bluuebunny/crossref_metadata_2025_split huggingface dataset:
Vesicles generated during storage of red cells are rich in the lipid raft marker stomatin
Authors: Salzer Ulrich , Zhu Rong , Luten Marleen , Isobe Hirotaka , Pastushenko Vassili , Perkmann Thomas , Hinterdorfer Peter , Bosman Giel J.C.G.M. 
Published: February 2008

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.
Blood group genotyping for Jka/Jkb, Fya/Fyb, S/s, K/k, Kpa/Kpb, Jsa/Jsb, Coa/Cob, and Lua/Lub with microarray beads
Authors: Karpasitou Katerina , Drago Francesca , Crespiatico Loretta , Paccapelo Cinzia , Truglio Francesca , Frison Sara , Scalamogna Mario , Poli Francesca 
Published: February 2008

BACKGROUND: Traditionally, blood group typing has been performed with serologic techniques, the classical method being the hemagglutination test. Serotyping, however, may present important limitations such as scarce availability of rare antisera, typing of recently transfused patients, and those with a positive direct antiglobulin test. Consequently, serologic tests are being complemented with molecular methods. The aim of this study was to develop a low‐cost, high‐throughput method for large‐scale genotyping of red blood cells (RBCs).STUDY DESIGN AND METHODS: Single‐nucleotide polymorphisms associated with some clinically important blood group antigens, as well as with certain rare blood antigens, were evaluated: Jka/Jkb, Fya/Fyb, S/s, K/k, Kpa/Kpb, Jsa/Jsb, Coa/Cob, and Lua/Lub. Polymerase chain reaction (PCR)‐amplified targets were detected by direct hybridization to microspheres coupled to allele‐specific oligonucleotides. Cutoff values for each genotype were established with phenotyped and/or genotyped samples.RESULTS: The method was validated with a blind panel of 92 blood donor samples. The results were fully concordant with those provided by hemagglutination assays and/or sequence‐specific primer (SSP)–PCR. The method was subsequently evaluated with approximately 800 blood donor and patient samples.CONCLUSION: This study presents a flexible, quick, and economical method for complete genotyping of large donor cohorts for RBC alleles.
Evaluation of a new Trypanosoma cruzi antibody assay for blood donor screening
Authors: Gorlin Jed , Rossmann Susan , Robertson Gene , Stallone Fred , Hirschler Nora , Nguyen Kim‐Anh , Gilcher Ronald , Fernandes Helen , Alvey Stacey , Ajongwen Patience , Contestable Paul , Warren Harold 
Published: February 2008

BACKGROUND: This multicenter prospective study was designed to evaluate the performance characteristics of a new commercially available enzyme‐linked immunosorbent assay (ELISA) for the detection of antibodies to Trypanosoma cruzi in blood donors, the ORTHO T. cruzi ELISA Test System (Ortho‐Clinical Diagnostics).STUDY DESIGN AND METHODS: Assay specificity was evaluated among 40,665 serum and ethylenediaminetetraacetate (EDTA) plasma specimens from volunteer blood donors and 481 T. cruzi antibody–negative specimens from a high‐risk population. Sensitivity was evaluated among 106 T. cruzi‐infected subjects identified by parasite detection, among 93 radioimmunoprecipitation assay (RIPA)‐positive specimens from high‐risk subjects, and 662 specimens presumed positive for the presence of T. cruzi antibodies by serologic methods. Also assessed were the equivalence of serum and plasma as specimen sources, performance equivalence of automated and semiautomated processing methods, nonspecific reactivity in specimens from other disease states or clinical conditions, and assay precision.RESULTS: Assay specificity was 99.998 percent in volunteer blood donors and 99.4 percent among high‐risk subjects. Sensitivity was 100 percent among specimens positive by parasite detection, or by serologic methods, and 98.9 percent among RIPA‐positive specimens from high‐risk subjects. No differences were demonstrated between serum and plasma or between semiautomated and automated processing methods. Cross‐reactivity was observed with known positive leishmaniasis specimens. Total inter‐ and intraassay variability was less than 10 percent with both the automated and the semiautomated methods.CONCLUSION: The ORTHO T. cruzi ELISA Test System is an effective, qualitative assay for screening blood donors for immunoglobulin G antibodies to T. cruzi. The assay was licensed for donor screening by the FDA in December 2006.
Viability does not necessarily reflect the hematopoietic progenitor cell potency of a cord blood unit: results of an interlaboratory exercise
Authors: Brand Anneke , Eichler Hermann , Szczepiorkowski Zbigniew M. , Hess John R. , Kekomaki Riitta , McKenna David H. , Pamphilon Derwood , Reems JoAnna , Sacher Ronald A. , Takahashi Tsuneo A. , Van De Watering Leo M.G. ,   Biomedical Excellence for Safer Transfusion (BEST) Collaborative
Published: February 2008

BACKGROUND: Clinical transplant outcome with umbilical cord blood (UCB) as source of hematopoietic progenitor cells (HPCs) is, among other factors, determined by the total number of viable nucleated cells and/or CD34+ cells in the unit. Quantitative and qualitative losses by processing and cryopreservation and by thawing and washing before transfusion may occur, however. Another reason for a discrepancy between the number of cells in the unit released by the cord blood bank and found in the transplant center may be technical differences in cell counting methods between the two sites.STUDY DESIGN AND METHODS: With the collaborative group for Biomedical Excellence for Safer Transfusion (BEST), an interlaboratory exercise was conducted among nine sites for thawed UCB variables: total nucleated cells, CD34+ cells, viability, and HPC cultures. Three frozen UCB samples were shipped, with instructions for thawing, counting, and HPC plating.RESULTS: Unexpectedly samples arrived at all nine receiving centers without detectable hematopoietic progenitor colony‐forming cells. Nevertheless, wide interlaboratory ranges for viability were obtained. The proportion of viable cells was found higher with manual methods, but all viability assays used in the study overestimated functional progenitor cells.CONCLUSIONS: The results underscore the complexity of evaluation of frozen‐thawed cord blood cells and the need for standardization of assessment.
Persistence of allografts in the peritoneal cavity after prenatal transplantation in mice
Authors: Chen Jeng‐Chang , Chang Ming‐Ling , Muench Marcus O. 
Published: February 2008

BACKGROUND: In utero transplantation (IUT) is usually performed by intraperitoneal injection, but there is little information regarding the fate of intraperitoneally transplanted cells in the first weeks after transplantation.STUDY DESIGN AND METHODS: Fetal transplantation was performed in a B6D2F1 (C57BL/6 × DBA/2, H‐2b/d) into C57BL/6 (H‐2b) murine strain combination at the gestational age of 13 days by intraperitoneal injection of light‐density marrow cells. Adult C57BL/6 mice were used as the postnatal transplantation group. Donor cell levels in the peritoneum, peripheral blood, and various hematopoietic tissues were examined by flow cytometry.RESULTS: After fetal transplantation, peritoneal chimerism could be detected for 4 weeks, ranging from 0.02 to 23.2 percent with the highest median chimerism at 2 weeks after transplantation. Donor cells in the peripheral blood, spleen, marrow, liver, and thymus were usually low (<0.2%) with a tendency for higher donor cell levels at 2 weeks after transplantation. Engraftment in fetal peritoneum was primarily by myeloid and B cells with few T cells. In contrast, adult mice transplanted with haplogeneic marrow cleared nearly all donor cells from the peritoneum within a week of transplant. There was no evidence showing that donor cells had ever migrated to any hematopoietic tissues after adult transplantation.CONCLUSION: Despite limited engraftment in hematopoietic tissues, donor cells persisted in the peritoneum for 2 weeks or longer after fetal transplantation. The fetal peritoneum may serve as a sanctuary for foreign cells in the fetus, which may aid the development of novel cellular therapies for birth defects.
Estimated risk of Chikungunya viremic blood donation during an epidemic on Reunion Island in the Indian Ocean, 2005 to 2007
Authors: Brouard Cécile , Bernillon Pascale , Quatresous Isabelle , Pillonel Josiane , Assal Azzedine , De Valk Henriette , Desenclos Jean‐Claude 
Published: February 2008

BACKGROUND: Between 2005 and 2007, Chikungunya virus (CHIKV) caused a massive epidemic on Reunion Island with a major peak in the number of cases in February 2006. Blood donation was interrupted on the island in January 2006.STUDY DESIGN AND METHODS: Estimates of the mean risk of viremic blood donation on Reunion Island were computed for different phases of the epidemic. Calculations used CHIKV incidence estimates derived from sentinel surveillance, duration of viremia, and frequency of asymptomatic infection. Data on these two last parameters were initially based on hypotheses and subsequently obtained from studies carried out during the outbreak. The estimated risk was compared to the results of CHIKV nucleic acid testing (NAT) implemented for platelet (PLT) donations screening.RESULTS: Over the course of the outbreak, the mean risk was estimated at 132 per 100,000 donations. The risk peaked at 1,500 per 100,000 donations at the height of the outbreak in February 2006. In total, 47 blood donations would have been potentially viremic if blood collection had not been interrupted. During this period, an estimated 312,500 of 757,000 inhabitants had been infected by mosquito‐borne transmission. From January to May 2006, the estimated mean risk (0.7%) and observed risk on PLT donations (0.4%) were of the same order of magnitude.CONCLUSION: During this large outbreak, the estimated risk of viremic blood donation was high, but low compared to the risk of mosquito‐borne CHIKV transmission. The estimated risk was corroborated by the concordant results with the observed risk.
In vitro proliferation and differentiation of human CD34+ cells from peripheral blood into mature red blood cells with two different cell culture systems
Authors: Dorn Isabel , Lazar‐Karsten Pamela , Boie Stefanie , Ribbat Julika , Hartwig Dirk , Driller Birgit , Kirchner Holger , Schlenke Peter 
Published: February 2008

BACKGROUND:An in vitro erythropoiesis assay is a powerful tool for investigating red blood cell (RBC) development and diseases of the erythroid lineage. Most assays, however, failed in either proliferation or terminal differentiation. Here two liquid cultures (LCs) for in vitro generation of RBCs from peripheral blood CD34+ cells were compared.STUDY DESIGN AND METHODS:Granulocyte–colony‐stimulating factor–mobilized CD34+ cells were cultured for 16 days in a two‐phase LC (2P‐LC; Days 1‐8, stem cell factor [SCF], erythropoietin [EPO], insulinlike growth factor [IGF]‐1, and steroids; Days 9‐16, EPO and insulin) and for 21 days in a three‐phase LC (3P‐LC; Days 1‐7, SCF, thrombopoetin, and Flt3‐ligand; Days 8‐14, SCF, EPO, and IGF‐1; Days 15‐21, EPO and IGF‐1). Maturation was analyzed by flow cytometry (CD36, CD71, glycophorin A [GPA]) and microscopy.RESULTS:In the 2P‐LC, cell numbers increased from 0.5 × 106to 25.7 × 106 ± 15.1 × 106cells per mL. More than 95 percent were GPA+ and showed morphologic characteristics of normoblasts (52 ± 15%) and enucleated reticulocytes (43 ± 18%). In the 3P‐LC, a higher overall proliferation to 55.7 × 106 ± 37.7 × 106cells per mL was achieved (p < 0.05). This was also accompanied by a high degree of normoblasts (36 ± 16%) and reticulocytes (48 ± 24%). The amount of GPA+ cells was slightly lower (88.4 ± 16.4%), associated with a significantly higher contamination by nonerythroid cells (15.8 ± 19.3% vs. 3.9 ± 2.9%, p < 0.05).CONCLUSION:Both LCs were able to generate fully matured RBCs and represent powerful tools for fundamental research in erythroid development and diseases targeting the erythroid lineage. A slightly higher proliferation was achieved in the 3P‐LC. This was associated with a limited homogeneity due to more nonerythroid cells, however. Therefore the 2P‐LC is favored, also saving additional culture days and growth factors.
Release of immune modulation factors from platelet concentrates during storage after photochemical pathogen inactivation treatment
Authors: Cognasse Fabrice , Osselaer Jean‐Claude , Payrat Jean Marc , Chavarin Patricia , Corash Laurence , Garraud Olivier 
Published: February 2008

BACKGROUND: Blood platelets (PLTs) are critical for hemostasis, and they contain biologically active constituents with the potential to modulate inflammatory responses. This study examined the effects of photochemical pathogen inactivation treatment (PCT) on the release of cytokines and/or chemokines from PLT components.STUDY DESIGN AND METHODS: Double‐dose apheresis PLT components were suspended in plasma–PLT additive solution mixtures and divided into paired therapeutic units. One unit served as an untreated control and the other unit was treated with PCT. PLT concentrations, pH, and levels of cytokines and/or chemokines (CD62p, platelet‐derived growth factor‐AB, interleukin [IL]‐8, soluble CD40 ligand [sCD40L], IL‐1β, and tumor necrosis factor α) were measured during 7 days of storage in PLT component supernatants and PLT lysates.RESULTS: PLT content, pH, and cytokine and/or chemokine content and release from PLT component prepared with PCT were not different (p > 0.05) from paired control components during storage. Levels of sCD40L, however, increased significantly during storage while decreasing in parallel within PLT lysates, although no differences were detected between paired PCT and control PLT component.CONCLUSION: PCT did not increase the release or secretion of PLT chemokines and/or cytokines over a 7‐day period compared to conventional PLT component.
Use of molecular markers in participatory plant breeding: assessing the genetic variability in cotton populations bred by farmers
Authors: Lançon J. , Pichaut J.‐P. , Djaboutou M. , Lewicki‐Dhainaut S. , Viot C. , Lacape J.‐M. 
Published: February 2008

AbstractIn participatory plant breeding, farmers are involved in simple selection schemes that are not suitable for assessing genetic variability in the segregating populations. We propose to use information derived from molecular marker analyses to help monitoring such populations. In this study, we used three indicators to compare genetic variability in eight genetic structures, that is three plant populations selected by farmers over five generations, three nonselected populations and two commercial varieties. The three indicators were the polymorphic locus rate, heterozygosity rate and dissimilarity index. The results highlighted that the genetic variability decreased more with farmers’ selection than with environmental factors. The breeding process was not complete because genetic variability in the selected populations was midway between that of the nonselected populations and that of the commercial varieties monitored. The three proposed indicators were relevant for describing the studied populations. They could be interpreted according to a grid drawn up on the basis of the results of the present study.
Seed Dispersal and Ingestion of Insect‐Infested Seeds by Black Howler Monkeys in Flooded Forests of the Parana River, Argentina
Authors: Bravo Susana Patricia 
Published: February 2008

ABSTRACTAll howler monkey species (Alouatta spp.) have a folivorous–frugivorous diet. Howler monkeys are reported to be seed dispersers in several areas, including black howlers (Alouatta caraya), which are important seed dispersers in northern Argentinean forests. The goal of this work was to study the three‐way interaction between insects, seeds, and black howlers, and assess the functional significance of this tri‐trophic interaction for seed dispersal. I determined through direct observation that fruits of species with a high proportion of insect infestation were important components of howler monkey diet. Ocotea diospyrifolia seeds from fresh faeces of black howlers contained dead larvae, but seeds were still able to germinate. Seeds in which larvae had reached an advanced stage of development did not germinate. Larvae of infested Eugenia punicifolia fruits were killed by digestion when they occurred in the pulp early in the fruiting season, but were dispersed alive with seeds later in the season. Banara arguta fruits contained both healthy and infested seeds; infested seeds were destroyed during digestion, while healthy seeds were dispersed. Black howlers' ingestion of infested fruits could result in the: (1) killing of larvae and dispersion of healthy seeds; (2) spread of larvae; or (3) destruction of infested seeds. This will depend on the relationship between the time at which fruit is consumed by black howlers, the time at which insect infestation occurs, and also probably on the hardness of the seed coat and the seed–insect size ratio.
Green fluorescent protein incorporation by mouse myoblasts may yield false evidence of myogenic differentiation of human haematopoietic stem cells
Authors: Di Castro A. , Bonci D. , Musumeci M. , Grassi F. 
Published: February 2008

AbstractAims:  Haematopoietic CD34+ stem cells are able to differentiate into skeletal muscle, a potentially invaluable tool for treating degenerative diseases such as muscular dystrophy. However, some studies argue that the differentiative potential of these cells might have been overestimated. In vitro studies provide a controlled environment in which to investigate this point.Methods:  CD34+ stem cells from human peripheral blood, labelled with green fluorescent protein (GFP), were co‐cultured with mouse myogenic C2C12 cells. The functional properties of mononucleated GFP+ cells were determined using electrophysiological techniques and were related to protein profiling determined by immunofluorescence staining and single‐cell RT‐PCR. Mouse mesoangioblasts co‐cultured with human myotubes provided methodological controls.Results:  After 2–4 days, mononucleated adherent GFP+ cells showed acetylcholine‐evoked current responses, typical of myogenic cells, as if stem cells had integrated into the host environment. In contrast to this hypothesis, human nuclei could not be detected in adherent GFP+ cells by immunofluorescence. Moreover, single‐cell RT‐PCR showed that adherent GFP+ cells responsive to acetylcholine expressed mouse markers while loose unresponsive GFP+ cells were of human origin. The transcripts of the human α1 subunit of the acetylcholine muscle receptor were not amplified in co‐cultures.Conclusion:  Single‐cell analysis of functional properties combined with other markers revealed that, under the co‐culture conditions used, GFP was transferred from human CD34+ stem cells to C2C12 myoblasts by mechanisms unrelated to myogenic stem cell differentiation. Our results emphasize the need for careful controls using several markers when investigating the myogenic differentiation of circulating stem cells.
Statistical simulation of flood variables: incorporating short‐term sequencing
Authors: Cai Y. , Gouldby B. , Hawkes P. , Dunning P. 
Published: February 2008

AbstractThe pluvial and fluvial flooding in the United Kingdom over the summer of 2007 arose as a result of anomalous climatic conditions that persisted for over a month. Gaining an understanding of the sequencing of storm events and representing their characteristics within flood risk analysis is therefore of importance. This paper provides a general method for simulating univariate time series data, with a given marginal extreme value distribution and required autocorrelation structure, together with a demonstration of the method with synthetic data. The method is then extended to the multivariate case, where cross‐variable correlations are also represented. The multivariate method is shown to work well for a two‐variable simulation of wave heights and sea surges at Lerwick. This work was prompted by an engineering need for long time series data for use in continuous simulation studies where gradual deterioration is a contributory factor to flood risk and potential structural failure.
Molecular Characterization of New Natural Hybrids of
Saccharomyces cerevisiae
and
S
.
kudriavzevii
in Brewing
Authors: González Sara S. , Barrio Eladio , Querol Amparo 
Published: February 2008

ABSTRACT

We analyzed 24 beer strains from different origins by using PCR-restriction fragment length polymorphism analysis of different gene regions, and six new
Saccharomyces cerevisiae
×
Saccharomyces kudriavzevii
hybrid strains were found. This is the first time that the presence in brewing of this new type of hybrid has been demonstrated. From the comparative molecular analysis of these natural hybrids with respect to those described in wines, it can be concluded that these originated from at least two hybridization events and that some brewing hybrids share a common origin with wine hybrids. Finally, a reduction of the
S. kudriavzevii
fraction of the hybrid genomes was observed, but this reduction was found to vary among hybrids regardless of the source of isolation. The fact that 25% of the strains analyzed were discovered to be
S. cerevisiae
×
S. kudriavzevii
hybrids suggests that an important fraction of brewing strains classified as
S. cerevisiae
may correspond to hybrids, contributing to the complexity of
Saccharomyces
diversity in brewing environments. The present study raises new questions about the prevalence of these new hybrids in brewing as well as their contribution to the properties of the final product.