Granulocyte transfusions: efficacy in treating fungal infections in neutropenic patients following bone marrow transplantation

Background: A retrospective study was conducted to evaluate the efficacy of granulocyte transfusions in neutropenic patients with fungal infections following bone marrow transplantation. Study Design and Methods: Systemic fungal infection was detected in 87 patients during the first 100 days following bone marrow transplantation; 50 received granulocytes in addition to appropriate antifungal agents. The median age was 17 years in the transfused patients (range, 1.5–57) and 35 years in the nontransfused patients (range, 0.8–50). Granulocyte transfusions were given on a daily to twice-daily basis. To evaluate their responses, patients were categorized by infection type (candidal [n = 38] vs. noncandidal [n = 49]) and site (fungemia alone [n = 30] vs. invasive infection [n = 57]). Resolution of infection was defined as the resolution of signs and symptoms and negative cultures and/or histopathology. Results: No benefit of granulocyte transfusions could be shown in the resolution of infection in patients with either invasive noncandidal infection (29% in the transfused patients vs. 23% in the nontransfused patients, p > 0.1) or candidal sepsis (56% vs. 50%, p > 0.1). Among patients with delayed marrow recovery, no difference was seen in the resolution of infection in the transfused (25.9%) and nontransfused (50%) patients (p > 0.1); nor was any difference between the transfused and nontransfused patients evident in the duration of febrile episode associated with the fungal infection. Granulocyte transfusions were well tolerated, with the only complications being fever in 12 patients (24%), chills in 10 (20%), and respiratory distress in 2 (4%). Despite attempts to stratify by infection type, invasiveness, and marrow recovery, it was not possible to show any benefit of granulocyte transfusions in this group. Conclusions: It is likely that only through a prospective randomized trial can the question of the efficacy of granulocyte transfusions in treating fungal infections be conclusively answered.     

Protection of human polymorphonuclear leukocyte function from the deleterious effects of isolation, irradiation, and storage by interferon-gamma and granulocyte-colony-stimulating factor

BACKGROUND: Fungal infections represent a difficult challenge to clinicians caring for neutropenic patients with hematologic malignancies, as antifungal therapy often has limited success in that setting. One promising yet problematic alternative approach is leukocyte transfusion. The isolation of polymorphonuclear leukocytes (PMNs) induces apoptosis and functional deterioration, and irradiation to prevent transfusion-associated graft-versus-host disease causes further functional deterioration. STUDY DESIGN AND METHODS: The ability of interferon-gamma and granulocyte-colony-stimulating factor (G-CSF), used both alone and in combination, to protect PMNs after 0 or 20 hours' storage in cell culture (as a model for function after transfusion) and irradiation with 0, 5, or 30 Gy was studied. RESULTS: Without cytokine treatment, 20-hour-old PMNs showed marked apoptosis, no appreciable chemotaxis, and no ability to kill Candida albicans. In contrast, cytokine treatment significantly reduced apoptosis and protected chemotaxis, C. albicans killing, and surface-receptor expression from both storage and irradiation. Although the majority of the benefit appeared to be due to G-CSF, consistent trends suggested better function of PMNs after combined treatment with interferon-gamma and G-CSF. CONCLUSION: Judicious use of cytokines may preserve PMN function. These findings have important implications for the transfusion of PMNs to cytopenic patients.     

Buffy coat platelets stored in apyrase, aprotinin, and ascorbic acid in a suspended bag: combined strategies for reducing platelet activation during storage

BACKGROUND: Platelet activation is an important factor impeding the clinical effectiveness of platelet transfusions. In this study, platelet concentrates (PCs) were prepared by a novel suspended-bag buffy coat technique that was followed by the addition of a mixture of platelet activation inhibitors to the storage bag. STUDY DESIGN AND METHODS: In vitro platelet function was evaluated in PCs prepared by the suspended-bag buffy coat technique and stored at 22 degrees C for 5 days in the presence of (n = 12) or absence (n = 12) of apyrase, ascorbic acid, and aprotinin (AAA). RESULTS: Platelets from AAA- incubated PCs demonstrated mean ATP levels 17 percent (p < 0.004), 13 percent (p < 0.02), and 22 percent (p < 0.003) higher than those measured in parallel control PCs on Days 1, 3, and 5, respectively. Similarly, on Days 3 and 5 of storage, respectively, 45-percent (p < 0.001) and 50-percent (p < 0.001) greater ADP-induced maximum aggregation was observed in AAA-incubated PCs than was seen in control preparations. AAA-incubated PCs demonstrated alpha-granule membrane protein-140 expression 92 percent (p < 0.01), 133 percent (p < 0.003), and 104 percent (p < 0.001) below that in control PCs on Days 1, 3, and 5, respectively. At similar intervals, a significant increase in recovery from hypotonic shock also was observed in AAA-incubated PCs. Further, Day 5 AAA-PCs demonstrated significantly higher morphology scores and O2 consumption than did control preparations. CONCLUSION: Buffy coat platelets prepared in suspended bags and stored in the presence of AAA demonstrate significantly reduced activation and enhanced functional and metabolic activity.     

Influence of a 12-hour, 22 degrees C holding period for buffy coats on the preparation of platelet concentrates stored in plasma

BACKGROUND: The preparation of platelet concentrates (PCs) from buffy coats (BCs) stored at room temperature is controversial, because of the strong metabolic activity of cells in BCs and the possible detrimental effect of neutrophil enzymes on platelets when the holding time before separation is prolonged. Despite good in vitro and in vivo behavior of BC-PCs stored in synthetic solution, little is known of the quality of BC-PCs stored in plasma. STUDY DESIGN AND METHODS: Comparison was made of PCs prepared from BCs held at 22 degrees C for 3 hours (3-hour BC- PCs) or overnight (12-hour BC-PCs) and stored in plasma. Platelet and white cell counts, pH, response to osmotic shock, and morphologic scores were determined on 20 PCs of each type. The decrease in dense granule and alpha granule content, a marker of platelet activation, were estimated by mepacrine counting and beta-thromboglobulin measurement, respectively (n = 8–10). Platelet function was studied in terms of aggregation and thromboxane production in response to various concentrations of collagen and thrombin (n = 8–17). PCs prepared from unstored BCs (n = 15) and from BCs held for 90 minutes (n = 15) were used as controls. RESULTS: Platelet yield was increased from 53 +/? 10 percent of donated platelets to 73 +/? 4 percent by increasing the BC holding time from 0 to 90 minutes to 3 hours (p < 0.001). Similar yields (7.8 +/? 1.8 vs. 7.9 +/? 2 × 10(10) platelets) and white cell contamination (0.9 +/? 0.8 vs. 1.0 +/? 0.9 × 10(7)) were obtained with 3-hour and 12-hour BC-PCs. At the end of the storage period (Day 5), all variables known to correlate with platelet survival in vivo were well maintained in both 3-hour and 12-hour BC-PCs: pH > or = 6.9, response to osmotic shock > or = 70 percent, and morphology scores always > or = 240. During storage, the dense granule content decreased moderately (30% after 5 days), whatever the conditions. By contrast, the total platelet beta-thromboglobulin content was better preserved in 12-hour BC-PCs than in 3-hour BC-PCs (p < 0.04). No significant differences were observed in collagen-induced aggregation and thromboxane production in the two PC preparations. However, aggregation responses to thrombin were higher in 12-hour BC-PCs on Day 5 of storage (p < 0.01). CONCLUSION: BCs can be held at 22 degrees C for up to 12 hours, with no detrimental effect on the quality of PCs stored for up to 5 days in plasma. Such a holding time might help overcome logistic problems in blood banks     

American Red Cross experience with routine testing for hepatitis B core antibody

The implementation of routine testing of blood donations for hepatitis B core antibody (anti-HBc) has allowed the characterization of the performance of the test in a large number of samples from apparently healthy individuals. This study reports the experience of the American Red Cross in testing 2.3 million donors for anti-HBc. The test protocol reproducibly identified a distinct population of donors. The anti-HBc-positive rate varied by region of the continental United States and by the time of year. In a case-control study, 85 percent of subsequent donations from anti-HBc-positive donors were anti-HBc positive. The predictions made in an earlier pilot study regarding the performance and impact of the test were borne out.     

Influence of the HLA-DRB1 locus on susceptibility and severity in rheumatoid arthritis

We examined HLA-DR genotype risk in 288 patients with rheumatoid arthritis who were carefully categorized for disease severity. Five hundred ethnically-matched bone-marrow donors were controls. A hierarchy of positive allelic associations was noted with DRB1*0401 (p < 10(-38), *0404,8 (p < 10(-43), *0405 (p < 10(-8), *10 (p < 10(-3) and *0101,2 (p < 10(-2), while DRB1*0403 was negatively associated (p = 0.02). The DRB1 genotype relative risks (and 95% CIs) for RA were: *0404,5,8/*0404,5,8 = 36.2 (15-87), *0401/*0404,5,8 = 31.3 (18-55), *401/*0401 = 18.8 (11-35), *0101,2/*0404,5,8 = 6.0 (2-14), *0101,2/*0401 = 6.4 (3-12), *0101,2/*0101,2 = 1.3 (0.3-6), *10/*0404,5,8 = 27.8 (5-148), *10/*0401 = 20.8 (5-89), *10/*0101,2 = 22.3 (5-96), *0404,5,8/DRX = 5.0 (3-8), *0401/DRX = 4.7 (3-7), *0101,2/DRX = 2.3 (1.4-4), *10/DRX = 3.4 (0.8-14). No significant correlation of DRB1 genotypes was found with severity of RA as judged by nodules or articular erosions.      

Characterization of cereal toxicity for celiac disease patients based on protein homology in grains

BACKGROUND AND AIMS: Celiac disease is caused by T-cell responses to wheat gluten-derived peptides. The presence of such peptides in other widely consumed grains, however, has hardly been studied. METHODS: We have performed homology searches to identify regions with sequence similarity to T-cell stimulatory gluten peptides in the available gluten sequences: the hordeins of barley, secalins of rye, and avenins of oats. The identified peptides were tested for T-cell stimulatory properties. RESULTS: With 1 exception, no identical matches with T-cell stimulatory gluten peptides were found in the other grains. However, less stringent searches identified 11 homologous sequences in hordeins, secalins, and avenins located in regions similar to those in the original gluten proteins. Seven of these 11 peptides were recognized by gluten-specific T-cell lines and/or clones from patients with celiac disease. Comparison of T-cell stimulatory sequences with homologous but non-T-cell stimulatory sequences indicated key amino acids that on substitution either completely or partially abrogated the T-cell stimulatory activity of the gluten peptides. Finally, we show that single nucleotide substitutions in gluten genes will suffice to induce these effects. CONCLUSIONS: These results show that the disease-inducing properties of barley and rye can in part be explained by T-cell cross-reactivity against gluten-, secalin-, and hordein-derived peptides. Moreover, the results provide a first step toward a rational strategy for gluten detoxification via targeted mutagenesis at the genetic level.     

Long-term frequent plasma exchange donation of cryoprecipitate

Plasma exchange donation accomplishes the selective donation of cryoprecipitate. It facilitates the repeated donation of large quantities of factor VIII by individual donors and reduces donor exposure for recipients. A highly motivated donor is described who has undergone 103 donations between May 1983 and March 1987, producing 359,460 IU of factor VIII and supplying all the factor VIII needed since August 1983 by his severely affected hemophiliac son, now age 14. The donor has remained in good health, and no significant abnormalities have been noted in hematologic, biochemical, immunologic, coagulation, and serum protein testing. Extensive experience with this donor suggests that repeated plasma-exchange donation is safe and can sometimes allow single-donor support of severe hemophiliacs.     

干细胞诱导分化为胰腺β细胞的研究现状

学术背景：干细胞体外诱导分化成的胰岛细胞可以发挥对血糖的生理性调节作用。大量研究证明可以从胚胎干细胞、胰腺干细胞、骨髓间充质干细胞、神经干细胞、肝脏干细胞或脐血干细胞等诱导分化出胰腺β细胞。目的：深入认识干细胞诱导分化为胰腺β细胞的研究现状。检索策略：由该论文的研究人员应用计算机检索Pubmed数据库1990—01，2006—07的相关文献，检索词“stem cell，differenliation，culture，pancreas”，并限定文章语言种类为English。同时计算机检索维普数据库2000-01，2006—07的相关文献，检索词“干细胞，胰岛，诱导分化”，并限定文章语言种类为中文。共检索到142篇文献，对资料进行初审，纳入标准：①与干细胞和干细胞向胰腺β细胞诱导分化相关的文章。②若内容相似，选取首次实验报告及近5年较权威杂志发表的文章。排除标准：重复或类似的研究。文献评价：文献的来源主要是通过对干细胞向胰腺β细胞诱导分化方面内容进行汇总分析。所选用的30篇文献中，1篇为综述，其余均为临床或基础实验研究。资料综合：胰岛移植是目前治疗Ⅰ型糖尿病和部分Ⅱ型糖尿病效果最理想的方法。由干细胞诱导分化得到的胰腺β细胞可以发挥调节血糖的作用，在许多小鼠糖尿病模型研究中起到了降低血糖的作用。目前用于诱导分化为胰腺β细胞的干细胞来源包括胚胎干细胞、胰腺干细胞、骨髓间充质干细胞、神经干细胞、肝脏干细胞或脐血干细胞等。在不同的干细胞诱导分化为胰腺β细胞的研究中，分为体内和体外两种诱导分化方法，而体外诱导法多数采用分步诱导的方式，也有部分实验利用基因技术的方法进行诱导。结论：不同来源的干细胞可以在体外通过多种方法诱导分化得到胰腺β细胞，但得到的胰腺β细胞数量及其诱导分化方法还有待进一步研究。