bluuebunny/crossref_metadata_2025 huggingface dataset:
Lithospheric boudinage in the Western Mediterranean back‐arc basin
Authors: Gueguen E. , Doglioni C. , Fernandez M.
Published: November 2003

The western Mediterranean is composed of irregular troughs formed as back‐arc basins in the hanging wall of the W‐directed Apenninic subduction which retreated eastward during Neogene and Quaternary times. The basins are progressively younger toward the east, ageing from late Oligocene–early Miocene (Valencia, Provençal, Alboran and Algerian basins), to middle Miocene–Pleistocene (Tyrrhenian Sea). The basins isolated boudins of continental lithosphere, the Sardinia–Corsica block representing the largest. The boudinage has a wavelength of 100–400 km and facilitated stretching of the continental lithosphere with formation of new oceanic crust in the Provençal, Algerian, Vavilov and Marsili basins. The boudins developed both in the earlier Alpine–Betic orogen (Alboran basin) and in its foreland (Provençal and Valencia troughs). The extension appears clearly asymmetric due to its eastward polarity, accommodated by E‐dipping master low‐angle normal faults. Moreover the thinning shows variations in boudinage wavelength and is characterized by several along‐strike transfer zones and heterogeneities. The western Mediterranean back‐arc setting is comparable with Atlantic and western Pacific back‐arc basins associated with W‐directed subduction zones that show similar large‐scale lithospheric boudinage.
Moons’ magnetism: from Io’s and Ganymede’s present magnetic signatures to the ancient lunar dynamo
Authors: Rochette P.
Published: November 2003

Intrinsic magnetic fields, corresponding to virtual axial dipole moments of the order of 1020 Am2, have recently been evidenced for the Jovian satellites Io and Ganymede. By reviewing the rock magnetic and palaeomagnetic properties together with the history of Io, the hypothesis of a moment due to induced or remanent magnetization of crustal rocks acquired within the ambient Jovian field is clearly eliminated. This demonstration is all the more valid for Ganymede, which experiences a much lower Jovian field. The demonstration of a present dynamo action for these Jovian moons, possibly sustained by Jupiter through tidal heating and background magnetic field, may be an actualistic model for the early lunar history. The hypothesis of a lunar dynamo, active at 3–4 Ga, seems to be strongly supported by this analogy.
The crustal structure of the western Po plain: reconstruction from integrated geological and seismic data
Authors: Biella G. , Polino R. , Franco R. de , Rossi P.M. , Clari P. , Corsi A. , Gelati R.
Published: November 2003

A reflection/refraction seismic experiment performed in 1991 in the western Po plain gave basic data to constrain the interpretation of the crustal structures across the Alps/Apennines junction zones. Two different seismic domains, north and south of the western supposed prosecution of the Villalvernia‐Varzi line, are evidenced from the interpretation of the data. The boundary between the two domains is characterized by strong lateral variations from southern high to northern low velocity layers. The northward abrupt deepening of the refractor/reflector basement is followed at depth by a similar deepening of the crust/mantle boundary. The geological interpretation evidences domains with coherent and independent evolution at surface level juxtaposed along oblique discontinuities cutting across the crust. A peculiar feature is the presence of both crust and mantle north‐verging wedges into the crustal structure and the overthrust at depth of the ‘alpine’ metamorphic crust onto the ‘apenninic’ nappes (Monferrato region).
Seismic mapping of crustal structures beneath the Indus‐Yarlung Suture, Tibet
Authors: Zhao W. , Mechie J. , Guo J. , Wenzel F. , Meissner R. , Ratschbacher L. , Steentoft H. , Husen S. , Brauner H.‐J. , Jiang D. , Frisch W. , Hauff S.‐F.
Published: November 2003

Records of densely spaced shots along the Sino‐US reflection line INDEPTH II at offsets between 70 and 130 km parallel to the main profile provide an image of the crust straddling the Indus‐Yarlung suture. The major features are prominent reflections at about 20 km depth beneath and extending out to about 20–30 km north and south of the surface exposure of the suture, and north‐dipping reflectors north of the suture. Various interpretations for the reflections are possible. (i) They represent a decollement, possibly of the Gangdise thrust system. In this scenario, the surface expression of the Gangdise thrust as mapped in eastern south Tibet is a splay with the decollement continuing southwards and either ending as a blind thrust or ramping up as one of the thrusts within the northernmost Tethyan shelf sequence. (ii) The reflections represent fabrics within gneisses, partly obliterated by intrusions reaching various levels of the crust. The reflection bands may be interpreted in terms of deformation or sedimentary structures belonging to the Indian crust, the accretionary complex, and the basement of the Gangdise belt. The intrusions could be related to the Tethyan leucogranites south of the suture (Rinbung leucogranite), and to the Gangdise magmatic arc to the north of the suture. (iii) The reflections represent a fortuitous coincidence of different features north and south of the suture. South of the suture, the reflections may record the basement–cover interface of the Indian crust or a thrust system in the Tethyan shelf. North of the suture, they may comprise different levels within the Gangdise belt and its basement. Although it is not possible to discriminate between the suggested scenarios without additional information, the seismic mapping points to the importance of post‐collisional (Oligocene–Miocene) tectonics, which reshaped the suture.
Two stages of zircon and monazite growth in anatectic leucogneiss: SHRIMP constraints on the duration and intensity of Pan‐African metamorphism in Prydz Bay, East Antarctica
Authors: Fitzsimons I.C.W. , Kinny P.D. , Harley S.L.
Published: November 2003

SHRIMP U–Pb data from two samples of foliated anatectic leucogneiss bracket the high‐grade metamorphic history of metasediments from southern Prydz Bay, East Antarctica. Magmatic zircons in these two samples have mean crystallization ages of 535 ± 13 and 536 ± 35 Myr, and provide a good estimate for the timing of peak metamorphism and partial melting. Low‐Th monazite grains in the same samples have mean ages of 528 ± 4 and 527 ± 11 Myr, which are within error of the zircon ages but may reflect a lower blocking temperature. High‐Th monazite rims with younger mean ages of 518 ± 3 and 512 ± 13 Myr occur around low‐Th cores in both samples, and were precipitated from hydrous fluids released during the crystallization of residual melts. These magmatic fluids were channelled along the retrograde foliation present in both samples. All mineral assemblages and fabrics preserved in the metasediments reflect intense Pan‐African metamorphism and deformation, associated with high degrees of partial melting and significant vertical displacements of the crust. Continental reconstructions assuming limited Pan‐African tectonism in East Antarctica should be reassessed in the light of these new data.
Variscan granites of the Pyrenees revisited: their role as syntectonic markers of the orogen
Authors: Gleizes G. , Leblanc D. , Bouchez J.L.
Published: November 2003

Structural studies of Variscan plutons of the Pyrenees demonstrate their syn‐orogenic character, contrasting with their post‐orogenic Permian Rb‐Sr datation. The Bassiès and Mont‐Louis‐Andorra plutons were emplaced before the main Variscan D2 phase and behaved as rigid markers during D2. Emplacement of the Néouvielle pluton was coeval with D2 as attested by its behaviour, first as a deformable body, then as a rigid body. The structures in plutons, combined with those of the country rocks, demonstrate that the whole Pyrenees acted as a large dextral ductile shear zone during the D2 main phase. This conclusion points out to the interest of the granitoids as kinematic markers.
Arabidopsis AtGSTF2 is regulated by ethylene and auxin, and encodes a glutathione S‐transferase that interacts with flavonoids
Authors: Smith Aaron P. , Nourizadeh Saeid D. , Peer Wendy A. , Xu Jinghuai , Bandyopadhyay Anindita , Murphy Angus S. , Goldsbrough Peter B.
Published: November 2003

Expression of the Arabidopsis glutathione S‐transferase (GST) gene AtGSTF2 is induced by several stimuli, but the function of this GST remains unknown. We demonstrate that AtGSTF2 expression is also induced by glutathione, paraquat, copper, and naphthalene acetic acid (NAA) via a mechanism independent of ethylene perception, as determined by analysis of the ethylene‐insensitive etr1 mutant. Deletion analyses identified two promoter regions important for regulation of AtGSTF2 expression in response to several of these inducers. Previous studies have suggested that AtGSTF2 interacts with indole‐3‐acetic acid (IAA) and the auxin transport inhibitor 1‐N‐naphthylphthalamic acid (NPA). We show that recombinant AtGSTF2 directly binds IAA, NPA, and the artificial auxin NAA. As NPA may act as an endogenous flavonoid regulator of auxin transport, competition between NPA and flavonoids for binding to AtGSTF2 was examined. Both quercetin and kaempferol competed with NPA for AtGSTF2 binding, indicating that all three compounds bind AtGSTF2 at the same site. In transgenic Arabidopsis seedlings, AtGSTF2::GUS expression occurred at the root–shoot transition zone and was induced in this region, as well as at the root distal elongation zone, after treatment with IAA. In wild‐type seedlings, AtGSTF2 is localized near the plasma membrane of cells in the root–shoot transition zone. However, both AtGSTF2::GUS expression and localization of AtGSTF2 protein were disrupted in flavonoid‐deficient tt4 seedlings. Our results indicate that AtGSTF2 is involved not only in stress responses but also in development under normal growth conditions.
Regiospecific hydroxylation of isoflavones by cytochrome P450 81E enzymes from Medicago truncatula
Authors: Liu Chang‐Jun , Huhman David , Sumner Lloyd W. , Dixon Richard A.
Published: November 2003

Mining of Medicago truncatula EST databases and screening of a root cDNA library led to the identification of three cytochrome P450 81E subfamily members. Two were functionally characterized by expression in yeast. The recombinant enzymes in yeast microsomes utilized the same isoflavone substrates, but produced different products hydroxylated at the 2′ and/or 3′ positions of the B‐ring. When transiently expressed in alfalfa leaves, green fluorescent protein (GFP) fusions of the isoflavone 2′‐ and 3′‐hydroxylases localized to the endoplasmic reticulum. The isoflavone 2′‐hydroxylase was functional when expressed in Arabidopsis. Differential tissue‐specific and biotic/abiotic stress‐dependent expression patterns were observed for the isoflavone 2′‐hydroxylase and 3′‐hydroxylase genes, suggesting differential involvement of 2′‐ and 3′‐hydroxylated isoflavonoids in pathogen defense and insect‐induced responses, respectively, in Medicago.
Virulence systems of Pseudomonas syringae pv. tomato promote bacterial speck disease in tomato by targeting the jasmonate signaling pathway
Authors: Zhao Youfu , Thilmony Roger , Bender Carol L. , Schaller Andreas , He Sheng Yang , Howe Gregg A.
Published: November 2003

Pseudomonas syringae pv. tomato strain DC3000 (Pst DC3000) causes bacterial speck disease on tomato. The pathogenicity of Pst DC3000 depends on both the type III secretion system that delivers virulence effector proteins into host cells and the phytotoxin coronatine (COR), which is thought to mimic the action of the plant hormone jasmonic acid (JA). We found that a JA‐insensitive mutant (jai1) of tomato was unresponsive to COR and highly resistant to Pst DC3000, whereas host genotypes that are defective in JA biosynthesis were as susceptible to Pst DC3000 as wild‐type (WT) plants. Treatment of WT plants with exogenous methyl‐JA (MeJA) complemented the virulence defect of a bacterial mutant deficient in COR production, but not a mutant defective in the type III secretion system. Analysis of host gene expression using cDNA microarrays revealed that COR works through Jai1 to induce the massive expression of JA and wound response genes that have been implicated in defense against herbivores. Concomitant with the induction of JA and wound response genes, the type III secretion system and COR repressed the expression of pathogenesis‐related (PR) genes in Pst DC3000‐infected WT plants. Resistance of jai1 plants to Pst DC3000 was correlated with a high level of PR gene expression and reduced expression of JA/wound response genes. These results indicate that COR promotes bacterial virulence by activating the host's JA signaling pathway, and further suggest that the type III secretion system might also modify host defense by targeting the JA signaling pathway in susceptible tomato plants.
OsHAP3 genes regulate chloroplast biogenesis in rice
Authors: Miyoshi Kazumaru , Ito Yukihiro , Serizawa Akiko , Kurata Nori
Published: November 2003

We have isolated three genes that potentially encode a HAP3/nuclear factor‐YB (NF‐YB)/CCAAT binding factor‐A (CBF‐A) subunit of a CCAAT‐box binding complex in rice (Oryza sativa), and named them OsHAP3A, OsHAP3B and OsHAP3C. These genes were expressed in various organs including leaves. In the transgenic rice plants with antisense or RNAi construct of OsHAP3A, reduced expression of not only OsHAP3A but also OsHAP3B and OsHAP3C was observed. These plants had pale green leaves, in which the amount of chlorophyll was reduced and chloroplasts were degenerated. Lamella was not well developed and accumulation of starch was not detected. The degenerated chloroplast formation was accompanied by reduced expression of nuclear‐encoded photosynthesis genes such as RBCS and CAB, while expression of chloroplast‐encoded genes was not affected or rather increased. These results suggest that one or more OsHAP3 genes regulate the expression of nuclear‐encoded chloroplast‐targeted genes and normal development of chloroplasts.
pl‐bol3, a complex allele of the anthocyanin regulatory pl1 locus that arose in a naturally occurring maize population
Authors: Pilu Roberto , Piazza Paolo , Petroni Katia , Ronchi Angela , Martin Cathie , Tonelli Chiara
Published: November 2003

The pl1 gene encodes a MYB‐related transcriptional activator committed to the regulation of anthocyanin biosynthesis in maize. Here, we report the genetic and molecular characterisation of pl‐bol3, an Andean allele displaying features that make it different from all the known pl1 alleles. pl‐bol3 has partial, light‐independent expression, and it is active mainly in the juvenile phase of growth. It has a complex molecular structure, containing multiple pl1 gene copies, thus being the first complex locus discovered in the c1/pl1 family. Although the composite genes of the complex locus encode proteins identical to other functional PL1 proteins, the putative promoters of the pl‐bol3 gene are different from the promoters of Pl‐Rhoades (Pl‐Rh) and pl1 sun‐red alleles. The intensity and the tissue specificity of anthocyanin production directed by pl‐bol3 differ significantly from that of Pl‐Rh and the original pl‐W22, and are specified by the interaction of pl‐bol3 with the different r1/b1 gene family members and the competence of pl‐bol3 to different pigment tissues. This allele represents a natural example of gene duplication and diversification of expression, giving rise to a significant change in phenotype and, in this way, is analogous to the complex r1 locus in maize. Analysis of the pl‐bol3 allele contributes to understanding the generation of diversity associated with multiple‐copy genes and the molecular basis of allele‐specific gene expression.
A nucleus‐encoded factor, CRR2, is essential for the expression of chloroplast ndhB in Arabidopsis
Authors: Hashimoto Mihoko , Endo Tsuyoshi , Peltier Gilles , Tasaka Masao , Shikanai Toshiharu
Published: November 2003

The chloroplast NDH complex, NAD(P)H dehydrogenase, reduces the plastoquinone pool non‐photochemically and is involved in cyclic electron flow around photosystem I (PSI). A transient increase in chlorophyll fluorescence after turning off actinic light is a result of NDH activity. We focused on this subtle change in chlorophyll fluorescence to isolate nuclear mutants affected in chloroplast NDH activity in Arabidopsis by using chlorophyll fluorescence imaging. crr2‐1 and crr2‐2 (chlororespiratory reduction) are recessive mutant alleles in which accumulation of the NDH complex is impaired. Except for the defect in NDH activity, photosynthetic electron transport was unaffected. CRR2 encodes a member of the plant combinatorial and modular protein (PCMP) family consisting of more than 200 genes in Arabidopsis. CRR2 functions in the intergenic processing of chloroplast RNA between rps7 and ndhB, which is possibly essential for ndhB translation. We have determined the function of a PCMP family member, indicating that the family is closely related to pentatrico‐peptide PPR proteins involved in the maturation steps of organellar RNA.
Cellulose microfibril alignment recovers from DCB‐induced disruption despite microtubule disorganization
Authors: Himmelspach Regina , Williamson Richard E. , Wasteneys Geoffrey O.
Published: November 2003

Cellulose microfibril deposition patterns define the direction of plant cell expansion. To better understand how microfibril alignment is controlled, we examined microfibril orientation during cortical microtubule disruption using the temperature‐sensitive mutant of Arabidopsis thaliana, mor1‐1. In a previous study, it was shown that at restrictive temperature for mor1‐1, cortical microtubules lose transverse orientation and cells lose growth anisotropy without any change in the parallel arrangement of cellulose microfibrils. In this study, we investigated whether a pre‐existing template of well‐ordered microfibrils or the presence of well‐organized cortical microtubules was essential for the cell to resume deposition of parallel microfibrils. We first transiently disrupted the parallel order of microfibrils in mor1‐1 using a brief treatment with the cellulose synthesis inhibitor 2,6‐dichlorobenzonitrile (DCB). We then analysed the alignment of recently deposited cellulose microfibrils (by field emission scanning electron microscopy) as cellulose synthesis recovered and microtubules remained disrupted at the mor1‐1 mutant's non‐permissive culture temperature. Despite the disordered cortical microtubules and an initially randomized wall texture, new cellulose microfibrils were deposited with parallel, transverse orientation. These results show that transverse cellulose microfibril deposition requires neither accurately transverse cortical microtubules nor a pre‐existing template of well‐ordered microfibrils. We also demonstrated that DCB treatments reduced the ability of cortical microtubules to form transverse arrays, supporting a role for cellulose microfibrils in influencing cortical microtubule organization.
Bacterial contamination of peripheral blood progenitor cells for transplantation
Authors: Jestice H. K. , Farrington M. , Hunt C. , Matthews I. , Scott M. A. , Foreman J. , Marcus R. E.
Published: November 2003

Peripheral blood progenitor cells (PBPC) were obtained from 128 apheresis harvests on 64 patients and were tested in duplicate for microbiological contamination (1) after collection and (2) after thawing, following processing and cryopreservation. In this study we have attempted to improve the monitoring of contamination in peripheral blood progenitor cell collections by identifying exogenous contamination that probably originated from the testing laboratory and is therefore not clinically significant. We found no contamination in 82% of harvests, 1.6% of harvests to be significantly contaminated and organisms were isolated from 16.4% that were assessed as clinically nonsignificant.  Our experience indicates that the choice of microbiological methods will influence the results and their clinical relevance. No samples were positive by direct culture. We recommend that sampling should be performed at more than one stage during the procedure and that initially only the post‐thaw samples be analysed. Testing should be performed by enrichment culture in duplicate only and if positive to aid interpretation the post‐collection sample should then be cultured.  No patient given nonsignificantly contaminated graft without antibiotic cover suffered infection from the identified organism. The incidence of significant contamination was low and we recommend that in these cases PBPC grafts can be infused safely provided prophylactic antibiotic cover is given.
Frequency and specificity of platelet‐specific alloantibodies in HLA‐immunized haematologic–oncologic patients
Authors: Schnaidt M. , Northoff H. , Wernet D.
Published: November 2003

The frequency and specificity of platelet‐alloantibodies to human platelet antigens (HPA) ‐1, ‐3 and ‐5 was investigated in 59 multitransfused, HLA‐immunized patients. Using the MAIPA test (monoclonal antibody specific immobilization of platelet antigens) platelet alloantibodies could be demonstrated in 10 (17%) patients.  In one patient the antibody was present prior to any transfusions and probably induced by multiple previous pregnancies. This antibody was directed to HPA‐5b. The remaining nine antibodies were found in patients (n = 36) with HLA‐antibodies reacting with over 95% of unselected lymphocytes. In these patients the target antigens were HPA‐1b in six, HPA‐3a in one and both antigens in two patients.  Our findings demonstrate platelet alloimmunization induced by transfusions to be restricted to patients with high HLA‐immunization. 25% of these patients (9/36) show platelet‐specific antibodies, primarily HPA‐1b.
Transcytosis of IgG anti‐D by human term trophoblast cells in culture
Authors: Kumpel B. M. , Sooranna S. R.
Published: November 2003

Placental trophoblast cells were cultured on filters that allow access to medium bathing the apical and basal surfaces of cells. Purified human IgG was added to either the apical or the basal chambers and sampled at intervals of up to 100 min for determination of IgG concentration by ELISA. Using this experimental system, transport of IgG1 and IgG3 monoclonal anti‐D, affinity‐purified polyclonal anti‐D and human polyclonal IgG were shown to occur primarily in the apical to basal (i.e. maternal to fetal) direction. The overall transport of monoclonal and polyclonal anti‐D was less than 10% in 45 min, several times lower than that of IgG. There was no difference in the rate or percentage of transport between IgG1 (BRAD‐5) and IgG3 (BRAD‐3) monoclonal anti‐D. The possibility that the Fc receptor mediating transcytosis of IgG anti‐D through human trophoblast cells in culture is the placental hFcRn is proposed.
From maximum surgical blood ordering schedule to unlimited computer crossmatching: evolution of blood transfusion for surgical patients at a tertiary hospital in Hong Kong
Authors: Chan A. H. C. , Chan J. C. M. , Wong L. Y. F. , Cheng G.
Published: November 2003

The maximum surgical blood ordering schedule (MSBOS) was developed to permit efficient use of blood stocks and reduce blood wastage due to outdating. Success of the MSBOS required full cooperation between surgeons, anaesthetists, blood bankers and house officers. However, clinical staff sometimes disregarded the system or distrusted the ability of the laboratory to provide blood in an emergency. This led to frustration and wastage of laboratory staff effort. We developed a computer crossmatch system that could overcome the shortcomings of the MSBOS. The advantages of this system are reported.
Antibodies to human major histocompatibility complex products inhibit Fcγ receptors types I and II
Authors: Neppert J. , Jungi T. W.
Published: November 2003

We have analysed the effect of polyclonal and monoclonal antibodies of distinct IgG isotypes directed against products of the human major histocompatibility complex (MHC) on the function of Fcγ receptors types I (FcγRI) and II (FcγRII). Human anti‐D sensitized red blood cells (RBC), selectively binding to FcγRI, and bovine or murine IgG1 (bIgG1 or mIgG1) sensitized RBC, selectively binding to FcγRII, were employed as targets for the effector cell in order to distinguish the inhibition of both types. Using these targets it could be shown that human antibodies to products of the human MHC or murine monoclonal antibodies with the same specificity and mIgG2a isotype inhibited both the FcγRI‐ and the FcγRII‐mediated function. In contrast, murine monoclonal antibodies with the same specificity but of the mIgG1 isotype only inhibited the FcγRII‐mediated function. In a control experiment, human and murine antibodies to products of the MHC did not impair the FcγR‐independent phagocytosis of Saccharomyces by monocytes and neutrophils.  The present study suggests that this mechanism involves at least two different Fcγ receptors.
Development of a PCR‐based diagnostic assay for the determination of KEL genotype in donor blood samples
Authors: Murphy M. T. , Fraser R. H. , Goddard J. P.
Published: November 2003

The polymorphism which determines expression of Kell and Cellano antigens on the red‐cell surface has been reported to be a single C→T nucleotide substitution at residue 701 where T codes for the presence of Kell antigen. This was confirmed by the direct automated sequencing of PCR products amplified from individuals of known Kell phenotype. The substitution creates a Bsm I restriction enzyme site and this has formed the basis for the development of a PCR‐based diagnostic assay for the determination of Kell phenotype in samples of donor blood. The assay is based on RFLP analysis of coamplified PCR products, one of which spans the K/k polymorphic site, and one control fragment which contains a Bsm I site. Digestion of the PCR products with Bsm I restriction enzyme and subsequent gel analysis of the digest allowed unequivocal determination of the K/k status of all of the samples tested.
The distribution of the MiIII (GP.Mur) phenotype among the population of Taiwan
Authors: Broadberry R. E. , Lin M.
Published: November 2003

The MiIII (Gp.Mur) phenotype is rare among Caucasians but has a much higher incidence among some Oriental populations. In Taiwan the population is relatively heterogenous and, as expected, the incidence of the MiIII phenotype was found to vary markedly among the different ethnic groups. The highest frequencies of the MiIII phenotype were found among some of the indigenous groups, namely the Ami, Yami and Puyuma groups, being 88.4%, 34.3% and 21.2%, respectively, which are also the highest frequencies among any population group reported so far. Antibody screening cells and antibody identification panels containing the MiIII phenotype can obviously be obtained very easily from Ami blood donors and compatible blood for patients with anti‐‘Mia’ can most easily be obtained from Chinese blood donors, except those living on the east coast.
SampLink: a new system for the collection of donor blood samples
Authors: Shinar E. , Michlin H.
Published: November 2003

Increasing demands for screening tests on blood donors, which require a large number of undiluted samples, and the rising concern for the safety of the phlebotomists have led to the development of a new blood sampling system. The new device was evaluated during the collection of 60 blood units, and compared with 50 control units collected using the ‘cut and drip’ method. The time required for both blood donation and donor samples collection, blood component quality, coagulation factors activation and haemolysis were studied. In addition, reports and recommendations of Magen David Adom phlebotomists were evaluated after collecting 75 000 units using the new device. Donor sample collection with the new device was comparable and somewhat shorter than with the ‘cut and drip’ method, while the blood unit collection time remained unchanged. There were no differences in plasma haemoglobin, factor VIII and platelet yields and morphology scores in blood units and donor samples collected by the two methods. There were no reported instances of needlesticks among phlebotomists using either method. The new device was simple to operate, improved blood donor samples collection practice and enabled the collection of as many undiluted donor samples as required for routine testing. In addition, the use of vacuum tubes ensured sterility, safety and standardization of samples. The components provided were comparable in quality to those prepared from units collected by other methods.
Comparison of two blood cell separators in collecting peripheral blood stem cell components
Authors: Stroncek D. F. , Clay M. E. , Smith J. , Jaszcz W. B. , Herr G. , McCullough J.
Published: November 2003

To ensure that a sufficient number of CD34+ cells are collected for an allogeneic blood progenitor cell transplant, the most effective blood cell separator should be used to collect peripheral blood stem cell (PBSC) components. We compared the effectiveness of two blood cell separators. We gave 29 healthy people 7.5 or 10 μg kg−1 of granulocyte colony stimulating factor (G‐CSF) daily for 5 days and collected one PBSC component with either a Fenwal CS3000 (n = 15) or a Cobe Spectra (n = 14) blood cell separator. The volume of blood processed was the same for each machine (8.4 ± 1.0 L; range = 4.9–9.4 L for the CS3000 and 8.9 ± 1.0 L; range 6.7–10.9 L; P = 0.71). The components collected with the CS3000 contained more mononuclear cells (39.6 ± 21.9 × 109 compared with 26.9 ± 5.6 × 109, P = 0.02) and fewer neutrophils (1.38 ± 1.88 × 109 compared with 5.53 ± 8.71 × 109,  = 0.001). The total number of CD34+ cells collected with the two instruments was the same (470 ± 353 × 106 for the CS3000 and 419 ± 351 × 106 for the Spectra; P = 0.64) as was the number of CD34+ cells collected per litre of whole blood processed (55.9 ± 42.0 × 106 L−1 compared with 45.9 ± 37.9 × 106 L−1; P = 0.59). The mononuclear cell collection efficiency was greater for the CS3000 (82.4 ± 54.9% compared with 53.3 ± 14.1; P = 0.04) but the CD34+ cell collection efficiencies were the same (87.4 ± 61.1% for the CS3000 compared with 56.3 ± 23.5% for the Spectra; P = 0.07). In conclusion, both blood cell separators collected components which contained large numbers of CD34+ cells, but those collected with the CS3000 contained fewer neutrophils and the CS3000 was more efficient at collecting mononuclear cells.
Blood spillage during salvage
Authors: Bengtson J. P. , Bengtsson A. , Rydberg L.
Published: November 2003

Recovery of blood components by salvage was evaluated in a randomized laboratory investigation using donor whole blood. Three methods of salvage were studied: conventional red cell salvage, red cell salvage with continuous processing, and salvage by haemofiltration using recirculation through a 30 000‐Da filter. Less than 10% of the haemoglobin was wasted during processing by all three methods. About half of the leukocytes were recovered with no significant difference between the techniques. The recovery of platelets was 15, 4 and 41% during salvage with conventional red cell centrifugation, continuous centrifugation processing and haemofiltration, respectively. In contrast with the other methods, haemofiltration gave a major recovery of albumin.
Effects on complement activation of a new continuous autotransfusion system
Authors: Bengtsson A. , Åvall A. , Tylman M. , Wilén G. , Bengtson J. P.
Published: November 2003

Allogeneic blood transfusions may subject patients to risks of infection and allergic reactions. Various techniques for transfusion of shed blood have been developed. The aim of this study was to evaluate a new continuous autotransfusion system (Fresenius CATS®) as regards its impact on the complement system, and on erythrocytes and leucocytes. Eighteen consecutive patients undergoing hip replacement surgery were studied. Complement variables (C4d, factor Bb, C3a and terminal complement complex, SC5b‐9) and free haemoglobin, haemoglobin, leucocytes, platelets, albumin and protein were determined in the patient’s blood preoperatively, 1 min before the start of transfusion, 15 and 60 min after transfusion; and in the reservoir, in the waste bag and in the retransfusion blood. Increased concentrations of C3a and SC5b‐9 were found in the collected reservoir blood (P < 0.05). The washing and centrifugation procedure reduced these concentrations (< 0.001). High levels of free haemoglobin were found in the collected blood as well as in the processed product. The median haemoglobin level in the processed blood was 260 g L−1 (range 104–289 g L−1). Inflammatory mediators from the complement cascade are removed by continuous autotransfusion technique. The processed blood contains high levels of free haemoglobin.
Length of survival after perioperative transfusion
Authors: Vamvakas E. C. , Moore S. B.
Published: November 2003

To describe the post‐transfusion survival of an entire, geographically defined population, we observed all residents of a US county who underwent perioperative transfusion before and after the introduction of a large autologous transfusion programme. We enrolled 444 and 1540 county residents, transfused in 1981 and in 1986–88, respectively. Complete follow‐up (until death or for 5 years) was available on 1960 patients (98.8%). Of patients transfused in 1986–88, 67.6% were alive at 5 years, having survived for a mean (± SE) period of 46.15 (±0.575) months. The survival statistics were 66.2% and 46.09 (±1.047) months, respectively, for patients transfused in 1981 (P =0.8424). Transfusion in 1986–88 vs. 1981 (with 615 [40%] vs. six [1.3%] patients receiving some autologous blood) did not have an effect on survival (P =0.3892), following adjustment for age, gender, transfusion dose, receipt of a single‐unit transfusion and transfusing surgical service. We conclude that the survival of an entire, geographically defined transfused population is substantially longer than that reported previously for patients referred to tertiary‐care medical centres. The aggregate 5‐year survival of patients transfused in 1981 and in 1986–88 does not differ, despite differences in patient case‐mix and in perioperative transfusion practice, particularly as it relates to autologous blood usage.
The presence of free infectious cytomegalovirus (CMV) in the plasma of donated CMV‐seropositive blood and platelets
Authors: James D. J. , Sikotra S. , Sivakumaran M. , Wood J. K. , Revill J. A. , Bullen V. , Myint S.
Published: November 2003

An in vitro study was carried out to determine the presence of free infectious CMV in the supernatant of donated units of blood and platelets which were known to be CMV seropositive. One sample was taken from each of 10 units of CMV‐seropositive platelet donations which were 4 days old. Ten units of seropositive blood were sampled at intervals over their storage lifespan. Each sample was divided into two and thrombin was added to one of them. The samples were all prepared by centrifugation. The resulting supernatant from the clotted samples was used in in vitro culture studies using the MRC‐5 cell line to ascertain the infectivity of the samples. The anticoagulated supernatants were subjected to PCR analysis to detect the presence of free genomic CMV DNA. All of the units of blood and platelets tested positive for the presence of genomic CMV DNA by PCR. Seven out of the 10 units of blood were culture positive from samples taken 3–4 weeks into their shelf‐life. None of the platelet samples was culture positive. The cultures of supernatants taken from seropositive blood were negative when the units were fresh, but further into their storage life, the cultures became positive, indicating that infectious material was released into the supernatant during storage, presumably due to the breakdown of intact white cells.
A comparison of monoclonal antibody immobilization of platelet antigen (MAIPA) and immunobead methods for detection of GPIIb/IIIa antiplatelet antibodies in immune thrombocytopenic purpura
Authors: Crossley A. , Calvert J. E. , Taylor P. R. A. , Dickinson A. M.
Published: November 2003

In this study we have compared two assays for the detection of autoantibodies GpIIb/IIIa, platelet bound and in serum, in immune thrombocytopenic purpura (ITP). Both assays were found to have a similar sensitivity, but the monoclonal antibody immobilization of platelet antigen (MAIPA) assay was more reproducible than the immunobead assay. The MAIPA and immunobead assay demonstrated an 81% concordance of results for serum antibody detection and a 78% concordance for platelet‐associated antibody detection, with an 8–12% incidence of false positive or negative results.
Comparison of Fyb status as determined serologically and genetically
Authors: Murphy M. T. , Templeton L. J. , Fleming J. , Ferguson M. , Peterkin M. , Fraser R. H.
Published: November 2003

Comparison of the serologically determined phenotype and the genotype of samples as defined by PCR‐RFLP indicated a significant percentage of discrepancies on analysis of serologically defined Fy(+b−) samples. An investigation of these discrepancies has shown that the serologically determined phenotype of the samples varies with respect to the reagent used but that the PCR‐RFLP assay gives reliable results for all the samples investigated. The samples which yielded the discrepant results were further investigated and the results indicated weakened expression of the Fyb antigen. This may arise by a method similar to the expression of Fyx although we found the incidence of these samples to be much higher than that reported for Fyx. These results may have important implications for the routine testing of blood donations, particularly in situations where the unit is to be transfused to a patient who has previously formed an anti‐Fyb.
The monoclonal antibody‐specific immobilization of erythrocyte antigens assay (MAIEA) in the investigation of human red‐cell antigens and their associated membrane proteins
Authors: Petty A. C. , Green C. A. , Daniels G. L.
Published: November 2003

The monoclonal antibody‐specific immobilization of erythrocyte antigens (MAIEA) technique is an immunoassay devised primarily for locating blood group antigens on specific red‐cell membrane proteins. The assay involves the incubation of intact red cells with two antibodies, one human alloantibody, the other a nonhuman antibody, usually a rodent monoclonal antibody, but polyclonal antibodies of rabbit origin have been utilized. For a positive result, both antibodies must bind to the same membrane protein. The red cells are lysed, the membrane solubilized and the trimolecular complex of two antibodies and membrane protein is captured in a well coated with goat antirodent (or rabbit) immunoglobulin. The immobilized complex is then detected by the use of peroxidase‐conjugated goat antihuman (or rodent) immunoglobulin.

Negative results, due to mutual blocking between the human and animal antibodies when their epitopes are close together on the same molecule, have permitted a degree of localization of epitopes on some proteins. This has been most effective in the mapping of Cromer blood group system antigens on the complement control protein domains of decay‐accelerating factor (DAF, CD55), but has also proved informative in the clustering of antigens on the Lutheran and Kell glycoproteins.

MAIEA is an effective tool for the identification of antibodies to Knops‐system antigens on complement receptor 1 (CR1, CD35) in immunohaematology reference laboratories. These antibodies are clinically unimportant, but must be identified before they can be ignored for transfusion purposes.
Cost analysis of autologous blood transfusion, using cell salvage, compared with allogeneic blood transfusion
Authors: Duffy G. , Tolley K.
Published: November 2003

Autologous transfusion is an option infrequently used in the UK, partly because the direct costs are greater than for allogeneic transfusion. An analysis of the cost consequences of substituting autologous for allogeneic blood, by using cell salvage, was undertaken from a hospital perspective. Direct costs were estimated for two different cell salvage devices and sensitivity analysis performed on the key variables.

Allogeneic transfusion may be associated with increased rates of postoperative infection due immunomodulation and immunosuppression. As a result, one of the short‐term benefits of autologous transfusion is a possible reduction in length of hospital stay. This was the most important variable affecting the cost of autologous transfusion. Cost equivalence for autologous and allogeneic blood was reached at reductions in hospital stay of between 0.3 and 2 days across a range of variables.

Reductions in length of hospital stay of this magnitude have been reported in several small studies. The results of our cost analysis suggest that autologous transfusion should not be rejected on the grounds of cost and we recommend a large‐scale randomized controlled trial of autologous vs. allogeneic transfusion.
Evaluation of awareness and utilization of an autologous blood transfusion programme
Authors: Dhingra‐Kumar N. , Sikka M. , Madan N. , Sood S. K.
Published: November 2003

Autologous blood transfusion (ABT) has an important role in transfusion practice in the developing world due to increasing incidence of HIV and hepatitis C virus infection. Our study was done to evaluate the level of awareness and utilization of an autologous blood transfusion programme in a teaching hospital in Delhi. We assessed the level of awareness of preoperative ABT amongst treating physicians from different specialties in a teaching hospital through an anonymous questionnaire. The utilization of this methodology in transfusion practice was estimated from records of the Blood Transfusion Service.

Of the 150 doctors contacted 96 (64%) responded. Although 67.7% of them were aware of the technique and its advantages, only 21.8% used it for the patients under their care. In the preceding 24 months 133 (1.1%) of 12 090 blood collections in the transfusion service were from autologous donor‐patients. Only one unit of blood was collected from each patient, although 41.8% of them received  2 units of blood. Of the 11 123 patients transfused, 55 (0.49%) received the ABT. Thus only 55 (41.3%) of 133 total ABT collections were utilized. The study highlights that there is a general lack of awareness about ABT amongst physicians. This transfusion practice is rarely and inadequately used. The study was repeated the following year after an intensive intervention strategy was adopted. The results show a trend towards improvement in the practice of ABT. This study emphasizes the need for proper organization, planning and communication between clinicians and blood transfusion personnel for effective implementation of an ABT programme, especially in countries with a high incidence of transfusion‐transmitted infections and acute shortages of blood for transfusion.
Citrate metabolism by human platelets
Authors: Cartledge S. , Candy D. J. , Hawker R. J.
Published: November 2003

As part of a study on the utilization of substrates by platelets in defined media the metabolism of citrate was measured, since citrate is a common anticoagulant of nearly all such media, and is also an intermediate of oxidative metabolism. Human platelets transferred from plasma to an artificial medium by gel filtration, were incubated with [14C]citrate at 22 °C and labelled carbon dioxide produced was measured during short‐term incubations of 2 h. Citrate (1 mm) was oxidized to carbon dioxide at low (0.3 nmol per 109 platelets h−1) but significant rates, and the oxidation was decreased by the presence of an alternative substrate (acetate) in the medium. There was, however, no significant conversion of citrate to glycogen. It was calculated that under normal storage conditions of platelet concentrates for transfusion purposes, the amount of citrate used cannot decrease citrate concentrations sufficiently to bring about platelet activation.
Process control of filtered red blood cells: which counting method?
Authors: Conte R. , Bontadini A. , Cirillo D. , Fruet F.
Published: November 2003

Various counting methods have been described and reported for process control of leucodepleted blood components. The recent production of high‐efficiency leucocyte removal filters intensifies the need for sensitivity in determining the ever lower residual concentration of white cells (WBCs) in filtered units.

In order to assess which method was the most efficient and feasible in the laboratory for the control of WBC‐reduced packed red blood cells, we compared the sensitivity of four counting methods: Nageotte chamber analysis, flow cytometry, the fluorochrome method by Borzini and Nageotte chamber analysis as modified by Prati.

We observed a difference in the post‐filtration WBC content depending on which method of counting was used and we feel it reasonable to ask what method should be employed in blood component process control.

The answer must naturally consider that the method is for use by a large number of laboratories, while the sensitivity of the method needs to be appropriate to the goal desired.
A quality assurance scheme for the Kleihauer test: the Scottish experience 1988–1996
Authors: Raafat A. , Fraser N. , Main R. , Urbaniak S. J.
Published: November 2003

This report describes the experience gained in the last 8 years of implementing and administering a Quality Assurance Scheme dedicated to improving the reliability of Kleihauer test results in Scotland. Initially the five transfusion centres in Scotland were the only participants, but since 1992 all 15 hospital laboratories in Scotland performing the Kleihauer test to ensure an adequate dose of anti‐D is administered to Rh‐negative women, at risk of developing allo anti‐D, have joined this voluntary scheme. As a result of the standardization of technical methods, improvement in reporting of QA results has been observed since 1994.
DBT: a partial D phenotype associated with the low‐incidence antigen Rh32
Authors: Wallace M. , Lomas‐Francis C. , Beckers E. , Bruce M. , Campbell G. , Chatfield S. , Nagao N. , Okubo Y. , Opalka A. , Overbeeke M. , Scott M. , Voak D.
Published: November 2003

Eight probands are described with a D phenotype in which a partial D antigen is associated with Rh32 antigen; three of these probands were investigated because of the anti‐D in their serum. The partial D lacks epD1–epD5 and epD9 and some epD6/7 and only expresses epD8 and other parts of epD6/7. The strength of the partial D antigen varies between unrelated DBT individuals. The Rh32 antigen of the DBT cells is weaker than that of D(C)(e) cells. Tests on DBT, DFR and R0Har cells with anti‐epD6/7 split these monoclonal anti‐D into eight patterns of reaction. A new pattern of reactions was observed, presaging a new epitope, but this was not numbered.
A mutation analysis on GYPC, the gene encoding the Gerbich blood group antigens
Authors: Johnson P. , Daniels G.
Published: November 2003

The four exons of GYPC genomic DNA were amplified together with their flanking sequences and analysed by denaturing gradient gel electrophoresis (DGGE) or single‐strand conformational analysis (SSCA). Exons 2 and 3 were amplified by a common set of primers so as to provide an internal control for exon deletion mutants. Ge:‐2,‐3,4 (Ge) and Ge:‐2,3,4 (Yus) phenotypes were always associated with exon 3 and exon 2 deletions, respectively, as determined by DGGE. Ge:‐2,‐3,‐4 (Leach) was associated with a deletion of exons 3 and 4. Two single base mutations within exon 2 were detected, one of which represented the Ana allele and encoded an amino acid substitution. A previously undetected single base mutation in exon 3 encoded a Val61Ile substitution within the membrane spanning domain. DGGE analysis of exon 4 product revealed two distinct haplotypes of frequencies 41% and 59%. DGGE was not found suitable for analysis of exon 1, so SSCA was utilized. This method distinguished the single base mutation responsible for Wb antigen, but not that responsible for Dha.