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In March 2020, there were no treatment options for COVID-19. Passive immune therapy including anti-SARS-CoV-2 hyperimmune globulin (hIVIG) was a logical candidate for COVID-19 therapeutic trials, given past success treating emerging pathogens with endogenous neutralizing antibodies. We established a plasma collection protocol for persons recovered from COVID-19. To speed recruitment in the first U.S. hotspot, Seattle, Washington, federal and state public health agencies collaborated with Bloodworks Northwest to collect convalescent plasma (CP) for manufacturing hIVIG. During March–December 2020, we identified and recruited prospective CP donors via letters to persons recovered from COVID-19 with laboratory-confirmed SARS-CoV-2 infection. Prospective donors were pre-screened and administered a medical history survey. Anti-SARS-CoV-2 neutralizing antibody (NAb) titers were classified as qualifying (≥1:80) or non-qualifying (

Two case series examining the impact of convalescent plasma on patients with COVID-19 suggest some clinical benefit from early administration and modest impact on parameters of inflammation. Further assessment of the impact of this intervention awaits controlled clinical trials.

Background Platelet inventory constraints can result in minor ABO incompatibility and possible hemolysis. The aims of this study were to determine the reduction of isoagglutinin in titers of platelets stored in additive solution (PAS) and compare its safety, efficiency, and cost-effectiveness with full-volume and plasma-reduced platelets. Study design and methods Isoagglutinin titers were performed in paired whole blood donor samples and apheresis platelets collected in PAS (PAS-PLT) aliquot samples by the tube method. Results A total of 149 pairs of donor/platelet samples were tested: 75 group O, 59 group A, and 15 group B. For group O donor samples, the median anti-A IgG and IgM were 64 and 16, respectively, and the median anti-B IgG and IgM were 64 and 16, respectively. For group O PAS-PLT samples the mean anti-A IgG and IgM, and anti-B IgG and IgM were 32 and 8, and 16 and 8, respectively. For group A donor samples, the mean anti-B IgG and IgM was 8 in both cases; and both titers decreased to 2 in PAS-PLT. For group B donor samples, mean anti-A IgG and IgM was 16 in both cases; and both titers decreased to 4 in PAS-PLT. PAS-PLT demonstrated a net reduction in cost and improved efficiency when compared to plasma reduction. The use of PAS-PLT resulted in a 40% reduction of allergic transfusion reactions. Conclusion The use of PAS decreases plasma isoagglutinin titers, transfusion reactions, and is cost-effective when compared to routine plasma reduction as a strategy to mitigate hemolysis risk from minor incompatible platelet transfusion.

Platelet inventory constraints can result in minor ABO incompatibility and possible hemolysis. The aims of this study were to determine the reduction of isoagglutinin in titers of platelets stored in additive solution (PAS) and compare its safety, efficiency, and cost-effectiveness with full-volume and plasma-reduced platelets.Isoagglutinin titers were performed in paired whole blood donor samples and apheresis platelets collected in PAS (PAS-PLT) aliquot samples by the tube method.A total of 149 pairs of donor/platelet samples were tested: 75 group O, 59 group A, and 15 group B. For group O donor samples, the median anti-A IgG and IgM were 64 and 16, respectively, and the median anti-B IgG and IgM were 64 and 16, respectively. For group O PAS-PLT samples the mean anti-A IgG and IgM, and anti-B IgG and IgM were 32 and 8, and 16 and 8, respectively. For group A donor samples, the mean anti-B IgG and IgM was 8 in both cases; and both titers decreased to 2 in PAS-PLT. For group B donor samples, mean anti-A IgG and IgM was 16 in both cases; and both titers decreased to 4 in PAS-PLT. PAS-PLT demonstrated a net reduction in cost and improved efficiency when compared to plasma reduction. The use of PAS-PLT resulted in a 40% reduction of allergic transfusion reactions.The use of PAS decreases plasma isoagglutinin titers, transfusion reactions, and is cost-effective when compared to routine plasma reduction as a strategy to mitigate hemolysis risk from minor incompatible platelet transfusion.

The cell source for hematopoietic cell transplantation (HCT) can be autologous, related allogeneic, or unrelated allogeneic. Autologous cell therapy (CT) and related allogeneic CD34+ cell products are often collected and processed in the same facility as the transplant center. This is not the case, however, for unrelated donor cord blood, peripheral blood, or bone marrow transplantation that are facilitated by registries—such as the National Marrow Donor Program (NMDP) and the World Marrow Donor Association (WMDA)—that are involved in the public exchange of these allogeneic CT products worldwide. Non-cryopreserved and cryopreserved allogeneic CT products are, therefore, frequently transferred on public roads as well as on aircrafts to geographically distant facilities. Since the infusion is usually scheduled within 48–72 h after collection (for non-cryopreserved CT products) or after receipt at the transplant center (for cryopreserved CT products), this process mandates complex coordination by highly trained personnel. The process of transferring CT products is tightly regulated by the Food and Drug Administration (FDA), Department of Transportation (DOT), International Air Transport Association (IATA), International Civil Aviation Organization (ICAO), AABB (formerly known as the American Association of Blood Banks), and Foundation for the Accreditation of Cellular Therapy (FACT). The regulations that must be followed depend on the hazardous material classification of the biological and cryogenic substances in the transfer container. The requirements for continuous temperature monitoring and the procedures for packaging, labeling, and documentation are all designed to maintain the integrity of the CT product while protecting the health and safety of personnel involved in the transfer process. It is essential to clearly document the chain of custody as the CT product is transferred from the cosigner (transfer facility) to the consignee (receiving facility) via the courier. Upon receipt, trained personnel at the receiving facility should promptly follow the instructions for opening the container and inspecting the CT product as well as make a decision to accept, reject, or quarantine the CT product.

Hakha Chin—a Tibeto-Burman language spoken in Chin State in western Myanmar—is reported to have phonemic length distinctions in its monophthongs (Melnik, 1997; Peterson, 2003; Maddieson, 2004) (e.g., [ii] vs. [i], [aa] vs. [a]). These pairs are also sometimes represented with tense/lax vowels (e.g., [i] vs. [ɪ], [a] vs. [ə]) (Peterson, 2017). Indeed, the results of a perception study conducted by Mortenson and Van Bik (2002) found variable response patterns in different dialect groups. Listeners from Hakha, the city after which the language is named, relied heavily on spectral cues related to quality to distinguish between long and short vowels, while listeners from the town of Thanthlang relied mainly on durational cues to perform the same task. In the current study, we present the first instrumental acoustic analysis of Hakha Chin vowel quality in order to aid in elucidating the nature of the contrast. Data from three college-aged native speakers (two male) suggest that neither duration nor quality play key roles in distinguishing between the so-called long and short vowels of Hakha Chin. Rather, as originally suggested by Maddieson (2004), the length associated with long vowels is mainly realized through lengthening of sonorant codas.

Background. Many cancers, including melanoma, exclusively express constitutive proteasomes (cPs) and are unable to express immunoproteasomes (iPs). In contrast, mature DCs used for immunotherapy exclusively express iPs. Since proteasomes generate peptides presented by HLA class I molecules, we hypothesized that mature melanoma antigen–loaded DCs engineered to process antigens through cPs would be superior inducers of antimelanoma immunity in vivo. Methods. Subjects with metastatic melanoma were vaccinated with mature DCs transfected with RNAs encoding melanoma antigens MART1, MAGE-3, gp100, and tyrosinase. These DCs were derived from monocytes that were untransfected (Arm A; n = 4), transfected with control siRNA (Arm B; n = 3), or transfected with siRNAs targeting the 3 inducible iP subunits (Arm C; n = 5). Results. Vaccination stimulated antigen-specific T cell responses in all subjects, which peaked after 3–4 vaccinations, but remained elevated in Arm C subjects. Also in Arm C, circulating melanoma cell levels (as detected by quantitative PCR) fell, and T cell lytic activity against autologous melanoma was induced. In HLA-A2+ subjects, CD8+ T cells that bound tetramers loaded with cP-derived melanoma antigenic peptides were found in the peripheral blood only in Arm C subjects. Of 2 subjects with active disease (both in Arm C), one had a partial clinical response, while the other, who exhibited diffuse dermal and soft tissue metastases, had a complete response. Conclusion. These results suggest that the efficacy of melanoma DC–based immunotherapy is enhanced when tumor antigen–loaded DCs used for vaccination express cPs. Trial registration. Clinicaltrials.gov {"type":"clinical-trial","attrs":{"text":"NCT00672542","term_id":"NCT00672542"}}NCT00672542. Funding. Duke Clinical Research Institute/Duke Translational Medicine Institute, Duke Melanoma Consortium, and Duke University Department of Surgery.

An acronym, a button, a script card, and a lot of fun are all an educator needs to turn dreaded education into a great learning experience for nurses as well as the patients. A multidisciplinary team pilot tested a new learning approach on a cardiac step-down unit. The goal was to educate both nurses and patients about the American Heart Association's Get-With-the-Guidelines Program for Coronary Artery Disease. The educational strategies were successful, and data revealed an increase in nursing knowledge of core measures. After a two-phase study, the program was eventually rolled out hospital-wide. The study results show that nurses' compliance with and knowledge of the AHA core measures increased as a direct result of the program.

Cancer vaccines have now demonstrated clinical efficacy, but immune modulatory mechanisms that prevent autoimmunity limit their effectiveness. Systemic administration of mAbs targeting the immune modulatory receptors CTLA-4 and glucocorticoid-induced TNFR-related protein (GITR) on Treg and effector T cells augments anti-tumor immunity both experimentally and clinically, but can induce life-threatening autoimmunity. We hypothesized that local delivery of anti-CTLA-4 and anti-GITR mAbs to the sites where T cells and tumor antigen-loaded DC vaccines interact would enhance the induction of anti-tumor immunity while avoiding autoimmunity. To achieve this goal, DCs transfected with mRNA encoding the H and L chains of anti-mouse CTLA-4 and GITR mAbs were co-administered with tumor antigen mRNA-transfected DCs. We observed enhanced induction of anti-tumor immunity and significantly improved survival in melanoma-bearing mice, without signs of autoimmunity. Using in vitro assays with human DCs, we demonstrated that DCs transfected with mRNA encoding a humanized anti-CTLA-4 mAb and mRNA encoding a soluble human GITR-L fusion protein enhance the induction of anti-tumor CTLs in response to DCs transfected with mRNAs encoding either melanoma or breast cancer antigens. Based on these results, this approach of using local delivery of immune modulators to enhance vaccine-induced immunity is currently being evaluated in a phase I clinical cancer immunotherapy trial.

Objectives: The safety and efficacy of direct intramuscular injections of aldehyde dehydrogenase bright (ALDH br ) cells isolated from autologous bone marrow mononuclear cells (ABMMNCs) and ABMMNCs were studied in patients with critical limb ischemia (CLI) who were not eligible for percutaneous or surgical revascularization. Background: Many CLI patients are not candidates for current revascularization procedures, and amputation rates are high in these patients. Cell therapy may be a viable option for CLI patients. Methods: Safety was the primary objective and was evaluated by occurrence of adverse events. Efficacy, the secondary objective, was evaluated by assessment of Rutherford category, anklebrachial index (ABI), transcutaneous partial pressure of oxygen (TcPO2), quality of life, and pain. Results: ALDH br cells and ABMMNCs were successfully administered to all patients. No therapy-related serious adverse events occurred. Patients treated with ALDH br cells (n 5 11) showed significant improvements in Rutherford category from baseline to 12 weeks (mean, 4.09 6 0.30 to 3.46 6 1.04; P 5 0.05) and in ABI at 6 (mean, 0.22 6 0.19 to 0.30 6 0.24; P 5 0.02), and 12 weeks (mean, 0.36 6 0.18; P 5 0.03) compared with baseline. Patients in the ABMMNC group (n 5 10) showed no significant improvements at 6 or 12 weeks in Rutherford category but did show improvement in ABI from baseline to 12 weeks (0.38 6 0.06 to 0.52 6 0.16; P 5 0.03). No significant changes from baseline were noted in ischemic ulcer grade or TcPO2 in either group. Conclusions: Administration of autologous ALDH br cells appears to be safe and warrants further study in patients with CLI. V C 2011 Wiley Periodicals, Inc.

BACKGROUND: Cryopreservation of hematopoietic progenitor cells using liquid nitrogen and controlled-rate freezing requires complex equipment and highly trained staff and is expensive. We compared the liquid nitrogen method with methods using a combination of dimethyl sulfoxide (DMSO) and hydroxyethyl starch (HES) for cryopreservation followed by storage in mechanical freezers. STUDY DESIGN AND METHODS: Peripheral blood stem cells (PBSCs) were collected from normal donors by apheresis and allocated to one of four preservation and storage conditions: 1) 10% DMSO with freezing in liquid nitrogen and storage in liquid nitrogen, 2) 5% DMSO and 6% HES with freezing and storage in a −80°C mechanical freezer, 3) 5% DMSO and 6% HES with freezing in a −80°C mechanical freezer and storage in a −135°C mechanical freezer, or 4) 5% DMSO and 6% HES with freezing and storage both in a 135°C mechanical freezer. Cells were stored for 5 years during which total nucleated cells (TNCs), cell viability, CD34+ cell content, and colony-forming unit–granulocyte-macrophage content were determined. RESULTS: There were some significant differences in the variables measured during freezing and the 5 years of storage compared to the values before freezing and storage; however, these differences were not consistent and do not favor one protocol over the others. Samples stored for 24 hours before cryopreservation showed a significant decrease in TNCs, but no other significant changes during the 5 years. CONCLUSION: In vitro measurements indicate that PBSCs can be successfully frozen and stored using a combination of DMSO and HES providing smaller amounts of DMSO and allowing simplified freezing and storage conditions.

Aim: Peripheral blood-derived endothelial cells (pBD-ECs) are an attractive tool for cell therapies and tissue engineering, but have been limited by their low isolation yield. We increase pBD-EC yield via administration of the chemokine receptor type 4 antagonist AMD3100, as well as via a diluted whole blood incubation (DWBI). Materials & Methods: Porcine pBD-ECs were isolated using AMD3100 and DWBI and tested for EC markers, acetylated LDL uptake, growth kinetics, metabolic activity, flow-mediated nitric oxide production and seeded onto titanium tubes implanted into vessels of pigs. Results: DWBI increased the yield of porcine pBD-ECs 6.6-fold, and AMD3100 increased the yield 4.5-fold. AMD3100-mobilized ECs were phenotypically indistinguishable from nonmobilized ECs. In porcine implants, the cells expressed endothelial nitric oxide synthase, reduced thrombin-antithrombin complex systemically and prevented thrombosis. Conclusion: Administration of AMD3100 and the DWBI method both increase pBD-EC yield.

Objective.To characterize red eye reactions occurring within 24 hours after receipt of units of leukocyte-reduced red blood cells, determine their etiology, and investigate their potential link to transfusion.Methods.We conducted a survey of transfusion facilities nationwide to determine the scope and magnitude of the reactions; performed case-control and cohort studies among transfused patients at the facility where most reactions occurred; and performed animal experiments, using cellulose acetate derivatives extracted from leukocyte-reduction filters and filter precursors, to reproduce reactions.Results.From January 1, 1997, through January 15, 1998, we identified 159 reactions in 117 patients from 17 states. Reactions were characterized by conjunctival erythema or hemorrhage (in 100% of patients), eye pain (in 62%), photophobia (in 46%), and decreased visual acuity (in 32%). Symptom onset occurred 1-24 hours after initiation of transfusion and resolved within a median of 5 days. Reactions were associated with transfusion sessions that included units of red blood cells filtered with a specific brand of filter, the LeukoNet filter (HemaSure) (odds ratio, 100.4; P< .001). There was a dose-response relationship between the number of LeukoNet-filtered units transfused and the attack rate for reactions, ranging from 0.8% among sessions in which 1 unit was transfused to 27.3% among sessions in which 3 or more units were transfused (P< .001). A similar ocular syndrome was elicited in rabbits injected with cellulose acetate derivatives extracted from unused LeukoNet filters or filter precursors. No reactions were reported after LeukoNet filters were withdrawn from the market.Conclusions.This transfusion-associated red eye syndrome was linked to a specific brand of leukocyte-reduction filter and likely resulted from cellulose acetate derivatives leached from the filter membrane.

Banked, unrelated, partially HLA-matched, umbilical cord blood is an alternative stem cell source for patients in need of transplantation therapy who lack traditionally matched donors. A presumed advantage of cord blood is the ability to increase recruitment of donors of minority ethnic backgrounds. The American Red Cross Cord Blood Program was established in 1999 with 6 banks and 10 collection sites throughout the country. Cord blood donors self-report racial designations on questionnaires, and donor race was collected from each site. Postprocessing nucleated cell counts and CD34(+) counts were obtained on the cord blood units, and results from each racial group (white, black, Asian, Hispanic, and Native American) were compared in the natural logarithmic scale by using analysis of variance. A total of 18878 donors consented: 64% white, 16% black, 12% Hispanic, 4% Asian, 1% Native American, and 3% other. The Detroit area consented the highest percentage of black donors (87%), San Diego consented the highest percentage of Hispanic donors (59%), and Oakland consented the highest percentage of Asian donors (15%). Seven thousand eight hundred sixty-six cord blood units have been banked for transplantation. The mean preprocessing nucleated cell count was 1220 x 10(6) (range, 327-7300 x 10(6)). There was no difference among racial groups when controlled for site (P =.395). The mean CD34(+) count was 3.28 x 10(6). Blacks had a significantly lower CD34(+) count than the other racial/ethnic groups in the Midwest, Northwest, and North Carolina collection sites. A racially diverse cord blood bank can be achieved. Nucleated cell counts were similar among the different racial/ethnic groups. CD34(+) counts were lower for blacks in some collection sites.

It has been suggested that the work environment of the United States Congress bears similarity to a fixed-interval reinforcement schedule. Consistent with this notion, Weisberg and Waldrop (1972) described a positively accelerating pattern in annual congressional bill production (selected years from 1947 to 1968) that is reminiscent of the scalloped response pattern often attributed to fixed-interval schedules, but their analysis is now dated and does not bear on the functional relations that might yield scalloping. The present study described annual congressional bill production over a period of 52 years and empirically evaluated predictions derived from four hypotheses about the mechanisms that underlie scalloping. Scalloping occurred reliably in every year. The data supported several predictions about congressional productivity based on fixed-interval schedule performance, but did not consistently support any of three alternative accounts. These findings argue for the external validity of schedule-controlled operant behavior as measured in the laboratory. The present analysis also illustrates a largely overlooked role for applied behavior analysis: that of shedding light on the functional properties of behavior in uncontrolled settings of considerable interest to the public.

SUMMARy While health care providers, governmental agencies and public interest groups in the United States show concern about blood safety and transfusion reactions, there is no unified national reporting system. This lack of unity derives largely from the structure (or lack thereof) of the health care system in the U.S. The hazards of transfusion are described by a patchwork of regulations, reports and studies, many of them voluntary. Even without a unified system, the United States does have examples of reports that cover the usual elements of a national reporting or monitoring system. Most of the elements of a national system have been undertaken by cooperative groups, such as the BaCon Study Group, or by smaller but very informative public studies, such as the State of New york study of transfusion errors. The collection of very basic information-how many units of blood are collected and where and what components are made-is privately handled and incompletely funded. The current reporting practices lack pro-active approaches in dealing with emerging infectious diseases capable of being spread by blood and tissue. The West Nile Virus infections from blood and tissue that occurred last summer are examples of full investigations that were triggered by patient deaths rather than early reports.

The hematopoietic progenitor cells present in the umbilical cord (HPC, Cord Blood) have the potential to engraft and differentiate in the bone marrow with subsequent trilineage hematopoiesis. The collection, processing and storage of HPC, Cord Blood should preserve the cell number and characteristics to protect their hematopoietic potential. Federal regulations establish guidelines for HPC, Cord Blood manufacturing and testing to ensure the safety, purity, potency and identity of the product. Accrediting organizations for cord blood banks, including the AABB and the Foundation for Accreditation of Cellular Therapy (FACT), provide a series of guidelines for developing safe processes and procedures in cellular therapy activities. Additionally, clinical data based on outcome analysis provide evidence to help in the process of unit selection for a given patient, based on compatibility, cell dose and potency of the product.

BACKGROUND: Bacterial contamination of blood components can result in transfusion-transmitted infection, but the risk is not established. STUDY DESIGN AND METHODS: Suspected cases of transfusion-transmitted bacteremia were reported to the CDC by participating blood collection facilities and transfusion services affiliated with the American Red Cross, AABB, or Department of Defense blood programs from 1998 through 2000. A case was defined as any transfusion reaction meeting clinical criteria in which the same organism species was cultured from a blood component and from recipient blood, with the organism pair confirmed as identical by molecular typing. RESULTS: There were 34 cases and 9 deaths. The rate of transfusion-transmitted bacteremia (in events/million units) was 9.98 for single-donor platelets, 10.64 for pooled platelets, and 0.21 for RBC units; for fatal reactions, the rates were 1.94, 2.22, and 0.13, respectively. Patients at greatest risk for death received components containing gram-negative organisms (OR, 7.5; 95% CI, 1.3-64.2; p = 0.009). CONCLUSION: Bacterial contamination of blood is an important cause of transfusion-transmitted infection; infection risk from platelet transfusion is higher compared with that from RBCs, and, overall, the risk of infection from bacterial contamination now may exceed that from viral agents. Recipients of components containing gram-negative organisms are at highest risk for transfusion-related death. The results of this study may help direct efforts to improve transfusion-related patient safety.

BACKGROUND: The transfusion of blood components contaminated with bacteria may have serious clinical consequences, but few data are available on the incidence of these events. A national effort to assess the frequency of blood component bacterial contamination associated with transfusion reaction (the BaCon Study) was initiated to better estimate their occurrence. STUDY DESIGN AND METHODS: Standard reporting criteria, data collection forms, and a standardized reporting protocol were developed in collaboration with the American Red Cross, AABB, and the Department of Defense. Episodes reported to the BaCon Study were compared with those reported to the FDA's national reporting systems to estimate the extent to which all serious reactions associated with bacterial contamination were captured. RESULTS: During the first 2 years, 38 episodes meeting study criteria were reported; 21 were laboratory-confirmed. The estimated proportion of episodes reported to the BaCon Study (i.e., completeness of coverage) was lower than that reported to the FDA during the same period (0.33 vs. 0.68), but the positive predictive value was higher (0.66 vs.0.28). CONCLUSION: Despite the complexity of obtaining reports from a large number of United States hospitals and transfusion centers, the feasibility and usefulness of the BaCon Study were shown. This study was the only national study in the United States to monitor adverse clinical events associated with bacterial contamination of blood components. By building on hospital-based reporting of transfusion-related adverse events, the BaCon Study serves as a model for the study of other complications associated with blood and blood components.

The purpose of this in vitro study was to determine whether the Gemini PC-2TX infusion pump could safely deliver peripheral stem cells (PSC) for an autologous PSC transplant. For purposes of hypothesis testing, it was assumed that there would be no significant difference in CD34+ cell counts and colony-forming units-granulocyte, macrophage (CFU-GM) when the PSCs were administered by an IMED PC-2TX infusion pump as opposed to an intravenous push method. The American Red Cross collected 50-ml samples of PSCs from four donors by apheresis. These cells were tested for CD34+ using flow cytometry and for functional progenitor cells using a CFU-GM assay. The cells were cryopreserved after testing. For our study, samples were tested simultaneously at a single facility. Each sample was individually thawed and a baseline thaw sample collected; 10 ml of the donor specimen was pushed through a syringe into a specimen container (intravenous push sample). The remainder of the specimen was infused through the IMED Gemini PC-2TX pump into a specimen container (intravenous pump sample). All samples were assayed for CD34+ cell counts and CFU-GM. Data analyses were conducted using the t-test for paired samples, with values of P < 0.05 considered significant. Results failed to demonstrate a statistically significant difference between the CD34+ or CFU-GM results of the intravenous push and intravenous pump specimens. Additionally, we failed to find a statistically significant difference when we compared the intravenous push and the intravenous pump specimens with the baseline thaw sample. The results of this study support the hypothesis that the Gemini PC-2TX infusion pump can safely deliver PSCs for the purposes of stem cell transplantation.

Endothelial cells (ECs) isolated from endothelial progenitor cells in blood have great potential as a therapeutic tool to promote vasculogenesis and angiogenesis and treat cardiovascular diseases. However, current methods to isolate ECs are limited by a low yield with few colonies appearing during isolation. In order to utilize blood-derived ECs for therapeutic applications, a simple method is needed that can produce a high yield of ECs from small volumes of blood without the addition of animal-derived products. For the first time, we show that human endothelial cells can be isolated without the prior separation of blood components through the technique of diluted whole blood incubation (DWBI) utilizing commercially available human serum. We isolated ECs from small volumes of blood (~ 10 ml) via DWBI and characterized them with flow cytometry, immunohistochemistry, and uptake of DiI-labeled acetylated low density lipoprotein (DiI-Ac-LDL). These ECs are functional as demonstrated by their ability to form tubular networks in Matrigel, adhere and align with flow under physiological fluid shear stress, and produce increased nitric oxide under fluid flow. An average of 7.0 ± 2.5 EC colonies that passed all functional tests described above were obtained per 10 ml of blood as compared to only 0.3 ± 0.1 colonies with the traditional method based on density centrifugation. The time until first colony appearance was 8.3 ± 1.2 days for ECs isolated with the DWBI method and 12 ± 1.4 days for ECs isolated with the traditional isolation method. A simplified method, such as DWBI, in combination with advances in isolation yield could enable the use of blood-derived ECs in clinical practice.

A comparison of the responses to recombinant human erythropoietin in normal and uremic subjects. The erythropoietic response to graded doses of recombinant human erythropoietin (epoetin alfa) was assessed in 24 hemodialysis patients by quantitative ferrokinetic studies, and measurement of the reticulocyte count and plasma levels of transferrin receptor protein. These responses were compared to those of 22 normal subjects. Epoetin alfa was given intravenously at 15, 50 or 150 U/kg every other day for four injections. Three patients with chronic renal failure were restudied after renal function was restored following renal transplantation. The results of these three different measurements of erythroid function showed that the acute response to recombinant human erythropoietin was similar in normal subjects and patients with renal failure. We conclude that chronic uremia does not alter the responsiveness to erythropoietin in vivo.

BACKGROUND Recent evidence demonstrates an association between duration of storage of red blood cells (RBC) and morbidity and mortality after cardiac surgery. We studied the feasibility of two different schemes for categorizing and randomizing age of RBC units transfused in cardiac surgical patients. STUDY DESIGN AND METHODS In Phase 1, 20 subjects were randomly assigned to standard of care (SOC) versus no RBCs with more than 21 days' storage duration. In Phase 2, 23 subjects were randomized to RBCs of 7 +/- 4 versus 21 +/- 4 days' storage duration. The age of study RBC units was masked. RESULTS In Phase 1, no patients received RBCs 31 days or older in SOC, and there was overlap in storage age shared in both arms so the predefined feasibility criteria were not met. In Phase 2, it was feasible to deliver specified age RBCs to the 7-day arm (achieved in 100% of subjects), but feasibility was not demonstrated for the 21-day arm (only 50% of subjects transfused with target age RBCs). Significant differences, however, were observed between the 7 +/- 4- and 21 +/- 4-day arms with respect to age of all RBC units (6 +/- 2 vs. 18 +/- 7, p = 0.0002) and maximum age (7 +/- 2 vs. 20 +/- 7, p < 0.0001). CONCLUSION Given the current storage age distribution of available RBC inventory, use of a SOC arm in future studies is unlikely to result in a large exposure to "old" blood. It is feasible to randomize patients to "younger" RBCs (3-11 days) but design strategies are needed to provide "intermediate-aged" or "old" blood as a comparator.

BACKGROUND : The relative nucleated cell count of umbilical cord blood (CB) correlates with improved engraftment and survival. This study compares two collection methods to assess CB content, including cell numbers. STUDY DESIGN AND METHODS : The Massachusetts CB bank used trained obstetricians and midwives to collect CB in utero before the delivery of the placenta. The banks in California, Ohio, Oregon, and Minnesota used trained American Red Cross (ARC) personnel who collected CB ex utero after the delivery of the placenta. All banks processed CB by RBC sedimentation and volume reduction. RESULTS : The volume and total nucleated cell count of collected CB before processing, as well as after processing CFU-GM and CD34+ cells, showed no advantage of either method. In utero collections resulted in more rejections of collected units (due to labeling problems, bacterial contamination, clotting, and delay between collection and processing) than ex utero collections. There were fewer medical exclusions after in utero collection. CONCLUSION : CB can be collected successfully using either the in utero or ex utero methods; both methods produce comparable nucleated cell, MNC, CD34+, and CFU-GM numbers. Bacterial contamination, low volume, clotting, and delay until processing are generally higher with in utero collection.

This project is aimed at developing a cost-effective working environment for the transfusion medicine specialists of American Red Cross (ARC). In this project we are developing a multimedia-based consultation environment that uses Internet and teleconferencing to increase the quality of services and to replace currently used 800 telephone lines. Through the use of Internet/LAN/ISDN the physicians can share information and references while they discuss patient cases. A multimedia interface allows the physician to access data from the office and from the house. This paper discusses the approach, current status of the project and future plans to extend the approach to other areas of medicine.© (1995) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.

Background: Dendritic cells (DC) must be matured to maximally stimulate antigen-specific immune responses and avoid the induction of tolerance. During maturation, the proteasome, the multi-subunit complex that generates peptides presented in the context of HLA class I, is altered to exclusively consist of immunoproteasomes (iPs). Compared to the constitutive proteasome (cP), the iP has altered protease activities and generates a different repertoire of peptide epitopes. Vaccination with mature tumor-antigen loaded DC will thus stimulate immunity against iP-generated peptides. Since melanomas, and other cancers, are frequently defective in inducible iP expression, this anti-tumor immune response may be miss-targeted and not recognize melanomas presenting cP-derived peptides. We hypothesized that mature melanoma antigen-loaded DC expressing the cP, rather than the iP, would be superior inducers of anti-melanoma immunity in subjects with metastatic melanoma. Methods: 12 subjects with metastatic melanoma were immunized with 6 weekly intradermal injections of 10 million mature autologous monocyte-derived DC transfected with RNA encoding full-length MART-1, MAGE-3, gp100 and tyrosinase. The monocytes from which the DC were generated were either untransfected (Study Arm A, n=4), transfected with control siRNA (Study Arm B, n=3), or transfected with siRNAs targeting the 3 inducible iP subunits LMP-2, LMP-7 and MECL-1 (Study Arm C, n=5). Results: 10 subjects received a full course of vaccination, while only enough DC were available for 4 and 5 total vaccine doses in 2 subjects. All 12 subjects tolerated vaccination without toxicity. Vaccination stimulated melanoma antigen-specific CD4+ and CD8+ T cell responses detected in peripheral blood by IFN-γ ELISPOT in all subjects, which peaked after 3-4 vaccinations. These T cell responses decreased thereafter in subjects in Study Arms A and B, but remained elevated in subjects in Study Arm C. Melanoma cell levels in peripheral blood, as detected by qPCR, fell after vaccination in Study Arm C. Using autologous melanoma cells (and autologous normal skin fibroblasts as controls) derived from each subject, we found that vaccination in Study Arm C, but not A or B, induced peripheral T cells with lytic activity against autologous melanoma. Two subjects had active disease, both in Study Arm C. The one subject with diffuse extremity in-transit disease had a partial clinical response with transient liquefaction of dermal lesions. The other subject with diffuse dermal and soft tissue metastatic lesions had complete resolution of detectable metastasis by PET scanning. Conclusion: The efficacy of melanoma immunotherapy using antigen RNA-transfected mature DC is enhanced when DC express the cP rather than the iP. We feel that this novel approach warrants further clinical investigation. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5373. doi:1538-7445.AM2012-5373

The objective of this study was to quantitate the improvement in exercise capacity produced in anemic chronic hemodialysis (HD) patients after correction of their anemia with recombinant human erythropoietin (rHuEPO). The maximal exercise capacity and quadriceps strength of 19 anemic HD patients was tested before and after correction of the anemia with rHuEPO. A progressive work exercise protocol (PWET) on a cycle ergometer was used to compare measurements of maximal oxygen uptake (VO2max), maximal heart rate, and subjective assessment of fatigue during the test. Measurements of quadriceps strength were performed before the cycle ergometer studies. At baseline, all patients had reduced VO2max (15.3 +/- 5.4 mL/kg/min) and maximal exercise heart rates (138.5 +/- 23.9 beats/min). rHuEPO increased the mean hematocrit from 21.2% to 35%, and this was associated with a 17% increase (P less than 0.0005) in the VO2max. At any specified work load, rHuEPO treatment decreased heart rate, minute ventilation, and the subjective perception of fatigue. Both isometric and isokinetic measurements of quadriceps strength were improved following administration of rHuEPO. The maximal exercise heart rate was decreased in comparison to the baseline measurements (P less than 0.04), suggesting that in contrast to normal subjects, HD patients stop exercise before oxygen transport limitations are reached. In this unselected group of chronic HD patients, rHuEPO produced clinically significant improvements in both aerobic exercise capacity and isometric and isokinetic quadriceps strength. The improvement in aerobic capacity was substantially less than would have been expected from the correction of a comparable degree of anemia in non-HD patients. None of the 19 treated patients attained the exercise performance level predicted for a sedentary normal subject.

Umbilical cord blood (UCB) transplantation results in delayed myeloid and platelet engraftment, likely due to lower cell dosing with cord blood. One strategy to overcome this problem is to augment the graft with ex vivo expanded cells. Previous studies have shown large expansion of progenitor cells in vitro, but when infused, these cells had little to no effects on hematopoietic reconstitution in the recipient. We hypothesized that these cells may have terminally differentiated in culture, loosing the potential for further expansion in vivo. The ALDHbr fraction of non-erythroid, non-monocytic cells is enriched for stem and progenitor cells. We performed preclinical studies showing that ALDHbr cells cultured ex vivo in SCF, FLT-3 and IL-7 expanded several fold with survival of both differentiated and immature cells. We designed a clinical trial to test whether infusion of primed (but not expanded) ALDHbr cells would be safe and would impact engraftment and survival after unrelated donor UCB transplantation (UCBT). ALDHbr cells were isolated from the 20% fraction of cryopreserved cord blood units (CBU) 6 days before thei nfusion of donor cells. The cells were cultured in SCF/FLT3/IL7 for 5 days. Release criteria included TNC with viability, negative bacterial/fungal cuture taken 2 days before planned infusion and negative gram stain. After written informed consent, patients were assigned to receive sorted cells (n=3) or sorted and primed cells (n=8) augmenting a standard UCBT which was administered with the 80% fraction of the CBU. The study cells were given 4 hours after the standard infusion on day 0. The primary study endpoint was safety and secondary endpoints: engraftment, Graft versus Host Disease (GvHD) and overall survival. Thus far, 11 patients (55% male, 82% Caucasian, median age 1.26 years) with pediatric malignant (n=5) or non-malignant (n=6) diagnoses have been infused. The median precryopreservation cell dose of the CBU was 13.89×10e7/kg and median cell dose infused 9.53×10e7/kg (range 4.13–18.6). The cell yield post sort ranged from 50,000–100,000 ALDHbr cells. After priming, the TNC content remained stable. Cells were infused without clinical reactions. The cummulative incidence (CI) of neutrophil engraftment by day +42 was 90.9% (95% CI 78–100) and no patient experienced graft failure. The CI of platelet engraftment (50K by 100 days) was 79.5% (95% CI 63.6–95.5). The median day to neutrophil (ANC 500/uL) and platelet engraftment (50K) were 17 (range 12–33) and 50 (range 38–90). Median CD4 counts at 100 and 180 days were 123, and 257/uL, higher than those seen with standard transplants. Overall survival at latest followup (1–9 months) was 90.9% (95% CI 73.9–100%). One death occurred in a newborn with a metabolic disease from congenital pulmonary hypertension, unrelated to the study. Although the study population is small, these results compare favorably to a similar population of patients in the COBLT study where the CI of ANC engraftment by day 42 was 78% (p=0.001), platelet engraftment was 50% and overall survival was 57%(p=.2). We conclude that isolation and priming of ALDHbr cells is feasible, infusion is safe and effects on engraftment and survival appear promising.