John W. Theus

Viability and engraftment of hematopoietic progenitor cells after long-term cryopreservation: effect of diagnosis and percentage dimethyl sulfoxide concentration

BACKGROUND AIMS We carried out a retrospective analysis of viability by diagnosis and dimethyl sulfoxide (DMSO) concentration in patients who had undergone autologous transplants using hematopoietic progenitor cells (HPC) after long-term storage (up to 17.8 years). METHODS Viability was tested using flow cytometry for HPC that were harvested and preserved using a controlled rate freezer and 5% or 10% DMSO with human serum albumin, then stored in liquid nitrogen. Data from 262 samples were analyzed (249 myeloma patients and 13 other diagnoses): 100 consecutively thawed samples with a storage time of <1 year (all 10% DMSO), 50 consecutive samples stored for 1-4.9 years (10% DMSO), 50 samples stored for 5-9 years (5% DMSO) and all samples stored and used for transplant after >9 years (60 samples, 5% DMSO; two samples, 10% DMSO). RESULTS No statistically significant difference in viability between the 5% DMSO and 10% DMSO groups was observed (P = 0.08), so the 1-4.9 years and 5-9 years were combined and the three groups (<1 year, 1-9 years and >9 years) were compared using an anova test. There was no difference in viability based on cryostorage period (P = 0.23) or between myeloma and other diagnoses (P = 0.45). No difference was seen in time to White blood cell (WBC) engraftment (P = 0.10) or to platelet engraftment between groups (P = 0.52). CONCLUSIONS These data suggest that long-term storage in 5% DMSO and human serum albumin is safe.

A comparison of washed and volume-reduced platelets with respect to platelet activation, aggregation, and plasma protein removal.

BACKGROUND: Washed or volume‐reduced platelets (PLTs) are occasionally requested for patients with a history of allergic or anaphylactic transfusion reactions. However, conclusive data are not available as to which method is more suitable.

Red blood cell transfusion: Impact of an education program and a clinical guideline on transfusion practice

BACKGROUND Red blood cell (RBC) transfusion guidelines have been developed by professional societies. These guidelines recommend a restrictive RBC transfusion practice for most clinical populations. Despite the consistency of guidelines and limited evidence for RBC transfusion efficacy, there is variability in RBC transfusion practice. METHODS A program was initiated in a tertiary medical center to align RBC transfusion practice with best-practice RBC transfusion guidelines. The program included an educational program, followed after 6 months by RBC transfusion decision support that included the approval of a best-practice RBC transfusion guideline by the hospital medical board and an RBC transfusion order form that included the guideline recommendations. RBC transfusion practice was followed over an 18-month period and compared with transfusion practice over the prior 18 months. The primary outcome variables were adult inpatient RBC units transfused, RBC units per admission, and RBC units per 100 patient-days. RESULTS The mean RBC units transfused decreased with initiation of each component of the program: from 923 ± 68 units to 852 ± 40 (P = 0.025) with education and further to 690 ± 52 (P < 0.0001) with the RBC transfusion decision support. Similarly, RBC transfusions per 100 patient-days fell from 10.56 ± 0.80 to 9.69 ± 0.49 (P = 0.02) and to 7.68 ± 0.63 (P = 0.0001) during the 3 time periods. CONCLUSION An education program coupled with institutional adoption of a best-practice RBC transfusion guideline and RBC transfusion order set resulted in a reduction in total RBC units transfused.

Comparison of Bromcresol Purple and Capillary Protein Electrophoresis for Quantification of Serum Albumin in Multiple Myeloma

According to the International Staging System (1), serum albumin concentration is a key factor in determining patient prognosis in multiple myeloma. A recent study published in Clinical Chemistry described significant discordance between bromcresol green (BCG)1 albumin dye and agarose gel serum protein electrophoresis (SPEP) for albumin concentration determination in patients with high concentrations of monoclonal (M)- protein (2). Also available, however, are commonly used alternative methods for albumin determination, such as bromcresol purple (BCP) albumin dye and capillary zone SPEP (3). To examine whether the previously observed comparative discordance of measured albumin concentration is limited only to the BCG dye and agarose gel SPEP methods of albumin determination (2), we retrospectively compared measured albumin concentrations as determined by BCP albumin dye and capillary zone SPEP in a large cohort of multiple myeloma patients. The ratio of measured BCP albumin to SPEP albumin concentrations in patients with a previously identified monoclonal protein was retrospectively calculated for 579 specimens submitted to the University of Arkansas Immunology laboratory for routine serum protein electrophoresis by capillary zone SPEP. All specimens had concurrent albumin determination by a BCP albumin dye method. These consisted of 355 IgG, 122 IgA, 29 IgM, 13 IgD, and 60 free-light-chain myeloma patient specimens. …

A correlation between growth rate, apoptosis, and tumor necrosis factor‐α in umbilical cord blood cells infected with two strains of Mycobacterium tuberculosis

BACKGROUND: Using umbilical cord blood (UCB) cells, it was demonstrated that three virulent isolates, two highly transmissible clinical isolates and the virulent laboratory strain, demonstrated rapid growth in the UCB cells, which was significantly faster than the growth rate observed for a unique isolate. There was also a significant increase in the amount of tumor necrosis factor (TNF)‐α elicited from the UCB cells after infection with the unique isolate compared to the hypervirulent isolates. This study investigated whether neutralization or addition of TNF‐α within this system would alter growth rates and apoptosis.

UC blood infection with clinical strains of Mycobacterium tuberculosis: a novel model

BACKGROUND Use of unrelated cord blood transplantation (UCBT) is increasing, yet high rates of mortality secondary to infection remain a problem. We investigated the utility of using umbilical cord blood (UCB) as a model to study a naive cell population challenged by Mycobacterium tuberculosis. METHODS Mononuclear cells were isolated from nine UCB samples and infected with each of four distinct strains of M. tuberculosis. The isolates used were two highly transmissible clinical strains, the virulent laboratory strain H37Rv and a unique strain isolated from only one case (i.e. non-virulent). CFU were assessed at 3 h post-infection (day 0) and at day 7 to generate growth curves. Viability of the mononuclear cells was assessed prior to infection, 3 h post-infection and at days 3, 5 and 7 post-infection. IFN-gamma and TNF-alpha levels were determined at 24 h post-infection. RESULTS All three of the virulent strains demonstrated rapid growth in UCB cells that was significantly faster than the growth rate observed for the non-virulent unique isolate. There was no significant decrease in UCB cell viability after the 7-day incubation period regardless of infecting isolate. UCB cells secreted IFN-gamma in response to infection, with no significant difference related to infection with different isolates. However, there was a significant increase in the amount of TNF-alpha elicited following infection with the non-virulent isolate compared with the virulent isolates. DISCUSSION These results show that UCB can be used as a model to study infection, hopefully leading to new therapies for neonates and UCBT recipients.