52 research outputs found

    Effect of Irradiation and/or Leucocyte Filtration on RBC Storage Lesions

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    Red blood cell (RBC) storage lesions have been shown to be associated with some adverse reactions; numerous studies have focused on the lesions caused by storage, and few data on the RBC storage lesions caused by prestorage treatments of leucocyte filtration and irradiation. In this study, we examined the changes related with the RBC storage lesions, including 2,3-diphosphatidylglyceric acid (2,3-DPG), pH, free hemoglobin (Hb), supernatant free K+ and Na+ concentration, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH). Along with the increasing storage time, decreases in 2, 3-DPG levels, pH and Na+ concentration, increases in K+ and free Hb concentrations, and significant morphological changes in RBC in all groups were found. The changes in the groups of irradiation, leucocyte filtration and the combined irradiation and leucocyte filtration were more significant than those in the untreated group. Meanwhile, the MCV levels of the three treated groups were significantly lower than those in the untreated group, while the MCH variations were significantly higher. Our results suggest that irradiation and leucocyte filtration before storage may aggravate blood storage lesions

    Transfer of MicroRNAs by Embryonic Stem Cell Microvesicles

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    Microvesicles are plasma membrane-derived vesicles released into the extracellular environment by a variety of cell types. Originally characterized from platelets, microvesicles are a normal constituent of human plasma, where they play an important role in maintaining hematostasis. Microvesicles have been shown to transfer proteins and RNA from cell to cell and they are also believed to play a role in intercellular communication. We characterized the RNA and protein content of embryonic stem cell microvesicles and show that they can be engineered to carry exogenously expressed mRNA and protein such as green fluorescent protein (GFP). We demonstrate that these engineered microvesicles dock and fuse with other embryonic stem cells, transferring their GFP. Additionally, we show that embryonic stem cells microvesicles contain abundant microRNA and that they can transfer a subset of microRNAs to mouse embryonic fibroblasts in vitro. Since microRNAs are short (21–24 nt), naturally occurring RNAs that regulate protein translation, our findings open up the intriguing possibility that stem cells can alter the expression of genes in neighboring cells by transferring microRNAs contained in microvesicles. Embryonic stem cell microvesicles may be useful therapeutic tools for transferring mRNA, microRNAs, protein, and siRNA to cells and may be important mediators of signaling within stem cell niches

    Sequestration and Tissue Accumulation of Human Malaria Parasites: Can We Learn Anything from Rodent Models of Malaria?

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    The sequestration of Plasmodium falciparum–infected red blood cells (irbcs) in the microvasculature of organs is associated with severe disease; correspondingly, the molecular basis of irbc adherence is an active area of study. In contrast to P. falciparum, much less is known about sequestration in other Plasmodium parasites, including those species that are used as models to study severe malaria. Here, we review the cytoadherence properties of irbcs of the rodent parasite Plasmodium berghei ANKA, where schizonts demonstrate a clear sequestration phenotype. Real-time in vivo imaging of transgenic P. berghei parasites in rodents has revealed a CD36-dependent sequestration in lungs and adipose tissue. In the absence of direct orthologs of the P. falciparum proteins that mediate binding to human CD36, the P. berghei proteins and/or mechanisms of rodent CD36 binding are as yet unknown. In addition to CD36-dependent schizont sequestration, irbcs accumulate during severe disease in different tissues, including the brain. The role of sequestration is discussed in the context of disease as are the general (dis)similarities of P. berghei and P. falciparum sequestration

    Specific Receptor Usage in Plasmodium falciparum Cytoadherence Is Associated with Disease Outcome

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    Our understanding of the basis of severe disease in malaria is incomplete. It is clear that pathology is in part related to the pro-inflammatory nature of the host response but a number of other factors are also thought to be involved, including the interaction between infected erythrocytes and endothelium. This is a complex system involving several host receptors and a major parasite-derived variant antigen (PfEMP1) expressed on the surface of the infected erythrocyte membrane. Previous studies have suggested a role for ICAM-1 in the pathology of cerebral malaria, although these have been inconclusive. In this study we have examined the cytoadherence patterns of 101 patient isolates from varying clinical syndromes to CD36 and ICAM-1, and have used variant ICAM-1 proteins to further characterise this adhesive phenotype. Our results show that increased binding to CD36 is associated with uncomplicated malaria while ICAM-1 adhesion is raised in parasites from cerebral malaria cases

    Tibor J. Greenwalt; in memoriam

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    Vesicles generated during storage of red cells are rich in the lipid raft marker stomatin.

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    Contains fulltext : 69440.pdf (publisher's version ) (Closed access)BACKGROUND: The release of vesicles by red blood cells (RBCs) occurs in vivo and in vitro under various conditions. Vesiculation also takes place during RBC storage and results in the accumulation of vesicles in RBC units. The membrane protein composition of the storage-associated vesicles has not been studied in detail. The characterization of the vesicular membrane might hint at the underlying mechanism of the storage-associated changes in general and the vesiculation process in particular. STUDY DESIGN AND METHODS: Vesicles from RBCs that had been stored for various periods were isolated and RBCs of the same RBC units were used to generate calcium-induced microvesicles. These two vesicle types were compared with respect to their size with atomic force microscopy, their raft protein content with detergent-resistant membrane (DRM) analysis, and their thrombogenic potential and activity with annexin V binding and thrombin generation, respectively. RESULTS: The storage-associated vesicles and the calcium-induced microvesicles are similar in size, in thrombogenic activity, and in membrane protein composition. The major differences were the relative concentrations of the major integral DRM proteins. In storage-associated vesicles, stomatin is twofold enriched and flotillin-2 is threefold depleted. CONCLUSION: These data indicate that a stomatin-specific, raft-based process is involved in storage-associated vesiculation. A model of the vesiculation process in RBCs is proposed considering the raft-stabilizing properties of stomatin, the low storage temperature favoring raft aggregation, and the previously reported storage-associated changes in the cytoskeletal organization

    Red cell concentrates of hemochromatosis patients comply with the storage guidelines for transfusion purposes.

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    Contains fulltext : 71238.pdf (publisher's version ) (Closed access)BACKGROUND: Therapeutic phlebotomy is the preferred treatment for iron overload associated with hemochromatosis. In the Netherlands, red blood cell concentrates (RCCs) from hemochromatosis patients are not used for transfusion purposes. In this study, their storage performance was compared with that of control donors as a first step in the evaluation of their potential usefulness for transfusion. STUDY DESIGN AND METHODS: RCCs were obtained from hemochromatosis patients and regular donors, either by apheresis or by whole-blood collection, and stored up to 50 days under routine Dutch blood bank conditions. Weekly samples were taken for determination of hematologic, biophysical, and biochemical variables. RESULTS: Most variables displayed the same storage-related changes in RCCs originating from hemochromatosis patients as in those from regular donors. In all RCCs, hemolysis remained well below the guideline limit of 0.8 percent for up to 6 weeks of storage, and the glucose concentration remained above the required 5 mmol per L up to 5 weeks of storage. After 4 weeks of storage, the mean ATP level remained above the required limit of 75 percent of the starting value in all RCCs as well. The major difference was a larger mean cell volume in hereditary hemochromatosis RBCs up to 50 days of storage. CONCLUSIONS: RCCs from hemochromatosis patients comply with the in vitro quality requirements for transfusion. This paves the way for the final step, namely, the establishment of the 24-hour RBC posttransfusion recovery
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