Use of Ex Vivo Normothermic Perfusion for Quality Assessment of Discarded Human Donor Pancreases.

A significant number of pancreases procured for transplantation are deemed unsuitable due to concerns about graft quality and the associated risk of complications. However, this decision is subjective and some declined grafts may be suitable for transplantation. Ex vivo normothermic perfusion (EVNP) prior to transplantation may allow a more objective assessment of graft quality and reduce discard rates. We report ex vivo normothermic perfusion of human pancreases procured but declined for transplantation, with ABO-compatible warm oxygenated packed red blood cells for 1-2 h. Five declined human pancreases were assessed using this technique after a median cold ischemia time of 13 h 19 min. One pancreas, with cold ischemia over 30 h, did not appear viable and was excluded. In the remaining pancreases, blood flow and pH were maintained throughout perfusion. Insulin secretion was observed in all four pancreases, but was lowest in an older donation after cardiac death pancreas. Amylase levels were highest in a gland with significant fat infiltration. This is the first study to assess the perfusion, injury, as measured by amylase, and exocrine function of human pancreases using EVNP and demonstrates the feasibility of the approach, although further refinements are required.

Comparative clinical- and cost-effectiveness of candesartan and losartan in the management of hypertension and heart failure: a systematic review, meta- and cost-utility analysis.

The UK National Health Service (NHS) currently spends in excess of £250 million per annum on angiotensin II receptor blockers (ARBs) for the treatment of hypertension and heart failure; with candesartan currently dominating the market. With the recent introduction of generic losartan, we set out to directly compare the branded market leader to its now cheaper alternative. The primary objectives were to compare the blood pressure (BP) lowering efficacy and cardiovascular outcomes of candesartan and losartan in the treatment of essential hypertension and chronic heart failure, respectively. The secondary objective was to model their comparative incremental cost-effectiveness in a UK NHS setting. The Cochrane Central Register of Controlled Trials (Cochrane Library 2009, issue 2), which contains the Hypertension and Heart Group's specialist register, Medline (1950-February 2010), and Embase (1980-February 2010) were included in the search strategy. Selection criteria were randomised studies of candesartan versus losartan in adults (> 18 years). The main outcome measures were as follows: Hypertension: mean change from baseline in trough (24 h postdose) systolic and diastolic BP. Heart failure: composite of cardiovascular death and hospital admission for management of heart failure. Two reviewers applied inclusion criteria, assessed trial quality, and extracted data. Eight (three of which met inclusion criteria) and zero trials compared candesartan directly with losartan in the treatment of hypertension and heart failure, respectively. A between-treatment difference of -1.96 mmHg [95% confidence interval (CI) -2.40 to -1.51] for trough diastolic BP and -3.00 mmHg (95% CI -3.79 to -2.22) for trough systolic BP in favour of candesartan was observed. Based on this differential, a 10-year Markov model estimates the cost per quality-adjusted life-year gained to exceed £40,000 for using candesartan in place of generic losartan. Candesartan reduces BP to a slightly greater extent when compared with losartan, however, such difference is unlikely to be cost-effective based on current acquisition costs, perceived NHS affordability thresholds and use of combination regimens. We could find no robust evidence supporting the superiority of candesartan over losartan in the treatment of heart failure. We therefore recommend using generic losartan as the ARB of choice which could save the UK NHS approximately £200 million per annum in drug costs.

High-dose oral vitamin D3 supplementation in the elderly.

SUMMARY: Daily dosing with vitamin D often fails to achieve optimal outcomes, and it is uncertain what the target level of 25-hydroxyvitamin D should be. This study found that large loading doses of vitamin D(3) rapidly and safely normalize 25OHD levels, and that monthly dosing is similarly effective after 3-5 months. With baseline 25OHD > 50 nmol/L, vitamin D supplementation does not reduce PTH levels. INTRODUCTION: There is concern that vitamin D supplementation doses are frequently inadequate, and that compliance with daily medication is likely to be suboptimal. METHODS: This randomized double-blind trial compares responses to three high-dose vitamin D(3) regimens and estimates optimal 25-hydroxyvitamin D (25OHD) levels, from changes in parathyroid hormone (PTH), and procollagen type I amino-terminal propeptide (P1NP) in relation to baseline 25OHD. Sixty-three elderly participants were randomized to three regimens of vitamin D supplementation: a 500,000-IU loading dose; the loading dose plus 50,000 IU/month; or 50,000 IU/month. RESULTS: The Loading and Loading + Monthly groups showed increases in 25OHD of 58 +/- 28 nmol/L from baseline to 1 month. Thereafter, levels gradually declined to plateaus of 69 +/- 5 nmol/L and 91 +/- 4 nmol/l, respectively. In the Monthly group, 25OHD reached a plateau of ~80 +/- 20 nmol/L at 3-5 months. There were no changes in serum calcium concentrations. PTH and P1NP were only suppressed by vitamin D treatment in those with baseline 25OHD levels <50 and <30 nmol/L, respectively. CONCLUSIONS: Large loading doses of vitamin D(3) rapidly and safely normalize 25OHD levels in the frail elderly. Monthly dosing is similarly effective and safe, but takes 3-5 months for plateau 25OHD levels to be reached.

Prothrombotic and inflammatory effects of intravenous administration of human immunoglobulin G in dogs.

BACKGROUND: Intravenous administration of human immunoglobulin G (hIVIgG) has been suggested to potentiate thromboembolism in dogs, but supportive scientific reports are lacking. OBJECTIVES: To determine if hIVIgG therapy promotes hypercoagulability and inflammation in dogs. ANIMALS: Twelve healthy Beagle dogs. METHODS: Prospective, experimental trial. An hIVIgG/saline solution was infused IV at 1 g/kg BW over 8 hours to 6 dogs, and physiological saline was infused to the other 6 dogs. Blood samples were drawn before, during, and after infusion for serial measurement of indicators of coagulation and inflammation. Data were analyzed by 2-way repeated measures analysis of variance. RESULTS: Dogs administered hIVIgG developed mildly decreased blood platelet concentrations without thrombocytopenia (median, 200 x 10(3)/microL; range, 150-302 x 10(3)/microL; P < .01), leukopenia (median, 3.5 x 10(3)/microL; range, 20-62 x 10(3)/microL; P < .001), and mildly increased plasma total protein concentrations (median, 6.3 g/dL; range, 5.6-6.7 g/dL; P < .001). Administration of hIVIgG was also associated with increases in fibrin/fibrinogen degradation products in all dogs (either 5 microg/mL or 10 microg/dL), thrombin-antithrombin III complexes (median, 7.2 ng/mL; range, 4.9-14.2 ng/mL; P < .001), and C-reactive protein concentrations (median, 2.5 mg/dL; range, 0.5-4.3 mg/dL; P < .01). CONCLUSION AND CLINICAL IMPORTANCE: Administration of hIVIgG to dogs promotes hypercoagulability and an inflammatory state. This should be further evaluated and considered when using hIVIgG in dogs with IMHA or other prothrombotic conditions.

Leaving hip rotation out of a conventional 3D gait model improves discrimination of pathological gait in cerebral palsy: A novel neural network analysis.

BACKGROUND: Complex clinical gait analysis results can be expressed as single number gait deviations by applying multivariate processing methods. The original Movement Deviation Profile (MDP) quantifies the deviation of abnormal gait using the most trusted nine dynamic joint angles of lower limbs. RESEARCH QUESTION: Which subset of joint angles maximises the ability of the MDP to separate abnormal gait from normality? What is the effect of using the best subset in a large group of patients, and in individuals? METHODS: A self-organising neural network was trained using normal gait data from 166 controls, and then the MDP of 1923 patients with cerebral palsy (3846 legs) was calculated. The same procedure was repeated with 511 combinations of the nine joint angles. The standardised distances of abnormal gait from normality were then calculated as log-transformed Z-scores to select the best combination. A mixed design ANOVA was used to assess how removing the least discriminating angle improved the separation of patients from controls. The effect of using the optimal subset of angles was also quantified for each individual leg by comparing the change in MDP to the independent FAQ levels of patients. RESULTS: Removal of hip rotation significantly (p<0.0005) increased the separation of the patient group from normality (ΔZ-score 0.24) and also at FAQ levels 7-10 (ΔZ-score 0.38, 0.27, 0.22, 0.14). The MDP of individual patients changed in a wider range of -4.65 to 1.12 Z-scores and their change matched their independent FAQ scores, with less functional patients moving further from, and more functional patients moving closer to normality. SIGNIFICANCE: In existing gait databases we recommend excluding hip rotation from data used to calculate the MDP. Alternatively, the calculation of hip rotation can be improved by post-hoc correction, but the ultimate solution is to use more accurate and reliable models of hip rotation.

3D printed structures for modeling the Young's modulus of bamboo parenchyma.

Bamboo is a sustainable, lightweight material that is widely used in structural applications. To fully develop micromechanical models for plants, such as bamboo, the mechanical properties of each individual type of tissue are needed. However, separating individual tissues and testing them mechanically is challenging. Here, we report an alternative approach in which micro X-ray computed tomography (µ-CT) is used to image moso bamboo (Phyllostachys pubescens). The acquired images, which correspond to the 3D structure of the parenchyma, are then transformed into physical, albeit larger scale, structures by 3D printing, and their mechanical properties are characterized. The normalized longitudinal Young's moduli of the fabricated structures depend on relative density raised to a power between 2 and 3, suggesting that elastic deformation of the parenchyma cellular structure involves considerable cell wall bending. The mechanical behavior of other biological tissues may also be elucidated using this approach. STATEMENT OF SIGNIFICANCE: Bamboo is a lightweight, sustainable engineering material widely used in structural applications. By combining micro X-ray computed tomography and 3D printing, we have produced bamboo parenchyma mimics and characterized their stiffness. Using this approach, we gained insight into bamboo parenchyma tissue mechanics, specifically the cellular geometry's role in longitudinal elasticity.

Application of growth factor stimulants improves cytogenetic analysis of chronic myeloproliferative disorder patients without alteration to cell lineage or clonality.

Conventional cytogenetic methods rely on culturing bone marrow aspirates to obtain suitable and sufficient mitotic figures for G-banded analysis. Samples from patients with chronic myeloproliferative disorders (CMPD) often have increased failure rates due to reduced growth and poor morphology, all of which hamper the conventional karyotyping investigation. The application of growth factor (GF) stimulants to bone marrow aspirates has been shown to yield significant increases in both the quality and quantity of bone marrow metaphases obtained in 53 CPMD patient samples. All cultures were stimulated using the conditioned supernatant from the human bladder carcinoma cell line 5637, which contains IL-3, IL-6, and G-CSF. Results were assessed qualitatively on G-banded preparations and quantitatively by mitotic index (MI = % dividing cells). To assess whether the application of GF stimulants leads to clonal selection, culture samples from 15 patients were analyzed by fluorescence in situ hybridization, which supported the theory that clonal selection remains unaltered in GF-stimulated cultures. In addition to this immunophenotyping of cells, we demonstrated the lineage of cells propagated under these conditions. Cell markers were chosen to characterize B-lymphoid, T-lymphoid, myeloid, and primitive cell types. Results indicated that T cells were maintained in culture and B-lymphoid markers remained negative. In the myeloid subset, there was an overall reduction in the pan-myeloid markers. We believe this represents the loss of terminally differentiated cells (e.g., neutrophils) in culture. Overall, the study clearly demonstrates that the application of GF stimulants does not alter clonality or cell lineage propagated in these samples and is therefore suitable for application in diagnostic cytogenetic laboratories.

Structural and chemical changes in ultra-high-molecular-weight polyethylene due to gamma radiation-induced crosslinking and annealing in air.

Ultra-High-Molecular-Weight-Polyethylene (UHMWPE) is the material of choice for one of the articulating surfaces in many total joint replacements, notably hip and knee prostheses. The various methods used by the orthopaedic biomaterials industry to sterilize and anneal UHMWPE components, and the resulting oxidation and crosslinking, affect the mechanical wear resistance properties in ways still unknown at the microscopic and molecular levels. Transmission electron microscopy and chemical pyrolysis were used to quantify crosslinking induced by gamma irradiation and annealing in air. Changes in lamellar stacking and the amount of crosslinking suggest two types of crosslinking: relatively unstable crosslinks in the amorphous region initially resulting from gamma irradiation which are later replaced by more thermally stable crosslinks resulting from rearrangements at the annealing temperature. Lamellar mobility, the ability of crystalline lamellae to flow in the material, is enhanced during the transition from one type of bond to the other, and this appears to optimize near eight hours of annealing time. Results from decomposition and percent crystallinity measurements provide further support for this theory.

Acute anhydrous ammonia intoxication in cattle.

Intoxication with anhydrous ammonia (AA) is a common occupation-related health problem affecting farmers and fertilizer applicators, however, very few descriptions of animal exposure to this toxicant exist. Thieves entered a feedlot and damaged a liquid AA storage tank, resulting in the accidental leakage of gas from a valve. Overnight, 12 cattle were found dead, with a total mortality of 64 cattle out of 260 on the premises dying or euthanized in < 1 week. Signs of affected cattle included blindness, drooling, inappetence, respiratory distress, recumbency, and death. Two cattle were submitted live for diagnostic evaluation 3 days after initial exposure. Gross lesions included corneal ulcers, fibrinonecrotizing rhinitis, hemorrhages within the nasal sinuses, and anterioventral bronchopneumonia. Microscopic lesions consisted predominantly of degeneration and necrosis of superficial epithelium lining nasal passages, trachea, and pulmonary airways. This case illustrates the acute AA effects on cattle, which is likely to be an increasingly encountered problem because of on-farm storage of AA and its access by illicit drug manufacturers.

Microsatellite analysis of posttransplant lymphoproliferative disorders: determination of donor/recipient origin and identification of putative lymphomagenic mechanism.

The genetic mechanisms that give rise to posttransplant lymphoproliferative disorders (PTLDs) are not well established, yet previous studies have focused or) the role of immunosuppression and EBV infection. We investigated whether microsatellite analysis could: (a) determine the recipient/donor origin of the tumor; and (b) document novel genetic alterations in PTLDs, i.e., microsatellite instability. We characterized seven cases of PTLD (five B-cell and two T-cell non-Hodgkin's lymphomas) in which donor allograft tissue, normal recipient tissue, and tissue from the PTLDs were available. In each case, six microsatellite loci were analyzed. Five cases were of host origin (three B-cell and two T-cell lymphomas). The two cases of donor origin were B-cell lymphomas. Multiple loci showed microsatellite instability in two cases of host-derived T-cell non-Hodgkin's lymphomas (28% of PTLDs). These findings show that microsatellite analysis may be used to determine the host or donor origin of PTLDs and suggest for the first time that defective DNA mismatch repair may be an underlying genetic mechanism of lymphomagenesis in some cases of PTLD.

Megakaryocytes modulate osteoblast synthesis of type-l collagen, osteoprotegerin, and RANKL.

We have previously reported evidence that megakaryocytes may play a role in bone remodeling, possibly by interactions with cells at the bone surface. To investigate the direct effects of megakaryocytes on osteoblasts, maturing megakaryocytes (CD61 positive cells) were isolated and added to cultures of human osteoblasts. Osteoblasts alone and osteoblasts treated with CD61-negative (non-megakaryocytic) cells were used as control cultures. After 48 h in culture, megakaryocytes were removed and osteoblasts immunolocalized for type-1 collagen, osteoprotegerin (OPG), and RANKL expression. Similar cultures were used for RNA extraction with mRNA for Col 1A1, OPG, and RANKL in osteoblasts measured quantitatively by RT-PCR. Osteoblasts cultured alone showed high levels of expression of collagen with 74% (+/-7) of cells staining positively. When cultured with megakaryocytes, the number of positively staining cells remained similar but the intensity of expression was increased 1.54-fold (P < 0.02). OPG was expressed by 32% (+/-6.3) of osteoblasts increasing to 51% (+/-5.5) when cultured in the presence of megakaryocytes (P < 0.01) with a 1.63-fold increase in intensity of expression (P < 0.01). In contrast, osteoblasts cultured with megakaryocytes showed suppression of RANKL expression; 35.6% (+/-5.8) of osteoblasts cultured alone stained positively decreasing to 24.3% (+/-5.3) with a 1.6-fold diminished intensity of expression (P < 0.02). Osteoblasts co-cultured with CD61-negative cells showed no differences in collagen, OPG, or RANKL expression levels compared to osteoblasts cultured alone. mRNA data supported these findings with a 3.1-fold increase in Col 1A1 expression in megakaryocyte-treated cultures compared to controls (P < 0.02). Low-level OPG mRNA expression increased 8.14-fold in osteoblasts cultured in the presence of megakaryocytes (P < 0.01), while RANKL expression was suppressed 3.3-fold (P < 0.02). These results demonstrate that in vitro, megakaryocytes have direct effects on osteoblastic production of factors affecting both bone formation and resorption. These data provide further evidence that megakaryocytes may play an important role in bone remodeling.

Visual search and coordination changes in response to video and point-light demonstrations without KR.

The authors examined the observational learning of 24 participants whom they constrained to use the model by removing intrinsic visual knowledge of results (KR). Matched participants assigned to video (VID), point-light (PL), and no-model (CON) groups performed a soccer-chipping task in which vision was occluded at ball contact. Pre- and posttests were interspersed with alternating periods of demonstration and acquisition. The authors assessed delayed retention 2-3 days later. In support of the visual perception perspective, the participants who observed the models showed immediate and enduring changes to more closely imitate the model's relative motion. While observing the demonstration, the PL group participants were more selective in their visual search than were the VID group participants but did not perform more accurately or learn more.

The histologic spectrum of hepatic cat scratch disease. A series of six cases with confirmed Bartonella henselae infection.

Cat scratch disease (CSD), a common cause of regional lymphadenitis, has been linked to Bartonella henselae infection. Although rare, dissemination with hepatic involvement has been documented. Six cases of hepatic CSD were retrieved and probed for B. henselae DNA. Hematoxylin-eosin, trichrome, methenamine silver, Gram, Ziehl-Neelsen, and Warthin-Starry stained slides were reviewed by all authors, and the histologic findings were recorded. B. henselae infection was confirmed in all cases using combined polymerase chain reaction and Southern blot methods. All biopsies contained multiple granulomas, many with characteristic stellate abscesses surrounded by three distinct zones: an inner layer of palisading histiocytes, an intermediate lymphocytic rim, and an outermost zone of fibrosis. One case had noncaseating granulomas with prominent giant cells. Warthin-Starry stains were positive in two. The surrounding parenchyma in all cases contained lymphocytic infiltrates within expanded fibrotic portal triads and adjacent dilated sinusoids. There is a distinct spectrum of histologic changes in hepatic CSD. Although the unit lesion is granulomatous, the organization varies markedly. All cases show predictable nonspecific surrounding parenchymal changes. When multiple hepatic lesions are found in patients with constitutional symptoms suggesting an infectious illness, CSD should be included in the differential diagnosis.

The role of Yersinia enterocolitica and Yersinia pseudotuberculosis in granulomatous appendicitis: a histologic and molecular study.

Granulomatous appendicitis is an enigmatic entity. Purported causes include Crohn's disease, foreign body reactions, sarcoidosis, and infectious agents; however, most cases remain idiopathic. Yersinia enterocolitica (YE) and Y. pseudotuberculosis (YP) have been implicated as causes of appendicitis, ileocolitis, and mesenteric adenitis. The authors examined the potential role of YE and YP in granulomatous appendicitis using histologic and molecular methods. Forty cases of granulomatous appendicitis were evaluated for histologic features including transmural inflammation, number and character of granulomas, and mucosal changes. Twort Gram, Grocott methenamine-silver (GMS), and Ziehl-Neelsen stains were evaluated, and polymerase chain reaction (PCR) analysis was performed to identify pathogenic YP and YE. Twenty-five percent (10 of 40) of the cases were positive for pathogenic Yersinia by PCR (four YE, four YP, and two with both species). Prominent histologic features included epithelioid granulomas with lymphoid cuffing, transmural inflammation with lymphoid aggregates, mucosal ulceration, and cryptitis. One Yersinia-positive case contained mural Gram-negative bacilli; fungal and acid-fast bacilli stains were all negative. Except for one culture-negative case, serologies and cultures were not done or results were unavailable. Two Yersinia-positive patients were diagnosed subsequently with Crohn's disease, suggesting a possible relationship between the two entities. No other patients developed significant sequelae. YE and YP are important causes of granulomatous appendicitis, and Yersinia infection may mimic Crohn's disease. No histologic features distinguish reliably between Yersinia species, or between Yersinia-positive and Yersinia-negative cases. Because special stains and cultures are often not diagnostic, PCR analysis is an excellent technique for the diagnosis of Yersinia.

Prolongation of primate cardiac allograft survival by treatment with ANTI-CD40 ligand (CD154) antibody.

BACKGROUND: We evaluated whether a humanized anti-CD154 antibody (hu5c8) prolongs primate cardiac allograft survival. METHODS: Heterotopic cardiac allografts were performed between MHC class II-mismatched cynomolgus monkeys. Survival was compared between groups treated with a perioperative dosing of hu5c8 (group 1; n=6), sustained dosing with hu5c8 (group 2; n=3), and control regimens (n=4). All recipients received fresh donor-specific transfusions during surgery. RESULTS: Median graft survival was 49 days (range 14 to 56) in group 1 and 106 days (range 56 to 245) in group 2, compared with 5 days (range 5 to 6) for controls (P<0.05 for all comparisons). Lymphocytic infiltrates were often present in hu5c8-treated grafts with stable contractility. Donor-specific mixed lymphocyte reaction was generally preserved. Vasculitis and cellular intimal proliferation were prevalent in rejected grafts but occurred later and were less prevalent in group 2. CONCLUSIONS: Anti-CD154 antibody markedly prolongs the survival of cardiac allografts in primates and is well tolerated. Sustained dosing with hu5c8 yielded improved survival and may be associated with a lower incidence of vascular pathology. We conclude that hu5c8 therapy is an effective approach for inhibiting acute cardiac allograft rejection in primates.

Ductal carcinoma in situ of the breast: reproducibility of histological subtype analysis.

Historically, two major strata of ductal carcinoma in situ (DCIS) have been linked to outcome, the presence or absence of comedo type and size. Our initial approach in classification was dichotomous, often favoring the comedo type with most worrisome implications to foster agreement in diagnosis. We have now tested guidelines that foster agreement in the modified Lagios three-tiered system. Sixteen cases of DCIS were selected, reflecting a spectrum of histological subtypes, with specific inclusion of cases in which consensus in classification using a dichotomous (comedo/noncomedo) scheme would be difficult. Six independent observers reviewed a minimum of five color 35-mm slides from each case at two separate occasions. The aim was to subclassify each case based on architectural pattern, nuclear grade, and presence or absence of tumor necrosis (Modified Lagios Classification, Lagios et al, Cancer 1989). After initial review, emphasizing placement of each case into a high- or low-grade category, there was disagreement in seven cases (44%), confirming our aim to choose cases with uncertain cues for classification. Agreement was achieved in 94% of cases by allowing re-review with emphasis on inclusion of an intermediate-grade category. Our study also suggests that pure micropapillary DCIS and apocrine DCIS warrant independent classification as "special type" DCIS. Our small pilot study suggests that, with adherence to specific criteria, most DCIS cases can be easily and consistently classified into the following five categories: (1) high grade, (2) intermediate grade, (3) low grade, (4) pure or predominantly micropapillary, and (5) pure apocrine. Our six observers independently reached a final concordance of 94% despite selection of cases in which consensus in a dichotomous classification was difficult. This was achieved predominantly by accepting an intermediate category of DCIS with intermediate nuclear features and limited necrosis. Confirmation of the applicability of the Modified Lagios Classification awaits completion of a much larger multi-institutional study in which statistical significance and interobserver variation can be better defined.

A trend towards an increased incidence of chronic graft-versus-host disease following allogeneic peripheral blood progenitor cell transplantation: a case controlled study.

Allogeneic peripheral blood progenitor cell transplantation (alloPBPCT) is increasingly used as an alternative to bone marrow transplantation (alloBMT). Early data suggest that the incidence and severity of acute graft-versus-host disease (GVHD) following alloPBPCT is no higher than that seen with alloBMT, despite the increased number of cytotoxic T cells infused with mobilised blood. We compared 12 patients undergoing alloPBPCT with 12 well-matched alloBMT controls. All patients received identical GVHD prophylaxis. No T cell depletion or CD34 purification was performed. Median engraftment times for neutrophils >0.5 x 10(9)/l and platelets >20 x 10(9)/l were 14 and 12 (alloPBPCT) and 21 and 23 days (alloBMT), respectively (P = 0.0035 and P = 0.002). There was no difference in antibiotic requirements (P = 0.83), platelet support (P = 0.59) or days in hospital (P = 0.51). After alloPBPCT, five patients developed > or =grade II acute GVHD vs five patients after alloBMT (P = 0.99). There was one death (alloBMT) at 100 days and three at 1 year (all due to relapse). There was one death at 100 days with alloPBPCT, and 11 patients remain alive (range 9-21 months) to date. Chronic GVHD occurred in five patients in the PBPC arm and one patient in the BM arm (P = 0.14). This case-controlled analysis indicates that alloPBPCT results in more rapid engraftment kinetics but in no significant difference in transplant-related morbidity or mortality. There is no difference in the incidence of acute GVHD. However, there is a trend towards increased incidence of chronic GVHD in patients allografted with PBPC. Prospective randomised trials are required to determine further the role of alloPBPCT.

Improved yield of peripheral blood progenitor cells on the Cobe Spectra.

The efficiency of leucapheresis using the Cobe Spectra depends on the observation of the operator to monitor the haematocrit (Hct) in the collection line at the recommended Hct of 1%. Cells were harvested by processing 3 x total blood volume (TBV) over 3-4 h. During the harvest the manual Hct and the colony-forming units granulocyte-macrophage (CFU-GM) concentrations were analysed in the collection line in the observed Hct range of 1-7.5%. There was a correlation between the manual and observed Hct (r = 0.97). The CFU-GM concentration was normalised to allow comparison between patients. The increase was statistically significant between 1-2% (P = 0.04) and 1-3% Hct (P = 0.05). The increase in CFU-GM concentration remained at the termination of harvest, indicating that not all available CFU-GM were harvested. The optimum concentration of progenitor cells was found to be at 3% Hct. We postulate that this may permit the collection of sufficient cells on one occasion to allow PBPC autografting in the majority of patients who respond to mobilisation.

Liquid storage of peripheral blood progenitor cells for transplantation.

Peripheral blood progenitor cells (PBPC) were mobilised by recombinant human G-CSF (rhG-CSF) and cyclophosphamide, harvested by apheresis from 9 patients with NHL and stored at 4 degrees C without further manipulation. They were then reinfused after high-dose chemotherapy. We monitored the change in colony-forming units-granulocyte, macrophage (CFU-GM) proliferation as well as plasma glucose, lactate and pH levels over the period of the study. In an attempt to maintain CFU-GM numbers, rhG-CSF was added to storage bags at collection and 48 h later. Our observations suggest that cell viability is well maintained and that CFU-GM numbers rise over the first 48 h of storage before falling rapidly. Metabolic changes cause a fall in the pH and glucose levels with a reciprocal rise in plasma lactate. The addition of rhG-CSF at a concentration of 10 ng/ml to cells in storage showed no detectable benefit. Following storage for 96 h, 77% (SEM +/- 8%) of the initial CFU-GM remained.