Role of Technology for the Management of AKI in Critically Ill Patients: From Adoptive Technology to Precision Continuous Renal Replacement Therapy.

This paper reports on the continuous renal replacement therapy (CRRT) technology group recommendations and research proposals developed during the 17th Acute Dialysis Quality Initiative Meeting in Asiago, Italy. The group was tasked to address questions related to the impact of technology on acute kidney injury management. We discuss technological aspects of the decision to initiate CRRT and the components of the treatment prescription and delivery, the integration of information technology (IT) on overall patient management, the incorporation of CRRT into other 'non-renal' extracorporeal technologies such as ECMO and ECCO2R and the use of sorbents in sepsis and propose new areas for future research. Instead of reviewing current knowledge, the group focused on developing a renovated research agenda that reflects current and future technological advances, centered on innovations in new equipment, membranes and IT that will permit the integration of patient care and decision-making processes for years to come.

Phenotypic, genetic and genomic consequences of natural and synthetic polyploidization of Nicotiana attenuata and Nicotiana obtusifolia.

BACKGROUND AND METHODS: Polyploidy results in genetic turmoil, much of which is associated with new phenotypes that result in speciation. Five independent lines of synthetic allotetraploid N. x obtusiata (N x o) were created from crosses between the diploid N. attenuata (Na) (male) and N. obtusifolia (No) (female) and the autotetraploids of Na (NaT) and No (NoT) were synthesized. Their genetic, genomic and phenotypic changes were then compared with those of the parental diploid species (Na and No) as well as to the natural allotetraploids, N. quadrivalvis (Nq) and N. clevelandii (Nc), which formed 1 million years ago from crosses between ancient Na and No. KEY RESULTS: DNA fingerprinting profiles (by UP-PCR) revealed that the five N x o lines shared similar but not identical profiles. Both synthetic and natural polyploidy showed a dosage effect on genome size (as measured in seeds); however, only Nq was associated with a genome upsizing. Phenotypic analysis revealed that at the cellular level, N x o lines had phenotypes intermediate of the parental phenotypes. Both allo- and autotetraploidization had a dosage effect on seed and dry biomass (except for NaT), but not on stalk height at first flower. Nc showed paternal (Na) cellular phenotypes but inherited maternal (No) biomass and seed mass, whereas Nq showed maternal (No) cellular phenotypes but inherited paternal (Na) biomass and seed mass patterns. Principal component analysis grouped Nq with N x o lines, due to similar seed mass, stalk height and genome size. These traits separated Nc, No and Na from Nq and N x o lines, whereas biomass distinguished Na from N x o and Nq lines, and NaT clustered closer to Nq and N x o lines than to Na. CONCLUSIONS: Both allo- and autotetraploidy induce considerable morphological, genetic and genomic changes, many of which are retained by at least one of the natural polyploids. It is proposed that both natural and synthetic polyploids are well suited for studying the evolution of adaptive responses.

Resistance of vascular access catheters for continuous renal replacement therapy: An ex vivo evaluation.

AIM: To assess the resistance posed by double-lumen vascular access dialysis catheters at low and high blood flow. DESIGN: Controlled ex vivo study Setting: ICU Laboratory of tertiary hospital. SUBJECTS: Eleven proprietary vascular access catheters for continuous renal replacement therapy. METHODS: Heparinized spent red cells diluted in polygeline solution were pumped using the Aquarius hemofiltration machine (Edwards Life Sciences, Sydney, NSW, Australia) and its standard circuit through several vascular access catheters. Blood flow was increased and then decreased in steps of 50 ml/min (50, 150, 200, 250 and 300 ml/min) while catheter outflow and inflow pressures were recorded. The pressure-flow relationship (hydraulic resistance) of each catheter was then calculated. Study catheters were divided into two groups according to their internal diameter (large gauge vs. smaller gauge) or length (long or short). Hydraulic resistances were compared between the groups. RESULTS: Different double lumen catheters posed clearly different resistances to flow. For all groups of catheters, there was a linear relationship between pressure and flow. No statistically significant difference between short and long catheters could be demonstrated (p=0.715). On the other hand, larger gauge catheters (13 Fr or greater) had significantly lower resistances than smaller gauge (<13 Fr) catheters (p=0.0062). Furthermore, all larger gauge catheters had resistances lower than 0.430 mmHg/ml/min, while all smaller gauge catheters had resistances greater than 0.490 mmHg/ml/min. CONCLUSIONS: Commercial double-lumen dialysis catheters have variable resistance to blood flow under standard ex vivo conditions. Although both length and internal diameter varied, internal diameter had a dominant effect on resistance. This information might be useful to clinicians in guiding their choice of catheters for clinical use.

The effect of three different miniaturized blood purification devices on plasma cytokine concentration in an ex vivo model of endotoxinemia.

PURPOSE: A novel type of adsorptive plasma filtering device (ETX-A) capable of removing endotoxin from blood in a single step has recently been developed using nanotechnology. METHODS: In a miniaturized, ex vivo model of extracorporeal circuits, we tested the capacity to reduce plasma cytokine concentration of ETX-A filters in comparison to standard high-flux (HF) filters, high cut-off (HCO) filters and a control. Blood from six healthy volunteers was spiked with endotoxin and then circulated through closed (ETX-A, control) or open (HF, HCO) circuits. Blood flow was set at 16 ml/min and filtration flow at 1 ml/min. Samples for measurement of IL-1ra and IL-6 were taken at baseline and at 4 hours. RESULTS: Compared to control (703.3 [850.6] pg/mL), in HCO (383.5 [1144.1] pg/mL) and ETX-A (490.1 [683.2] pg/mL) filters, plasma IL-1ra pooled pre- and postfilter concentrations were lower at the end of the experiment (P=0.002; P=0.050, respectively) whereas, in standard HF filters, IL-1ra concentration was higher than control. HCO showed a trend toward a reduced relative increase in IL-6 concentration from commencement to end of experiment compared to control (P=0.07). After pooling end-of-experiment plasma cytokine values of novel blood purification devices, we found HCO + ETX-A superior to H with regard to reduction of IL-1ra (-27.0 [-20.5]% vs. 8.1 [18.9]%; p<0.001) and IL-6 (-18.0 [38.3]% vs. -1.1 [24.3]%; P=0.050) compared to control. CONCLUSIONS: HCO and ETX-A appeared to significantly reduce plasma IL-1ra and, when combined, plasma IL-6 concentration as well. It appears desirable to manufacture full-size blood purification devices using this technology and to explore their effect on cytokine removal.

The acid-base effect of changing citrate solution for regional anticoagulation during continuous veno-venous hemofiltration.

PURPOSE: To compare the acid-base balance effects of two different citrate doses for regional citrate anticoagulant (RCA) for continuous veno-venous hemofiltration (CVVH). METHODS: We used a commercial citrate fluid (citrate concentration: 11 mmol/L) from July 2003 to July 2004 (period A) in 22 patients; then changed to a new citrate fluid (citrate concentration: 14 mmol/L) from July 2004 to Feb 2005 (Period B) in 21 patients. Replacement fluid rate was fixed at 2,000 ml/h. We measured all relevant variables for acid-base analysis according to the Stewart-Figge methodology. RESULTS: After commencement of RCA-CVVH, there was a change in bicarbonate and base excess (BE) toward acidosis for both fluids. This change was significantly different between period A and B at 6 and 12 hours (pH: p<0.01, BE: p<0.05) with greater decreases with the 11 mmol/L citrate fluid. These changes were mostly secondary to an increase in the strong ion difference (SID) and occurred despite an increased strong ion gap (SIG) (+0.5 mEq/L vs. +1.5 mEq/L; p<0.01) in the higher citrate concentration fluid. Cessation of RCA-CVVH was associated with short-lived differences in bicarbonate and SIG which were similar to those seen on initiation of RCA-CVVH but in the opposite direction. CONCLUSIONS: A small increase This was partly offset by an increase in SIG, consistent with increased citratemia. Cessation of treatment showed a differential improvement in SIG also consistent with disposal of therapy-associated citrate. These observations might assist clinicians in interpreting acidbase changes during RCA-CVVH.in citrate infusion rate caused an alkalinizing increase in SID.

Beta2-microglobulin removal and plasma albumin levels with high cut-off hemodialysis.

PURPOSE: beta2-microglobulin (beta2MG) is pivotal to the pathogenesis of dialysis-related amyloidosis. We compared the effects of high cut-off hemodialysis (HCO-HD) with those of standard high-flux hemodialysis (HF-HD) regarding the concentration and clearance of beta2MG and albumin. DESIGN: We enrolled ten patients with acute renal failure in a double-blind, cross-over, randomized controlled trial. PROCEDURES: Each patient received four hours of HCO-HD (estimated in vivo cutoff 50-60 kDa) and four hours of HF-HD (estimated in vivo cutoff 15-20 kDa) in random order. Statistical methods and outcome measures: As data lacked normal distribution, we used nonparametric statistical analysis. Plasma and dialysate concentrations of beta2MG and albumin were measured at baseline and after four hours of each study treatment. MAIN FINDINGS: We found significantly greater diffusive beta2MG clearances for HCO-HD compared to HF-HD (at the start: 71.8 ml/min vs. 5.1 ml/min; P=0.008 and at the end: 68.8 ml/min vs. 5.7 ml/min; P=0.008). We found a reduction in plasma beta2MG concentrations of -31.6% during HCO-HD compared to an increase by 25.7% during HF-HD; P=0.008. At baseline (HCO-HD: 26.0 g/L vs. HF-HD: 26.5 g/L), and at the end of both treatments, plasma albumin concentrations were comparable (HCO-HD: 25.5 g/L vs. HF-HD: 26.5 g/L; P=0.25). During HCO-HD, albumin clearance was 1.9 ml/min at the start and decreased significantly to 0.8 ml/min at the end; P=0.008. HF-HD had an albumin clearance of 0.01 ml/min. CONCLUSIONS: HCO-HD was more effective in decreasing plasma beta2MG concentrations than standard HF-HD and did not reduce plasma albumin levels. Further studies of HCO-HD in the treatment of dialysis-related beta2MG accumulation appear warranted.

Fluid balance error in continuous renal replacement therapy: a technical note.

BACKGROUND: The reliability and safety of continuous renal replacement therapy (CRRT) machines have improved, yet there still remains the potential for fluid balance errors to occur during treatment. METHODS: In vitro testing of two Kimal Hygieia CRRT machines (Plus and Ultima) was performed. Normal saline to simulate the blood circuit and standard bicarbonate-based fluid for replacement were used. All tests were performed in CVVH mode at four ultrafiltration (UF) rates. The testing was based on creation of a voluntary fluid balance error by clamping the line that fills the replacement fluid chamber to stop flow to the (simulated) patient. The time to alarms and fluid balance errors were recorded. The alarms were overridden and the accumulated fluid balance error allowed by the machine was determined. RESULTS: The alarm occurred approximately 1 minute after the replacement fluid line was clamped at all UF rates. There was no limit to the number of times the alarm could be overridden and the accumulated negative fluid balance was proportional to the prescribed UF rate. After the replacement fluid chamber was allowed to re-fill, the machine attempted to correct the fluid deficit and consistently delivered excess fluid to generate a positive fluid balance error. CONCLUSIONS: The Hygieia machines appear designed with appropriate alarm and safety features. However, simulated fluid balance errors raise caution for operators. Clinicians and nurses need to understand the clinical implications of alarm overrides. Fluid balance errors caused by failure to acknowledge and correct replacement fluid failure alarms may cause harm to patients.

A pilot randomized controlled crossover study comparing regional heparinization to regional citrate anticoagulation for continuous venovenous hemofiltration.

OBJECTIVE: To evaluate the efficacy and safety of a regional heparinization and a regional citrate method of anticoagulation in CVVH. DESIGN: Randomized controlled cross-over study. SUBJECTS: Ten critically ill patients with acute renal failure. SETTING: ICU of tertiary hospital. INTERVENTION: CVVH was performed with pre-filter fluid replacement at 2000 ml/h and a blood flow rate of 150 ml/min. Regional heparinization was by the administration of heparin pre-filter at 1500 IU/h and protamine post-filter at 15 mg/h. Regional citrate anticoagulation was by means of a citrate-based replacement fluid (14 mmol/L) administered pre-dilution. RESULTS: We studied nine males and one female. The mean age and APACHE II score were 70.5 and 17 respectively. Median circuit life was 13 hours (IQR 9.28) for the regional heparinization method compared to 17 hours (IQR 12,19.5) for the regional citrate method (p=0.77). There were no episodes of bleeding in either group. CONCLUSION: Regional heparinization and regional citrate anticoagulation achieve similar circuit life in critically ill patients receiving CVVH.

High cut-off point membranes in septic acute renal failure: a systematic review.

OBJECTIVES: To review the literature on the experimental, physiological and clinical effects of blood purification with high cut-off (HCO) point membranes in septic acute renal failure (ARF). STUDY DESIGN: MEDLINE and PubMed database search combining relevant terms and integrating data from studies on the use of HCO membranes. SETTING AND POPULATION: Ex vivo studies of endotoxemia, animal studies of bacteremia and clinical studies using HCO membranes in patients with septic ARF. SELECTION CRITERIA FOR STUDIES: Original data from primary publications. INTERVENTIONS: HCO membrane-based hemodialysis, hemodiafiltration or hemofiltration. OUTCOMES: Plasma cytokine clearance, immunological and physiological effects and safety parameters of HCO membranes. RESULTS: HCO membranes effectively remove cytokines from blood. Treatment using HCO membranes has beneficial effects on immune cell function and increases survival in animal models of sepsis. Preliminary clinical studies show that HCO membranes decrease plasma cytokine levels and the need for vasopressor therapy. HCO membrane-based blood purification has now been applied in four pilot randomized controlled studies of 70 patients with septic ARF with no reports of serious adverse effects. LIMITATIONS: Because of substantial heterogeneity, no formal quantitative analysis could be performed. CONCLUSIONS: The available evidence on HCO blood purification justifies larger randomized controlled trials in patients with septic ARF.

A pilot randomized controlled comparison of extended daily dialysis with filtration and continuous veno-venous hemofiltration: fluid removal and hemodynamics.

OBJECTIVES: Extended intermittent dialytic techniques are increasingly being reported in the treatment of ARF in the ICU but few randomized controlled trials exist. We compared one such technique to a technique of continuous renal replacement therapy with regard to fluid removal and hemodynamics. METHODS: Sixteen critically ill patients with ARF were enrolled in a randomized controlled trial at the ICU of a tertiary hospital. We randomized eight patients to three consecutive days of treatment with either Extended Daily Dialysis with filtration (EDDf) or Continuous Veno-Venous Hemofiltration (CVVH) and compared fluid removal and hemodynamics during treatment. RESULTS: A total of 16.6 liters of fluid were removed during EDDf (830 mL/day over 20 treatment days) compared with 15.4 liters (700 ml/day over 22 treatment days) during CVVH. Median fluid removal per day was 1837 mL in the EDDf group compared with 1410 mL per day in the CVVH group, p=0.674. Median hourly fluid removal rate was 252 mL for EDDf and 128 mL for CVVH (p<0.01). Mean arterial pressure in the EDDf group was lower at two hours after starting treatment (76 mmHg vs. 94 mmHg) in the CVVH group; p= 0.031. There was no significant difference between groups for heart rate, CVP and noradrenaline dose at all time intervals measured. CONCLUSIONS: Adequate prescribed fluid removal was achieved with both techniques. However, as expected, fluid was removed at a faster rate during EDDf. This was initially associated with a lower blood pressure than during CVVH where blood pressure increased.

Nurses' attitudes to a medical emergency team service in a teaching hospital.

BACKGROUND: Cultural barriers including allegiance to traditional models of ward care and fear of criticism may restrict use of a medical emergency team (MET) service, particularly by nursing staff. A 1-year preparation and education programme was undertaken before implementing the MET at the Austin Hospital, Melbourne, Australia. During the 4 years after introduction of the MET, the programme has continued to inform staff of the benefits of the MET and to overcome barriers restricting its use. OBJECTIVE: To assess whether nurses value the MET service and to determine whether barriers to calling the MET exist in a 400-bed teaching hospital. METHODS: Immediately before hand-over of ward nursing, we conducted a modified personal interview, using a 17-item Likert agreement scale questionnaire. RESULTS: We created a sample of 351 ward nurses and obtained a 100% response rate. This represents 50.9% of the 689 ward nurses employed at the hospital. Most nurses felt that the MET prevented cardiac arrests (91%) and helped manage unwell patients (97%). Few nurses suggested that they restricted MET calls because they feared criticism of their patient care (2%) or criticism that the patient was not sufficiently unwell to need a MET call (10%). 19% of the respondents indicated that MET calls are required because medical management by the doctors has been inadequate; many ascribed this to junior doctors and a lack of knowledge and experience. Despite hospital MET protocol, 72% of nurses suggested that they would call the covering doctor before the MET for a sick ward patient. However, 81% indicated that they would activate the MET if they were unable to contact the covering doctor. In line with hospital MET protocol, 56% suggested that they would make a MET call for a patient they were worried about even if the patient's vital signs were normal. Further, 62% indicated that they would call the MET for a patient who fulfilled MET physiological criteria but did not look unwell. CONCLUSIONS: Nurses in the Austin Hospital value the MET service and appreciate its potential benefits. The major barrier to calling the MET appears to be allegiance to the traditional approach of initially calling parent medical unit doctors, rather than fear of criticism for calling the MET service. A further barrier seems to be underestimation of the clinical significance of the physiological perturbations associated with the presence of MET call criteria.

Management of fluid balance in CRRT: a technical approach.

BACKGROUND: The possibility of making fluid balance errors during continuous renal replacement therapy has been identified since the beginning of this modality of treatment. The advent of automated machines has partially overcome this problem. Nevertheless, there are conditions and operation modes in which the potential for fluid balance errors is still present. OBJECTIVE: To analyse fluid balance management in CRRT therapies across a range of currently marketed machine. METHODS: The tests were conducted in vitro, utilizing saline solution for the blood circuit and regular dialysate/reinfusate for the dialysate/reinfusion circuit. The methodology used was based on the voluntary creation of a fluid balance error by altering the correct flow in the circuit of the different machines. Subsequently, the time for alarm occurrence and the threshold value for fluid balance error was evaluated. The alarm was overridden and the overall fluid error allowed by the machine was evaluated. Each machine was tested in conditions of different dialysate/filtrate flow rates and in different simulated treatment modalities. RESULTS: Fluid balance errors can be easily avoided not only by a correct and careful adherence to the protocols of use of the current CRRT machines, but also by the compliance to prescriptions and programmed controls during therapy. Most importantly, if an alarm appears on the machine, one can try to override it without major problems; major problems may occur when multiple override commands are operated without identifying the problem and solving it adequately. CONCLUSION: Machines seem to be designed with adequate safety features and accurate alarm systems. However, features and alarms can be manipulated by operators creating the opportunity for serious error. Physicians and nurses involved in prescription and delivery of CRRT should have precise protocols and defined procedures in relation to machine alarms to prevent major clinical problems.

Commercial low-citrate anticoagulation haemofiltration in high risk patients with frequent filter clotting.

This study assessed the safety and efficacy of a commercial low-citrate concentration-based pre-filter replacement fluid during continuous veno-venous haemofiltration (CVVH) in patients with frequent filter clotting and high risk of bleeding. We used a commercial low-citrate fluid as pre-dilution replacement fluid during CVVH (citrate: 11 mmol/l (33 meq/l), sodium: 140 mmol/l, chloride: 108 mmol/l and potassium: 1 mmol/l). A calcium and magnesium infusion was delivered separately by central line for the maintenance of serum ionized calcium (Cai) and total magnesium (Mg). In this prospective observational study, 30 patients, 124 filters and 1,515 treatment-hours were observed. Median filter life of citrate CVVH was 9.5 hours. Filter life in the 48 hours prior to citrate CVVH was also observed. In the patients on prior non-anticoagulant CVVH (n=14) filter life increased significantly with citrate (9.5 hours vs 5 hours; P<0.0001). In patients on prior heparin CVVH (n = 15), filter life was similar with citrate (10 hours vs 8 hours; P = 0.68). However, in patients with prior early/frequent filter clotting despite heparin (n = 11) filter life increased significantly (10 hours vs 7 hours; P=0.038). Of 411 serum Cai measurements, none showed a Cai < 0.85 mmol/l and, of 84 observations, none showed a serum Mg<0. 6 mmol/l. One patient with sepsis and shock needed to cease citrate CVVH because of progressive ionized hypocalcaemia and increasing anion gap. No other adverse effects were observed. In selected patients, CVVH with a commercial low-citrate concentration solution as pre-filter replacement fluid and a simultaneous calcium and magnesium infusion protocol appears generally safe. Filter life was acceptable and superior to that achieved with previous treatment.

[Long-term intermittent renal replacement therapy at an intensive care unit].

Standard intermittent hemodialysis (IHD) used for the treatment of acute renal failure (ARF) at an intensive care unit has significant biochemical and physiological drawbacks. In the past 20 years, these drawbacks have stimulated the development of continuous renal replacement therapy (CRRT) and its ever-increasing use. However, CRRT is technically complicated and requires 24-hour monitoring. In some clinics, the use of CRRT leads to that each patient is under his/her nurse's surveillance, instead 1 nurse per 2 patients as before; this change has economic consequences and may limit nursing accessibility to other patients. The procedures prolonging intermittent therapy do not require 24-hour monitoring may benefit the treatment of ARF at the intensive care therapy. In this paper the authors call such procedures for continuous intermittent renal replacement therapy. They are characterized by a number of basic principles: (1) the use of modified or standard dialysis apparatuses; (2) the application of diffuse, convection, or both; (3) a certain reduction in the rate of elimination of dissolved substances as compared with IHD; (4) more prolonged treatment: above usual 3 or 4 hours of IHD, but not more than 8-12 hours (hence the term "intermittent"); (5) the use of on-line generation dialysate or substituting fluid. Information on the effectiveness and safety of this procedure is being now compiled.

Low-dose citrate continuous veno-venous hemofiltration (CVVH) and acid-base balance.

OBJECTIVE: To evaluate the acid-base effect of low-dose regional citrate anticoagulation (RCA) during continuous veno-venous hemofiltration (CVVH). DESIGN: Prospective observational study. SETTING: ICUs of tertiary public and private hospitals. SUBJECTS: Thirty critically ill patients with acute renal failure at risk of bleeding or with a major contraindication to heparin-CVVH and/or short filter life. METHODS: We used a commercial citrate-based fluid (11 mmol/L, sodium: 140 mmol/L, chloride: 108 mmol/L and 1 mol/L of potassium) as pre-dilution replacement fluid during CVVH. Further potassium was added according to serum potassium levels. We measured all relevant variables for acid-base analysis according to the Stewart-Figge methodology. RESULTS: Before treatment, study patients had a slight metabolic acidosis, which worsened over 6 hours of RCA-CVVH (pH from 7.39 to 7.38, p < 0.005; bicarbonate from 23.2 to 21.6 mmol/L, p < 0.0001 and base excess from -2.0 to -3.0 mEq/L, p < 0.0001) due to a significant increase in SIG (from 5.8 to 6.6 mEq/L, p < 0.05) and a decrease in SIDa (from 37.5 to 36.6 mEq/L, p < 0.05). These acidifying effects were attenuated by hypoalbuminemia and a decrease in lactate (from 1.48 to 1.34 mmol/L, p < 0.005) and did not lead to progressive acidosis. On cessation of treatment, this acidifying effect rapidly self-corrected within six hours. CONCLUSIONS: Low dose RCA-CVVH induces a mild acidosis secondary to an increased strong ion gap and decreased SIDa which fully self-corrects at cessation of therapy. Clinicians need to be aware of these effects to correctly interpret changes in acid-base status in such patients.

A comparison of two citrate anticoagulation regimens for continuous veno-venous hemofiltration.

AIMS: To assess the safety and efficacy of two different commercial citrate containing pre-filter replacement fluids during continuous veno-venous hemofiltration (CVVH) in patients with frequent filter clotting. SETTING: Four intensive care units. PATIENTS: Sixty-three critically ill patients with acute renal failure (ARF). DESIGN: Prospective observational study. METHODS: We used a commercial citrate fluid (citrate: 11 mmol/L -fluid A) as predilution replacement for CVVH. We then changed to a new commercial citrate fluid (citrate: 14 mmol/L-fluid B) as replacement fluid and performed statistical comparisons. Replacement fluid rate was fixed at 2,000 ml/hour. RESULTS: Filter life was 12.2 hour with fluid A compared with 17.1 hour with fluid B on average (p=0.0001). Mean post filter ionized calcium concentration was 0.52 mmol/L with fluid A compared with 0.40 mmol/L with fluid B (p<0.0001). Citrate intolerance led to cessation of treatment in one patient with fluid A and one patient with fluid B. Overall ionized calcium levels were higher (A: 1.18 vs B: 1.13 mmol/L; p<0.0001) and bicarbonate was lower (A: 22.4 vs B: 24.5 mmol/L; p<0.0001) during treatment with fluid A. Alkalemia was seen in 10 patients treated with fluid A and 16 patients treated with fluid B (NS). CONCLUSIONS: We have developed a simple approach to regional citrate anticoagulation for CVVH using a commercial citrate-containing fluid as replacement fluid. Increasing citrate concentration from 11 to 14 mmol/L increased filter life while maintaining relative safety and simplicity.

An analysis of plant-aphid interactions by different microarray hybridization strategies.

Aphids have long been considered 'stealthy' herbivores that subvert a plant's induced defenses and manipulate its source-sink signaling, but these hypotheses are largely untested at a transcriptional level. We analysed gene expression in native tobacco plants (Nicotiana attenuata) infested with Myzus nicotianae aphids, without resorting to the use of clip-cages, with a cDNA microarray containing 240 defense-related N. attenuata genes. Using a hybridization scheme ('ratio analysis' and 'state analysis') broadly applicable in two-factor analyses, we examined how the aphids influenced source--sink relationships and determined if their feeding preference, apart from benefiting from the sink strength of young leaves, was associated with the expression of known plant defense genes. In contrast to the responses elicited by attack from tissue-feeding lepidopteran larvae and mesophyll-sucking insects, attack from phloem-feeding aphids elicited only weak responses. Similar to other herbivores, M. nicotianae feeding increased the expression of trypsin protease inhibitors (TPI), lipoxygenase, and xyloglucan-endotransglycosylase genes, and decreased small RUBISCO subunit and ubiquitin carrier protein transcripts. Aphid-specific changes included the up-regulation of glutamate synthase and the down-regulation of a germin-like protein. Aphids preferentially settled on younger leaves, which expressed more hydroperoxide lyase and TPI than did older leaves, suggesting that these genes, which mediate the synthesis of compounds reported to be toxic for aphids in other plant systems, are either not under transcriptional control or not important in this system. By identifying aphid-responsive genes, we have made a first step in identifying the 'genes that matter' in plant--aphid interactions.

Prolonged daily intermittent renal replacement therapy in ICU patients by ICU nurses and ICU physicians.

OBJECTIVES: Prolonged daily intermittent renal replacement therapy (PDIRRT) has been proposed as a new form of treatment for severe acute renal failure (ARF). However, this treatment has so far implied a) full dependence on nephrological input, b) lack of any convective clearance and c) limited purification of dialysate water. The aim of this study was to establish the feasibility and safety of performing PDIRRT in the ICU with a) no nephrological input, b) the addition of some convective clearance with on-line fluid replacement and c) a new advanced water purification system. DESIGN: Prospective observational study. PATIENTS: Fourteen patients treated with PDIRRT. SETTING: ICU of tertiary institution. INTERVENTIONS: Treatment of patients with severe ARF and critical illness with PDIRRT. Prescription of treatment by ICU physicians. Conduct of treatment by ICU nurses. Use of combined convective and diffusive therapy with on-line generation of fluid replacement, application of a double-filtration water purification system. MEASUREMENTS AND MAIN RESULTS: We prospectively collected demographic, biochemical, hemodynamic and clinical data in 14 patients, who received 30 PDIRRT treatments for a cumulative treatment time of 205.4 hours. The mean age was 57.9 +/- 16.0. Eight patients were male and 6 female. Their mean APACHE II score was 24.6 +/- 5.9 and their SAPS II score was 41.7 +/- 18.8. PDIRRT was used after at least 24 hours of initial stabilization with continuous veno-venous hemofiltration (CVVH). Blood flow was kept at 100 ml/min dialysate flow at 200 ml/min and convective clearance varied from 21 ml/min to 33 ml/min. All patients were either anuric or oliguric (UO < 400 ml/day). Ten patients were on mechanical ventilation and 11 patients on vasopressor support. Mean treatment session time was 6.9 +/- 1.8 hours. The mean pre-PDIRRT urea was 19.2 +/- 6.9 mmol/L and the creatinine was 274 +/- 116 micromol/L. The mean pre-PDIRRT lactate was 2.95 +/- 2.24 mmol/L. Following treatment, all had significantly decreased to 13.2 +/- 6.3 mmol/L, 215 +/- 95 micromol/L and 2.25 +/- 1.61 mmol/L, respectively (p=<0.0001, <0.0001, <0.05). Bicarbonate levels remained stable during treatment (23.0 +/- 3.8 mmol/L to 23.1 +/- 2.5 mmol/L). Mean norepinephrine dose changed from 8.8 +/- 11.9 microg/min to 12.9 +/- 27.0 microg/min after treatment (NS). There were no complications of therapy. Patient ICU survival was 71.4%. CONCLUSIONS: PDIRRT with combined diffusive and convective clearance is an efficacious form of renal replacement, which can be safely and effectively conducted by ICU nurses following prescription by ICU physicians without any nephrological involvement and with adequate double filtration water purification.

Super high flux hemodialysis at high dialysate flows: an ex vivo assessment.

BACKGROUND AND OBJECTIVES: The removal of cytokines by standard hemofiltration is limited. Super high flux membranes may significantly improve removal even when used in dialysis mode. We sought to measure cytokine clearance using a large surface super high-flux membrane and a standard hemodialysis setting. SETTING: ICU laboratory of a tertiary institution. SUBJECTS: Six healthy volunteers. METHODS: Blood form healthy volunteers was incubated for 4 hours with E. coli endotoxin to stimulate cytokine production. Cytokine containing blood was then circulated through a dialysis circuit at 3 different dialysate flow rates. Blood and dialysate were sampled for cytokine and albumin measurements and calculation of clearances. RESULTS: Super high-flux dialysis achieved high median cytokine clearances (IL-1 clearance of 106 ml/min, IL-6 clearance of 66.8 ml/min, IL-8 clearance of 61.7 ml/min and TNF clearance of 36.1 ml/min). Increasing dialysate flow rate from 300 to 500 ml/min did not significantly increase cytokine clearances. Albumin clearances however were between 2.7 and 5.4 ml/min. CONCLUSIONS: Cytokine dialysis is feasible at high dialysate flow rates yielding high cytokine clearances. Albumin loss, however, is appreciable and may require separate supplementation in the clinical setting.

A pilot study of the effect of altering airway pressure on systolic pulse pressure variation in the systemic and pulmonary arterial circulations.

OBJECTIVE: Systolic pressure variation results from cyclical fluctuation in the intra-thoracic pressure associated with mechanical ventilation and has been used as a measure of relative hypovolemia in mechanically ventilated patients. The impact of the magnitude of the tidal volume and airway pressure on systolic pressure variation, however, has not been examined in mechanically ventilated patients. METHODS: Two patients underwent monitoring following elective cardiac surgery. Tidal volume was randomly varied between 3 and 11 mL/kg over a two minute interval, and the corresponding airway pressure was monitored, as were the effects on the systolic pressure variation of the systemic and pulmonary circulations. RESULTS: There was a strong correlation between increasing tidal volume and peak airway pressure (p<0.0001). In addition, peak airway pressure strongly correlated with the systolic pressure variation of both the systemic and pulmonary circulations (p<0.0001). The increase in diastolic pulmonary arterial pressure induced by insufflation correlated well with the associated increase in systolic blood pressure (p<0.0001). Similarly, the increase in systolic pulmonary artery pressure (PAP) correlated with the associated decrease in systolic blood pressure induced by insufflation (p<0.0001). CONCLUSIONS: Systolic pressure variation in the systemic and pulmonary circulations is affected by tidal volume and peak airway pressure. This should be considered when using systolic pressure variation as a marker of intravascular volume status. Our findings regarding the correlations between changes in the pulmonary arterial pressure and the systemic arterial pressure induced by mechanical ventilation are consistent with the proposed physiological mechanisms of systolic pressure variation.