studies, eGFR was less than 60 ml/min/1.73 m² compared with 40.5% of rGFR studies. CONCLUSIONS: Awareness of the bias between eGFR and rGFR is important when assessing Radiology patients for risks of nephrotoxicity and Nephrogenic Systemic Fibrosis from contrast medium.

Intestinal permeability in cirrhotic patients with and without spontaneous bacterial peritonitis: is the ring closed?

OBJECTIVES: Impaired intestinal permeability (IP) may have a role in the pathogenesis of ascites and in spontaneous bacterial peritonitis (SBP) in patients with liver cirrhosis (LC). The aim of this study was to assess IP in LC patients with respect to healthy controls. METHODS: IP was evaluated by the (51)Cr-ethylenediaminetetraacetic acid ((51)Cr-EDTA) permeability test in 52 LC patients and in 48 sex- and age-matched controls. The presence of (51)Cr-EDTA was also evaluated in ascitic fluid after therapeutic paracentesis in all LC patients with ascites. RESULTS: An altered IP was found in 45% of LC patients compared with 4% of controls (P<0.00001). IP impairment was significantly associated with Child-Pugh status (75% of Child C patients vs. 39% of Child B and 22% of Child A patients), with the presence of ascites (60% in ascitic patients vs. 31% in nonascitic patients), and with a history of SBP (100% of patients with SBP vs. 50% of those without SBP). (51)Cr-EDTA was present in all ascitic samples obtained from patients with SBP compared with 22% of patients without SBP. CONCLUSIONS: IP derangement was a common finding in LC, especially in patients with more advanced disease (presence of ascites and history of SBP). The presence of (51)Cr-EDTA in ascites in patients with SBP suggests an altered permeability of splancnic vessels and/or peritoneal membranes. Further studies are required to assess (51)Cr-EDTA urine and ascite cutoffs to set up SBP preventive strategies.

Assessment of glomerular filtration rate by serum cystatin C in patients undergoing coronary artery bypass grafting.

BACKGROUND: Assessment of renal function in patients undergoing coronary artery bypass grafting (CABG) is important. Cystatin C has been proposed as an improved indicator of renal function. The aim of this study was to assess cystatin C as an early marker of changes in glomerular filtration rate (GFR) after CABG. METHODS: Blood samples were collected from 61 CABG patients at different time points. Using (51)Cr-ethylenediaminetetraacetic acid ((51)Cr-EDTA) clearance as a 'gold standard', we compared the correlations and non-parametric receiver operator characteristic curves of serum cystatin C, serum creatinine and 24 h creatinine clearance (Ccr). RESULTS: The inverse of cystatin C correlated better with (51)Cr-EDTA than those of serum creatinine and Ccr (r = 0.8578, 0.6771 and 0.6929, respectively). Cystatin C exhibited significantly superior diagnostic accuracy for detecting GFR <80 mL/min/1.73 m(2) compared with serum creatinine (P = 0.013) and Ccr (P = 0.025); for detecting GFR <60 mL/min/1.73 m(2), cystatin C had similar diagnostic accuracy to Ccr (P = 0.812) but was superior to creatinine (P = 0.033). At the best cut-off value, cystatin C had sensitivity 89% and specificity 93% for detecting GFR <80 mL/min/1.73 m(2), sensitivity 86% and specificity 96% for detecting GFR <60 mL/min/1.73 m(2). CONCLUSIONS: Cystatin C is a better marker for detecting small temporary changes of GFR in CABG patients. This may allow better identification of patients with renal impairment.

Checking the consistency of the two blood samples slope-intercept method for estimating GFR using the single blood sample formula in children.

BACKGROUND: The slope-intercept method is widely used for the determination of the plasma clearance of 51Cr-EDTA. When three or more plasma samples are used, the goodness-of-fit (r2) can be used as a measure of consistency of the samples. This parameter can not be used, however, if only two samples are available. PURPOSE: To evaluate whether the single-sample technique (SBS) can be used to check the consistency of the slope-intercept method using two blood samples (2BS) in children. METHODS: Simulated computer models of a mono-exponential curve were created in order to represent three children aged 3, 6 and 10 years, each with a large range of clearances values and three distribution volumes, respectively 20%, 25% and 30% of body weight. Errors were then introduced in the injected dose (errors from -50% to +50%) and on the 120 or 240 min blood sample (errors from -50% to +50%). The effects of these errors on the clearance measurement using 2BS and SBS methods were calculated and compared. RESULTS: The errors on the injected dose, the 120 min and 240 min plasma samples introduced errors in the same direction and with the same magnitude on both the SBS and 2BS clearance values. For that reason, the comparison between the SBS methods and the 2BS techniques has a low sensitivity in detecting an eventual error. Striking differences between the SBS method and the 2BS technique were only observed when considerable errors on the injected dose or plasma samples were introduced, particularly in case of a reduced clearance. The comparison between the SBS clearances calculated using the 120 min sample to that obtained using the 240 min samples is slightly more sensitive. However, this approach is also slightly less specific. A difference of more than 10 ml . min(-1). 1.73 m(-2) can be observed in the absence of an error. CONCLUSIONS: The use of the SBS for checking the consistency of the 2BS constitutes an insensitive approach to detect an eventual error in the injected dose or in the plasma samples. Obvious different results obtained by SBS and 2BS or between the SBS calculated using the 120 min and the 240 min samples suggest the presence of an error, but comparable results do not exclude erroneous measurement. Moreover, a difference of more than 10 ml . min(-1). 1.73 m(-2) can be observed in the absence of an error in the injected dose or in the plasma samples.

Assessment of lymphocyte-mediated cytotoxicity using flow cytometry.

Cytotoxic lymphocytes, including cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells, kill target cells by releasing granules containing perforin and granzymes, and/or via Fas-Fas ligand interactions. Both pathways lead to prompt activation within target cells of caspase cascades responsible for apoptosis induction and cell death. We have utilized cell-permeable fluorogenic caspase substrates and multiparameter flow cytometry to detect caspase activation in target cells, and applied these tools to quantify and visualize cytotoxic lymphocyte activities. This novel assay, referred to as the flow cytometric cytotoxicity (FCC) assay, is a nonradioactive single-cell-based assay that provides a more rapid, biologically informative, and sensitive approach to measure cytotoxic lymphocyte activity when compared to other assays such as the 51chromium (51Cr) release assay. In addition, the FCC assay can be used to study CTL-mediated killing of primary target cells of different cell lineages that are frequently not amenable to study by the 51Cr release assay. Furthermore, the FCC assay enables evaluation of the phenotype and fate of both target and effector cells, and as such, provides a useful new approach to illuminate the biology of cytotoxic lymphocytes.

Adenosine acts through an A3 receptor to prevent the induction of murine anti-CD3-activated killer T cells.

Adenosine, a purine nucleoside found at high levels in solid tumors, is able to suppress the recognition/adhesion and effector phases of killer lymphocyte-mediated tumor cell destruction. Here, we demonstrate that adenosine, at concentrations that are typically present in the extracellular fluid of solid tumors, exerts a profound inhibitory effect on the induction of mouse cytotoxic T cells, without substantially affecting T-cell viability. T-cell proliferation in response to mitogenic anti-CD3 antibody was impaired in the presence of 10 microM adenosine (plus coformycin to inhibit endogenous adenosine deaminase). Antigen-specific T-cell proliferation was similarly inhibited by adenosine. Anti-CD3-activated killer T (AK-T) cells induced in the presence of adenosine exhibited reduced major histocompatibility complex-unrestricted cytotoxicity against P815 mastocytoma cells in JAM and (51)Cr-release assays. Diminished tumoricidal activity correlated with reduced expression of mRNAs coding for granzyme B, perforin, Fas ligand and tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL), as well as with diminished Nalpha-CBZ-L-lysine thiobenzylester (BLT) esterase activity. Interleukin-2 and interferon-gamma synthesis by AK-T cells was also inhibited by adenosine. AK-T cells express mRNA coding for A(2A), A(2B) and A(3) receptors, but little or no mRNA coding for A(1) receptors. The inhibitory effect of adenosine on AK-T cell proliferation was blocked by an A(3) receptor antagonist (MRS1191) but not by an A(2) receptor antagonist (3,7-dimethyl-1-propargylxanthine [DMPX]). The A(3) receptor agonists (N(6)-2-(4-aminophenyl)ethyladenosine [APNEA] and N(6)-benzyl-5'-N-ethylcarboxamidoadenosine [N(6)-benzyl-NECA]) also inhibited AK-T cell proliferation. Adenosine, therefore, acts through an A(3) receptor to prevent AK-T cell induction. Tumor-associated adenosine may act through the same mechanism to impair the development of tumor-reactive T cells in cancer patients.

Renal function and histopathologic changes in children after liver transplantation.

Up to 60% reduction of renal function has been reported after orthotopic liver transplantation (OLT) in patients receiving cyclosporine. We prospectively investigated renal function and histopathology in 16 children on triple immunosuppression therapy during 3 years after OLT. Cyclosporine was administered in 3 doses/day to preschool children. The median age at OLT was 5.4 years. Determinations of chromium 51-labeled ethylenediaminetetraacetic acid, p = aminohippuric acid, lithium, and sodium clearances, measurements of serum and urinary electrolytes, and urinary concentration tests were performed. Renal biopsy specimens were taken 18 and 36 months after transplantation. The mean glomerular filtration rate was 121.5 ml/min per 1.73 m2 before transplantation, 86.3 at discharge, and 119.4 36 months after OLT. Hyperuricemia, hyperkalemia, and reduced urinary concentrating capacity were common. Hyperkalemia occurred in 13% to 19% of the patients, only during the first 6 months. Hyperuricemia and reduced concentrating capacity occurred with incidences of 17% to 44% and 40% to 63%, respectively. Histopathologic changes were mild, and severe nephrotoxic effects of cyclosporine was not seen. However, tubular atrophy, mesangial matrix increase, and mesangial cell proliferation were common. We conclude that triple immunosuppression with cyclosporine administration, in three doses per day, to small children and careful renal follow-up ensure good renal function after OLT.