Targeting inflammation to influence cognitive function following spinal cord injury: a randomized clinical trial.

STUDY DESIGN: This study was a randomized, parallel-group, controlled clinical trial. OBJECTIVES: The purpose of this study was to examine the efficacy of targeting inflammation as a means of improving cognitive function in individuals with spinal cord injury. SETTING: Participants were recruited from the Niagara region of Ontario Canada and all testing occurred on-site at Brock University. METHODS: Indices of memory and verbal learning were assessed by means of the California Verbal Learning Test (CVLT). Inflammation and concentrations of neuroactive compounds related to the kynurenine pathway were assessed via a number of pro- and anti-inflammatory cytokines, as well as tryptophan, kynurenine and several large neutral amino acids. All assessments were performed at baseline as well as at 1 month and 3 months during a 3-month intervention by means of an anti-inflammatory diet. RESULTS: Despite a reduction in inflammation, all measures of the CVLT, including list A, trial 1 (P=0.48), learning slope (P=0.46), long delay free recall (P=0.83), intrusions (P=0.61) and repetitions (P=0.07), showed no significant group × time interaction. CONCLUSION: It may be possible that the reduction in inflammation achieved in the current study was insufficient to induce substantial changes in indices of verbal learning and memory. Alternatively, as these participants likely underwent years of previous chronic inflammation, the underlying hippocampal damage may have negated potential improvements induced by acute reductions in inflammation.

Effectiveness of etoposide chemomobilization in lymphoma patients undergoing auto-SCT.

The effectiveness of stem cell mobilization with G-CSF in lymphoma patients is suboptimal. We reviewed our institutional experience using chemomobilization with etoposide (VP-16; 375 mg/m(2) on days +1 and +2) and G-CSF (5 µg/kg twice daily from day +3 through the final day of collection) in 159 patients with lymphoma. This approach resulted in successful mobilization (>2 × 10(6) CD34+ cells collected) in 94% of patients (83% within 4 apheresis sessions). Fifty-seven percent of patients yielded at least 5 × 10(6) cells in 2 days and were defined as good mobilizers. The regimen was safe with a low rate of rehospitalization. Average costs were $14 923 for good mobilizers and $27 044 for poor mobilizers (P<0.05). Using our data, we performed a 'break-even' analysis that demonstrated that adding two doses of Plerixafor to predicted poor mobilizers at the time of first CD34+ cell count would achieve cost neutrality if the frequency of good mobilizers were to increase by 21%, while the frequency of good mobilizers would need to increase by 25% if three doses of Plerixafor were used. We conclude that chemomobilization with etoposide and G-CSF in patients with lymphoma is effective, with future opportunities for cost-neutral improvement using novel agents.

Decreased motor unit discharge rate in the potentiated human tibialis anterior muscle.

AIM: The purpose of this study was to examine the influence of post-activation potentiation (PAP), the transient increase in low-frequency isometric force observed after muscle activity, on motor unit discharge rates measured during submaximal contractions. METHODS: A quadrifilar needle electrode was inserted into the tibialis anterior muscle to determine discharge rate of individual motor units while monopolar electrodes were used to monitor the root-mean-square (RMS) and mean power frequency (MPF) of the surface EMG signal. Control (unpotentiated) and experimental (potentiated) measures were obtained during a 5 s voluntary contraction at 50% of maximal. In between these measures, subjects performed a 10 s maximal voluntary contraction (MVC) to induce PAP. RESULTS: All subjects data are reported as means ± SEM (n = 10). Compared to baseline values measured prior to the MVC, isometric twitch force measured immediately after the MVC was increased by 260 ± 16% (day 3). Motor unit discharge rate in the potentiated tibialis anterior muscle decreased by approx. 10%, from 20.3 ± 0.8 (before) to 18.3 ± 0.99 pps (P = 0.01) (after). Moreover, the MPF was decreased by approx. 9% (from 58.1 ± 2.84 to 53.6 ± 2.85 Hz; P = 0.01) in the potentiated tibialis anterior. On the other hand, consistent with the absence of fatigue during the MVC, the RMS signal was not altered in the potentiated tibialis anterior (0.29 ± 0.03 vs. 0.33 ± 0.04 mV; P = 0.07). CONCLUSION: Motor unit discharge rates determined during a brief, submaximal contraction were decreased in the potentiated human tibialis anterior muscle.

Platelet binding and activity of a factor VIIa variant with enhanced tissue factor independent activity.

BACKGROUND AND OBJECTIVES: Platelet binding and activity play important roles in the efficacy of factor VIIa (FVIIa) as a bypassing agent for hemophilia treatment. An analog of FVIIa with increased tissue factor (TF)-independent activity, NN1731, has been produced by introducing three amino acid changes in the protease domain. NN1731 has a conformation similar to TF-bound FVIIa, even in the absence of TF. This results in much greater intrinsic proteolytic activity, but similar activity in the presence of TF. OBJECTIVES: We hypothesized that these changes would not alter binding to platelets or phospholipid, a characteristic thought to be localized to the Gla domain. The goal of the current work was to compare platelet binding and activity of NN1731 and wild-type FVIIa. METHODS/RESULTS: FVIIa and NN1731 bound identically to phospholipid vesicles as assessed by both activity assays and electrophoretic quasielastic light scattering techniques. However, NN1731 bound to a greater number of sites on activated platelets than FVIIa, as assessed by flow cytometry. Removal of the Gla domain abolished binding of both FVIIa and NN1731. Inhibition of the active site did not reduce NN1731 binding to the level of FVIIa. When corrected for the amount of protein bound, NN1731 had greater activity than FVIIa on platelet surfaces. CONCLUSIONS: While the Gla domain is essential for FVIIa binding to platelets, changes in the protease domain in NN1731 enhanced platelet binding as well as proteolytic activity. Features in addition to lipid composition appear to contribute to binding of rFVIIa and, especially, NN1731 to platelets.

The surface EMG-force relationship during isometric dorsiflexion in males and females.

This study compared the tibialis anterior (TA) surface electromyographic (sEMG) to force relationship for males and females. One-hundred participants (50 males and 50 females) performed three isometric contractions at 20, 40, 60, 80, and 100% of maximal voluntary contraction (MVC) in an apparatus designed to isolate the action of the dorsiflexors. The sEMG signal was amplified (1000x), band-pass filtered (10-500 Hz), and sampled at 2048 Hz. The load cell signal was low-pass filtered at 100 Hz and sampled at the same rate. Males were stronger than females (p < 0.05). However, there was no significant difference in the root-mean-square (RMS) amplitude of the sEMG signal between males and females (p < 0.05). Both groups exhibited a quadratic increase in the RMS across force levels (p < 0.05). The mean power frequency (MNF) of the sEMG signal for males was greater than for females (p < 0.05). Males and females exhibited a linear increase in MNF means up to 80% of MVC (p < 0.05). Between 80 and 100% MVC, the frequency values for the females plateaued while males showed a decrease (p < 0.05). The magnitude of the difference in MNF between males and females was consistent with the observation that males have greater type II muscle fiber diameters. In general, the pattern of means for RMS and MNF between males and females revealed no differences between groups in the sEMG-force relationship. We therefore conclude that there are no differences between males and females in the gradation of muscle force.

Modeling investigation of learning a fast elbow flexion in the horizontal plane--prediction of muscle forces and motor units action.

Experimental investigation of practicing a dynamic, goal-directed movement reveals significant changes in kinematics. Modeling can provide insight into the alterations in muscle activity, associated with the kinematic adaptations, and reveal the potential motor unit (MU) firing patterns that underlie those changes. In this paper, a previously developed muscle model and software (Raikova and Aladjov, Journal of Biomechanics, 35, 2002) have been used to investigate changes in MU control, while practicing fast elbow flexion to a target in the horizontal plane. The first trial (before practice) and the last trial (after extensive practice) of two subjects have been simulated. The inputs for the simulation were the calculated external moments at the elbow joint. The external moments were countered by the action of three flexor muscles and two extensor ones. The muscles have been modeled as a mixture of MUs of different types. The software has chosen the MU firing times necessary to accomplish the movement. The muscle forces and MUs firing statistics were then calculated. Three hypotheses were tested and confirmed: (1) peak muscle forces and antagonist co-contraction increase during training; (2) there is an increase in the firing frequency and the synchronization between MUs; and (3) the recruitment of fast-twitch MUs dominates the action.

Recombinant human factor VIIa (rFVIIa) can activate factor FIX on activated platelets.

The studies reported here show that factor (F)VIIa can activate factor (F)IX on activated platelets in the absence of tissue factor. Both FIX and FIXa bind to the activated platelet surface with a K(d) of 8 nM and 2 nM, respectively. With factor (F)VIIIa, FIXa binds more tightly to platelets (K(d) 0.6 nM). At rFVIIa concentrations < 100 nm, no direct binding to the activated platelet surface can be detected with electrophoretic light scattering. However, in the presence of FIX, rFVIIa binding to platelets at concentrations as low as 10 nm rFVIIa can be detected. This is reflected by a decrease in the FIX K(d) from 8 to 1.6 nM. When rFVIIa is added to activated platelets in the presence of both FIX and FVIIIa, the K(d) for FIX decreases to 0.6, suggesting that rFVIIa activates FIX on the surface of activated platelets in the absence of tissue factor. The activation of FIX by FVIIa on activated platelets can also be demonstrated by a functional assay for FIXa. These data show that pharmacological doses of rFVIIa result in the direct activation of FIX by rFVIIa to form additional tenase complexes ultimately resulting in improved thrombin generation. These results may explain, at least in part, the mechanism of action of rFVIIa in hemorrhagic conditions seen in otherwise normal patients who develop an acquired coagulopathy due to trauma, surgery or a variety of other events in which rFVIIa has been found to be effective.

Multidirectional neck strength and electromyographic activity for normal controls.

OBJECTIVE: To assess the multidirectional force and indwelling electromyographic activity during maximal effort isometric actions of the neck. DESIGN: A descriptive study involving maximal effort isometric actions of the neck and bilateral electromyographic activity. BACKGROUND: This study extends previous efforts to assess the isometric strength of the neck, but with greater precision with respect to the intermediate angles between the frontal and sagittal planes. METHODOLOGY: Participants (n = 18) generated a maximal isometric force in twelve directions in the horizontal plane. All exertions were realized in neutral position. A load cell measured forces and intramuscular fine-wire electrodes were used to record the bilateral electromyographic activity of the sternocleidomastoid, scalenus medius, trapezius (middle fibers), semispinalis capitis, and splenius capitis. RESULTS: Strength in the anterolateral directions were similar, but exhibited right lateral dominance in extension. The sternocleidomastoid and trapezius (middle fibers) exhibited bilateral symmetry while the scalenus medius, semispinalis capitis, and splenius capitis did not. Furthermore, the agonist, synergist, and antagonist action of the individual muscles was clear. The direction of force that resulted in the greatest electromyographic activity was consistent with what has been shown in anatomy texts. This was not true for the scalenus medius. We showed that the scalenus medius contributes to extension, with synergistic activity in the lateral bending direction. CONCLUSIONS: The greater precision revealed novel information about the isometric strength of the neck and its musculature.

Reliability of a new measure of H-reflex excitability.

OBJECTIVE: This study examined the intraclass reliability of different measures extracted from Hoffmann reflex (H-reflex) stimulus-response curve that are used to assess neuromuscular excitability. The following measures were compared: (1) the peak-to-peak amplitude of the H-reflex at a stimulus intensity associated with 5% of the maximum M-wave; (2) the slope of the regression line of the H-reflex stimulus-response curve; and (3) the peak of the first derivative of the H-reflex stimulus-response curve, a new measure introduced in this paper. METHODS: The H-reflex was elicited in the soleus for 24 subjects (12 males and 12 females) on 5 separate days. Vibration was applied to the Achilles tendon prior to stimulation to test the sensitivity of the measures on test day 4. The stimulus intensity was gradually increased from below the threshold for an H-reflex response to above the maximum M-wave (Mmax) response. The means of 5 evoked potentials at each intensity level were used to create both the H-reflex and M-wave stimulus-response curves for each subject across test days. Determination of reliability involves the consideration of both the stability and consistency of the measures. A repeated measures analysis of variance evaluated the stability of the group means across test sessions. The consistency of scores within individuals was determined by calculating the intraclass correlation coefficient (ICC). Calculation of the 95% confidence interval of estimation was used to assess significant differences between ICCs. RESULTS: The H-reflex measures were both stable and consistent across the first 3 test days. Achilles tendon vibration resulted in a profound reduction (59-70%) on test day 4, and then there was a return to baseline levels on test day 5. The ICC for H-reflex at a stimulus intensity associated with 5% of the maximum M-wave was 0.85. The ICC for the slope of the regression line of the H-reflex stimulus-response curve was 0.79, while it was 0.89 for the peak of the first derivative of the H-reflex stimulus-response curve. However, there was no statistical significance (P>0.05) between the 3 EMG measures of the H-reflex arc. Maximum M-wave amplitude had an ICC of 0.96 attesting to careful methodological controls. CONCLUSIONS: The peak of the first derivative of the H-reflex stimulus-response curve was shown to have comparable sensitivity and reliability as other more established measures. SIGNIFICANCE: The first derivative of the H-reflex stimulus-response curve provides the rate of change, rather than amplitude, making it a robust measure of reflex arc excitability. The higher ICC for the first derivative offers greater statistical power, which is of practical significance.

Analyzing fibrin clot structure using a microplate reader.

Fibrin clot structure studies are often performed using optical methods. For example, the clot's fiber structure can be assessed by measuring light scattering as a function of wavelength. From these measurements, one can calculate the mass/length ratio (mu), a relative measure of fibrin thickness. Fiber thickness has important functional correlates in terms of clot stability and resistance to fibrinolysis. Typically, measurements to calculate mass/length ratios are carried out on high-end spectrophotometers. However, limitations of this instrument include the large sample volume required and the inability to read multiple samples at one time. To circumvent these limitations, a plate-reading spectrophotometer is more commonly used to monitor clot formation; increases in absorbance indicate clot formation, while decreases indicate clot lysis. However, it is unclear whether plate-reading spectrophotometers can be used to quantitatively evaluate fibrin fiber structure. In the current study, we compared spectrophotometric analysis of fibrin gels on single-sample and plate-reading spectrophotometers. Results show that a plate-reading spectrophotometer does not give accurate measurements of the fiber mass/length ratio. However, the plate-reading spectrophotometer can provide a qualitative measure of fiber structure for both purified fibrinogen and plasma. We suggest that plate-reading spectrophotometers can provide a convenient, rapid, and inexpensive means of analyzing fibrin clot structure.

Neural adaptations to fatigue: implications for muscle strength and training.

PURPOSE: This paper investigates the neural mechanisms responsible for the increase in strength that occurs during serial isometric contractions. METHODS: A three-session design was used. Thirteen subjects (N = 13) were asked to perform five maximal isometric elbow extension strength trials to serve as baseline. After a 5-min rest, the subjects were administered a 30-trial fatigue protocol. This process was repeated two more times at 2-wk intervals. Elbow extension torque and surface electromyography (EMG) of the triceps and biceps brachii were monitored concurrently. The criterion measures were elbow extension torque, root-mean-square EMG amplitude, and mean power frequency (MPF). RESULTS: Intraclass reliability ranged from good to excellent. Within each experimental session, the fatigue protocol resulted in a decrease in maximal isometric elbow extension torque as well as biceps and triceps EMG amplitude and MPF (P < 0.05). However, the mean of the 30 trials and the magnitude of the linear decrease in elbow extension torque increased across the three sessions (P < 0.05). Biceps and triceps EMG amplitude increased and MPF decreased as the number of sessions increased (P < 0.05). CONCLUSIONS: These findings suggest that the fatigue protocol served as a training stimulus to down regulate motor-unit firing frequency.

Location of the platelet binding site in zymogen coagulation factor IX.

The assembly of the tenase complex on the surface of the platelet is an essential step in maintaining normal hemostasis as evidenced by the serious hemorrhagic diathesis associated with either factor IX (FIX) or factor VIII deficiencies. Understanding the regions and or residues of FIX crucial for proper binding to platelets has important clinical implications. The ability of FIX to bind activated platelets in the presence of 4 mmol/l CaCl2 was examined using electrophoretic light-scattering experiments. Wild-type FIX binds to activated platelets with dissociation constant Kd = 7.9 nmol/l. Activated FIX binds to activated platelets with Kd = 2 nmol/l. Activated factor VII does not bind activated platelets at physiological concentrations. The Gla domain of FIX is important for the binding of FIX to activated platelets since a chimera with a factor VII (FVII) template and FIX Gla [FVII(FIXGla)] has Kd = 9.6 nmol/l, and a chimera with a FVII template and FIX Gla, A and the first epidermal growth factor domain (EGF1) [FVII(FIXGla,A,EGF1)] has Kd = 9.7 nmol/l, but a chimera with a FIX template and a FVII Gla [FIX(FVIIGla)] does not bind activated platelets. Altering the fifth residue of FIX from a lysine to an alanine (Lys5<--Ala) abolishes the mutant from binding to collagen but does not affect FIX binding to the activated platelet (Kd = 9.8 nmol/l). Point mutations involved with residues 4 and 5 (Gly4<--Phe and Lys5<--no residue), residue 9 (Phe9<--Ala), residue 10 (Val10<--Lys) and residues 9-11 (Phe9<--Met, Val10<--Lys, Glu11<--Lys) do not bind to activated platelets.

Training-related changes in the maximal rate of torque development and EMG activity.

This study monitored the effects of a short-term elbow flexor training program on surface electromyographic (SEMG) spike activity. The experimental paradigm consisted of three test sessions separated by 2-week intervals. At the beginning of each session, participants (N=13) performed five maximal effort isometric contractions of the elbow flexors to serve as baseline. After 5 min of rest, the participants then engaged in a 30-trial isometric fatigue protocol during which maximal elbow flexion torque was measured with a load-cell, and the maximal rate of change in the torque (dtau/dt(max)) was obtained from the differentiated torque-time curve. Bipolar electrodes were used to monitor the SEMG spike activity of the biceps brachii. Mean spike amplitude (MSA) and mean spike frequency (MSF) were calculated for the torque development and constant-torque phases of the isometric contraction, termed Segment 1 and Segment 2, respectively. Mean power frequency (MPF) was also calculated for Segment 2. The five baseline contractions of the second and third sessions were compared with those of the first session and analyzed for training-related changes. Training increased dtau/dt(max) but failed to change maximal elbow flexion torque or MSA. However, there was an increase in the MSF during the torque development phase of the contraction (Segment 1). Both MSA and MSF were greatest during the constant-torque phase of the isometric contraction (Segment 2). There was a strong linear correlation (r=0.90, P<0.05) between MSF and MPF during (Segment 2). We hypothesize that the increase in dtau/dt(max) is due to enhanced motor-unit rate-coding. The demonstrated correlation between MSF and MPF measures will allow investigators to use spike analysis to examine the frequency content of the SEMG signal under non-stationary conditions.

The reversal of antagonists facilitates the peak rate of tension development.

OBJECTIVE: The present study was designed to test the effects of the reversal of antagonists on the peak rate of tension development (dF/dt(max)) of the elbow extensors. DESIGN: Experimental, with matched controls. SETTING: A biomechanics research laboratory. PARTICIPANTS: Twenty-six healthy women without a history of upper extremity injury or neurologic disorder, randomly assigned to experimental (n = 13) or control (n = 13) groups. INTERVENTIONS: Two groups of healthy subjects followed identical exercise protocols, except that the control group performed maximal isometric contractions of the elbow extensors and the experimental group executed a maximal isometric elbow flexion contraction immediately before a maximal elbow extension contraction. Both groups performed 5 cycles of a 2-second contraction with 22-second rest periods between agonist muscle contractions and were evaluated at 4 test sessions spaced 2 weeks apart. MAIN OUTCOME MEASURES: All measurements were done with the shoulder and elbow at 90 degrees of flexion in the sagittal plane to ensure reproducibility. A load cell was used to measure elbow extension moment and to calculate the peak rate of tension development (dF/dt(max)). Biceps and triceps brachii surface electromyographic activity was monitored concurrently. The electromyographic measures were mean spike (peak-to-peak) amplitude and mean spike frequency of the biceps and triceps brachii activity. RESULTS: Intraclass dF/dt(max) and electromyographic reliability was good (r > or = .72) in both groups. Because biceps electromyographic measures were considerably less reliable (r < or = .53), they were not included in our analysis. While dF/dt(max) increased quadratically in both groups (p <.05), the experimental group was on average 36.1Nm. s(-1) (63%) greater across sessions 2 to 4 (p <.05). In contrast, triceps electromyographic activity did not differ significantly between groups (p >.05). The means averaged across groups exhibited a quadratic increase from session 1 to session 4: 91microV or 48% for mean spike amplitude (p <.05) and 7Hz or 16% for mean spike frequency (p <.05). CONCLUSIONS: The greater dF/dt(max) for the experimental group was not associated with increased electromyographic activity. The experimental group appeared to use the biomechanic properties of the pretensioned extensor muscle-tendon complex, rather than neurologic biasing, to accomplish its power gains.

Practicing a maximal performance task: a cooperative strategy for muscle activity.

The effect of practice on predicting elbow flexion movement time was studied. Participants (N = 18) performed 400 elbow flexion trials to a target in the horizontal plane. The trials were distributed equally over four sessions. The goal was to decrease the movement time (MT) for the same degree of accuracy. The electromyographic (EMG) activity of the biceps and triceps brachii was monitored with standard Beckman Ag/AgCl surface electrodes. The EMG measures formed two variable sets within one prediction equation. One variable set was composed of the onset of muscle activity relative to the start of movement (motor time) and the duration of muscle activity. The other variable set consisted of the mean amplitude value of the entire burst and of the first 30 ms (Q30) of activity. As the maximal speed of limb movement increased, the duration of muscle activity (motor time and EMG duration) decreased, and the magnitude of muscle activity (MAV and Q30) increased. Most of the change in the duration of muscle activity occurred in Session 1, while the magnitude of muscle activity continued to increase until Session 3. Multiple regression analysis revealed a cooperative strategy between the magnitude and duration of muscle activity. Early in learning, participants adjusted the magnitude of muscle activity to increase limb movement speed. As practice continued, alterations in the duration of muscle activity became more important, while the magnitude changes were less involved. Late in learning, both dimensions of muscle activity were used to decrease MT. We suggest that the interplay between the magnitude and duration of muscle activity may be due to: (a) cognitive factors related to the division of attention in a motor skill, (b) an increase in the frequency of motor unit firing that affects both dimensions of muscle activity, or (c) some combination of (a) and (b).

Comparison of granulocyte colony-stimulating factor (G-CSF)--mobilized peripheral blood progenitor cells and G-CSF--stimulated bone marrow as a source of stem cells in HLA-matched sibling transplantation.

HLA-identical bone marrow or stem cell transplantation from a sibling is the preferred treatment for patients with chronic myelogenous leukemia, bone marrow failure syndromes, relapsed acute leukemia, and specific inborn errors of metabolism. Several groups have shown that granulocyte colony-stimulating factor (G-CSF)--mobilized peripheral blood progenitor cells (PBPCs) obtained from HLA-matched siblings are effective in reconstitution of marrow function after marrow ablative conditioning therapy. To evaluate whether G-CSF treatment before bone marrow harvest leads to enhanced recovery of PBPC counts and recovery from limited graft-versus-host disease (GVHD), we assessed the outcome of a sequential cohort of patients treated identically and then given either G-CSF--mobilized PBPCs or G-CSF--stimulated bone marrow from HLA-identical siblings. We show that the time to neutrophil engraftment is identical in the 2 cohorts, whereas platelet engraftment is earlier with the use of PBPCs. The incidence of acute GVHD was decreased, and that of chronic GVHD significantly decreased, in the group receiving bone marrow. Overall survival was not different between the 2 groups. Thus, G-CSF--stimulated bone marrow offers a source of stem cells that allows for early neutrophil engraftment with a decreased risk of GVHD.

CD34+ selection of hematopoietic blood cell collections and autotransplantation in lymphoma: overnight storage of cells at 4 degrees C does not affect outcome.

The purpose of this study was to investigate whether storing mobilized peripheral blood progenitor cell (PBPC) collections overnight before CD34+ selection may delay platelet count recovery after high-dose chemotherapy and CD34+-enriched PBPC re-infusion. Lymphoma patients underwent PBPC mobilization with cyclophosphamide 4 g/m2 i.v. and G-CSF 10 microg/kg/day subcutaneously. Patients were prospectively randomized to have each PBPC collection enriched for CD34+ cells with the CellPro CEPRATE SC System either immediately or after overnight storage at 4 degrees C. Thirty-four patients were randomized to overnight storage and 34 to immediate processing of PBPC; 15 were excluded from analysis due to tumor progression or inadequate CD34+ cell mobilization. PBPC from 23 patients were stored overnight, while 30 subjects underwent immediate CD34+ selection and cryopreservation. Median yield of CD34+ enrichment was 43.6% in the immediate processing group compared to 39.1% in the overnight storage group (P = 0.339). Neutrophil recovery >500 x 10(9)/l occurred a median of 11 days (range 9-16 days) in the overnight storage group compared to 10.5 days (range 9-21 days) in the immediate processing group (P = 0.421). Median day to platelet transfusion independence was 13 (range 7-43) days in the overnight storage group vs 13.5 (range 8-35) days in those assigned to immediate processing (P = 0.933). We conclude that storage of PBPC overnight at 4 degrees C allows pooling of consecutive-day collections resulting in decreased costs and processing time without compromising neutrophil and platelet engraftment after infusion of CD34+-selected progenitor cells. Bone Marrow Transplantation(2000) 25, 559-566.