Evaluation of the turnaround time of an integrated preanalytical and analytical automated modular system in a medium-sized laboratory.

OBJECTIVES: Evaluation of an integrated Modular Preanalytics (MPA) and Modular Analytics SWA (MA) system (Roche Diagnostics) during continuous batch processing. DESIGN AND METHODS: A total of 1000 blood specimen tubes was processed and tested in a batch-wise fashion, according to two different specimen input conditions (Study 1 and Study 2). The resulting turnaround time of the system was assessed. RESULTS: Study 1 tubes were centrifuged in the MPA. The preanalytical time rose steadily from 14 to 28 min, and after sample 315, it showed minimal variation. The analytical time remained almost constant. In Study 2, tubes were centrifuged before being processed in the MPA. The preanalytical time increased from 4 to 19 min, and the analytical time increased similarly. The turnaround time in Study 1 was 132 min and in Study 2 was 108 min. CONCLUSION: Centrifugation in the MPA slightly increased the turnaround time. Nevertheless, the labor associated with specimen processing was reduced and the efficiency of the laboratory was improved.

Copper metabolism and biliary secretion in patients receiving orthotopic liver transplantation.

BACKGROUND: The quantitative aspects of biliary copper excretion in health and disease have not been fully defined yet. The aim of the study was to evaluate copper metabolism and biliary excretion of patients who have received an orthotopic liver transplant (OLT) during the immediate postoperative period. METHODS: We have studied retrospectively 16 patients undergoing primary OLT and eight undergoing cholecystectomy, and measured serum concentration of copper and its secretion in bile and urine by flame atomic absorption spectrometry (FAAS). RESULTS: We found a progressive increase of biliary copper secretion rates and a corresponding lowering of urinary copper during the postoperative period. Thus, in OLT patients, the mean of biliary copper secretion on day 1 is 0.7+/-0.2 micromol/day compared with 2.3+/-1.1 micromol/day on day 7 (p<0.01) and 6.1+/-2.5 micromol/day on day 15 (p<0.0001). The rate of copper output on day 5 after surgery is about one sixth of the value reported for patients who had undergone cholecystectomy. In patients suffering an acute rejection episode, there was an abrupt fall in bile flow (<15 ml/day) and excretion of biliary copper (<1 micromol/day), accompanied by an increase of urine copper excretion (>3 micromol/day), and both were recovered when the rejection episode was solved. We found an inverse relationship between the serum bilirubin (Bt), alkaline phosphatase (ALP) and the biliary copper excretion (p<0.01), and a direct relationship with urinary copper excretion (p<0.01). CONCLUSIONS: The copper measurements in urine and bile are non-invasive techniques, of low cost, rapid and easy to accomplish, and available in hospitals accredited for hepatic transplantation. These characteristics make these methods helpful in the monitoring of patients submitted to OLT for assessment of graft quality and subsequent outcome.

Serum ionized magnesium monitoring during orthotopic liver transplantation.

During orthotopic liver transplantation (OLT) citrate accumulates and magnesium can be chelated, which can lead to ionized hypomagnesemia and cardiovascular dysfunction. Our aim was to study the serum ionized magnesium (Me2+) evolution and establish its relation to serum total Mg and citrate levels during OLT. We studied 58 adult patients undergoing OLT. The serum Me2+ level dropped significantly at the end of the preanhepatic phase, and remained low until the end of the procedure. Furthermore, the Me2+ levels remained below the range of reference from the beginning of the anhepatic phase onward. There was an inverse correlation between Me2+ and citrate for all patients. Me2+, like ionized calcium (Ca2+), is chelated by citrate and its evolution is a mirror image of that of citrate. In our patients, we did not observe any significant dysrhythmias that could be directly attributed to ionized hypomagnesemia. In conclusion, low preoperative levels, together with the massive transfusion of blood products and the increase in renal losses, cause progressive ionized hypomagnesemia in OLT patients. We propose that it he routinely monitored and treated accordingly, as is already done with Ca2+.

The value of polymorphonuclear elastase in adult respiratory distress syndrome.

The clinical usefulness of quantitative plasma polymorphonuclear elastase (PMN-elastase) determinations as prognostic markers of adult respiratory distress syndrome (ARDS) in polytraumatized patients was analyzed. PMN-elastase and C-reactive protein (CRP) levels were determined in 55 polytraumatized patients admitted into the Intensive Care Unit. Eight patients developed ARDS and 47 patients did not. These parameters were also analyzed in a control group (n = 34). PMN-elastase levels in ARDS cases reached significantly higher values than in patients who did not develop this syndrome (P < 0.01). We conclude that the increase in plasma PMN-elastase levels can be useful in predicting the development of ARDS in polytraumatized patients, in instituting prophylactic actions and monitoring the course of the disease in these high risk patients. This test is easily adaptable to the routine of any hospital laboratory.

Amphiphilic and hydrophilic forms of acetyl- and butyrylcholinesterase in human brain.

Human brain acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) were sequentially extracted, first with a Tris-saline buffer (S1) and then with 1% (w/v) Triton X-100 (S2). About 20 and 30% of the AChE and BuChE activities were recovered in S1 and most of the remaining enzymes in S2. Main molecular forms of about 10.5 S and 12.0 S, G4 forms of AChE and BuChE, and smaller amounts of 4.5 S and 5.5 S forms, G1 species of AChE and BuChE, were measured in S1. Application of Triton X-114 phase partitioning and affinity chromatography on phenyl-agarose to S1 revealed that 25% of the AChE and none of the BuChE molecules displayed amphiphilic properties. Analysis of the enzyme activity retained by the phenyl-agarose showed that G1 AChE constituted the bulk of the amphiphilic molecules released without detergent. Main G4 forms of AChE and BuChE were found in the S2 extract. Eighty and 45% of the AChE and BuChE activities in S2 were measured in the detergent-rich phase by Triton X-114 phase partitioning. Thus, most of the AChE and about half of the BuChE molecules in S2 displayed amphiphilic properties. The main peak of BuChE, a 12.0 S form in gradients made with Triton X-100, splits into two peaks of 9.5 S and 12.5 S in Brij 96-containing gradients. This suggests that hydrophilic G4 BuChE forms are predominant in S1 and that hydrophilic and amphiphilic isoforms coexist in S2.

Ricinus communis agglutinin I reacting and non-reacting butyrylcholinesterase in human cerebrospinal fluid.

Differences in glycosylation of acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) in human brain, plasma and cerebrospinal fluid (CSF) have been investigated by means of their interaction with agarose-immobilized lectins. Most of the AChE in brain and CSF was associated to concanavalin A (Con A), Lens culinaris (LCA) and Triticum vulgaris (WGA) agglutinins, but little activity was adsorbed to Ricinus communis agglutinin I (RCAI). Brain, plasma and CSF BuChE was almost fully bound to Con A, LCA and WGA-agarose. Brain BuChE was unable to react with RCA (RCA-BuChE), the plasma enzyme was completely bound to the lectin (RCA+BuChE) and BuChE from CSF of normal children was partially fixed to RCA (RCA +/- BuChE). BuChE in CSF of children with meningitis fully reacts with the lectin. The data suggest that the proportion of RCA+BuChE in CSF of children with meningitis is increased, this enzyme probably coming from plasma.