bluuebunny/crossref_metadata_2025 huggingface dataset:
A delayed leaf senescence mutant is defective in arginyl‐tRNA:protein arginyltransferase, a component of the N‐end rule pathway in Arabidopsis
Authors: Yoshida Satoko , Ito Masaki , Callis Judy , Nishida Ikuo , Watanabe Akira
Published: March 2003

We have isolated a delayed‐leaf‐senescence mutant, designated dls1, from an Arabidopsis T‐DNA line. Leaf senescence progresses more slowly in the dls1 mutant than in the wild‐type plant in both age‐dependent and dark‐induced senescence. Genetic analysis revealed that the dls1 is a monogenic recessive mutation that cosegregated with the T‐DNA insertion. Isolation of DNA flanking the T‐DNA revealed that the T‐DNA was inserted into the fourth intron of the AtATE1 gene, which encodes arginyl‐tRNA:protein arginyltransferase (EC. 2.3.2.8, R‐transferase), a component of the N‐end rule proteolytic pathway in yeast and mammals that transfers arginine to the N‐terminus of proteins with N‐terminal glutamyl or aspartyl residues. AtATE1 transcripts were not detectable in the dls1 mutant by RT‐PCR analysis. Introduction of a wild‐type AtATE1 gene into the dls1 mutant complemented the dls1 phenotype. We also showed using a transient expression assay system, that the dls1 mutation results in a decreased degradation of proteins with Asp or Glu at their N‐termini, and that the introduction of the wild‐type AtATE1 gene reverses this deficiency. These results suggest that the normal progression of leaf senescence requires R‐transferase activity, and that proteolysis by the N‐end rule pathway has an important physiological function in the progress of leaf senescence in plants.
COI1 links jasmonate signalling and fertility to the SCF ubiquitin–ligase complex in Arabidopsis
Authors: Devoto Alessandra , Nieto‐Rostro Manuela , Xie Daoxin , Ellis Christine , Harmston Rebecca , Patrick Elaine , Davis Jackie , Sherratt Leigh , Coleman Mark , Turner John G.
Published: March 2003

Jasmonates (JAs) regulate Arabidopsis thaliana wound and defence responses, pollen development, and stress‐related growth inhibition. Significantly, each of these responses requires COI1, an F‐box protein. Other F‐box proteins interact with SKP1 and cullin proteins to form SCF complexes that selectively recruit regulatory proteins targeted for ubiquitination. To determine whether COI1 also functions in an SCF complex, we have characterized Arabidopsis proteins that bind to COI1. An Arabidopsis cDNA expression library was screened in yeast for clones that produce proteins which can bind to COI1. We recovered two SKP1 homologues and a histone deacetylase. The Arabidopsis F‐box protein TIR1 interacted with SKP1 proteins, but not with the histone deacetylase. Mutant COI1 proteins revealed that the F‐box is required for interaction with SKP1s, but that sequences in leucine‐rich repeat domains are required for interaction with the histone deacetylase. Epitope‐tagged COI1 was introduced into Arabidopsis plants and cell cultures. Co‐immunoprecipitation experiments confirmed the interaction in planta of COI1 with SKP1‐like proteins and histone deacetylase, and also indicated that COI1 interacted with cullin. These results suggest that COI1 forms an SCFCOI1 complex in vivo. COI1 is therefore expected to form a functional E3‐type ubiquitin ligase in plants and to regulate expression of jasmonate responsive genes, possibly by targeted ubiquitination of a histone deacetylase.
RCY1, an Arabidopsis thaliana RPP8/HRT family resistance gene, conferring resistance to cucumber mosaic virus requires salicylic acid, ethylene and a novel signal transduction mechanism
Authors: Takahashi Hideki , Miller Jennifer , Nozaki Yukine ,   Sukamto, Takeda Megumi , Shah Jyoti , Hase Shu , Ikegami Masato , Ehara Yoshio , Dinesh‐Kumar S. P.
Published: March 2003

The dominant locus, RCY1, in the Arabidopsis thaliana ecotype C24 confers resistance to the yellow strain of cucumber mosaic virus (CMV‐Y). The RCY1 locus was mapped to a 150‐kb region on chromosome 5. Sequence comparison of this region from C24 and a CMV‐Y‐susceptible C24 mutant predicts that the RCY1 gene encodes a 104‐kDa CC‐NBS‐LRR‐type protein. The RCY1 gene from C24, when expressed in the susceptible ecotype Wassilewskija (Ws), restricted the systemic spread of virus. RCY1 is allelic to the resistance genes RPP8 from the ecotype Landsberg erecta and HRT from the ecotype Dijon‐17, which confer resistance to Peronospora parasitica biotype Emco5 and turnip crinkle virus (TCV), respectively. Examination of RCY1 plants defective in salicylic acid (SA), jasmonic acid (JA) and ethylene signaling revealed a requirement for SA and ethylene signaling in mounting a resistance response to CMV‐Y. The RCY1 nahG etr1 double mutants exhibited an intermediate level of susceptibility to CMV‐Y, compared to the resistant ecotype C24 and the susceptible ecotypes Columbia and Nossen. This suggests that in addition to SA and ethylene, a novel signaling mechanism is associated with the induction of resistance in CMV‐Y‐infected C24 plants. Moreover, our results suggest that the signaling pathways downstream of the RPP8, HRT, and RCY1 have evolved independently.
Tobacco plants were transformed by Agrobacterium rhizogenes infection during their evolution
Authors: Suzuki Kenji , Yamashita Ichiro , Tanaka Nobukazu
Published: March 2003

We discovered that the origin of cT‐DNA in the genome of wild‐type Nicotiana glauca is the T‐DNA of the mikimopine‐type Ri plasmid (pRi) harbored in Agrobacterium rhizogenes. The cT‐DNA was inserted into the genomic DNA of N. glauca from the position corresponding to the right border of mikimopine‐type pRi. The cT‐DNA contained two mikimopine synthase gene (mis) homologs, NgmisL and NgmisR, both of which were transcribed at low level in all N. glauca organs. NgMisR protein expressed in Escherichia coli has preserved Mis activity, which converts l‐histidine and α‐ketoglutaric acid to mikimopine. The mis homolog was also found in the genome of three other Nicotiana species: N. tomentosa, N. tomentosiformis, and N. tabacum; however, the site of insertion differed from that in N. glauca, suggesting that A. rhizogenes harboring mikimopine‐type pRi independently infected the ancestors of some Nicotiana plants. This is the first clear evidence of a host–parasite relationship during the early evolution of Nicotiana plants. We propose that a new phylogenetic approach using opine type cT‐DNA is applicable for presuming divergence in the genus Nicotiana.
Overexpression of a kinase‐deficient form of the EDR1 gene enhances powdery mildew resistance and ethylene‐induced senescence in Arabidopsis
Authors: Tang Dingzhong , Innes Roger W.
Published: March 2003

The EDR1 gene of Arabidopsis has previously been reported to encode a Raf‐like mitogen‐activated protein kinase kinase (MAPKK) kinase, and to function as a negative regulator of disease resistance. A phylogenetic analysis of plant and animal protein kinases revealed, however, that plant Raf‐like kinases are more closely related to animal mixed lineage kinases (MLKs) than Raf‐like kinases, and are deeply divergent from both classes of animal kinases, making inferences of substrate specificity questionable. We, therefore, assayed the kinase activity of recombinant EDR1 protein in vitro. The EDR1 kinase domain displayed autophosphorylation activity and phosphorylated the common MAP kinase substrate myelin basic protein. The EDR1 kinase domain also phosphorylated a kinase‐deficient EDR1 protein, indicating that EDR1 autophosphorylation can occur via an intermolecular mechanism. Overexpression of a kinase‐deficient full‐length EDR1 gene (35S::dnEDR1) in wild‐type Arabidopsis plants caused a dominant negative phenotype, conferring resistance to powdery mildew (Erysiphe cichoracearum) and enhancing ethylene‐induced senescence. RNA‐gel blot analyses showed that the 35S::dnEDR1 transgene was highly transcribed in transgenic plants. Western blot analysis, however, revealed that neither the wild‐type nor mutant EDR1 protein could be detected in these lines, indicating that the dominant negative phenotype may be caused by a translational inhibition mechanism rather than by a protein level effect. Overexpression of orthologous dnEDR1 constructs may provide a novel strategy for controlling powdery mildew disease in crops.
Susceptible to intolerance – a range of hormonal actions in a susceptible Arabidopsis pathogen response
Authors: O'Donnell Philip J. , Schmelz Eric A. , Moussatche Patricia , Lund Steven T. , Jones Jeffery B. , Klee Harry J.
Published: March 2003

Ethylene and salicylic acid (SA) are key intermediates in a host's response to pathogens. Previously, we have shown using a tomato compatible interaction that ethylene and SA act sequentially and are essential for disease symptom production. Here, we have examined the relationship between the two signals in the Arabidopsis–Xanthomonas campestris pv. campestris (Xcc) compatible interaction. Preventing SA accumulation by expression of the nahG gene reduced subsequent ethylene production and altered the development of disease symptoms, with plants showing no visible chlorosis. The ethylene insensitive lines, etr1‐1 and etr2‐1, on the other hand, accumulated SA and exhibited normal but precocious symptom development. Therefore, Arabidopsis, like tomato, was found to exhibit co‐operative ethylene and SA action for the production of disease symptoms. However, in Arabidopsis, SA was found to act upstream of ethylene. Jasmonic acid and indole‐3‐acetic acid levels were also found to increase in response to Xcc. In contrast to ethylene, accumulation of these hormones was not found to be dependent on SA action. These results indicate that the plants response to a virulent pathogen is a composite of multiple signaling pathways.
The GLOBULAR ARREST1 gene, which is involved in the biosynthesis of folates, is essential for embryogenesis in Arabidopsis thaliana
Authors: Ishikawa Takaaki , Machida Chiyoko , Yoshioka Yasushi , Kitano Hidemi , Machida Yasunori
Published: March 2003

We identified a mutation in Arabidopsis that resulted in defective embryos, and we designated this mutation globular arrest1 (gla1). The predicted amino acid sequence encoded by the GLA1 gene is homologous to the amino acid sequences of folylpolyglutamate synthetase (FPGS) and dihydrofolate synthetase (DHFS), which participate in folate biosynthesis. The defect of gla1 in the formation of calli was rescued by the supplement of 5‐formyl tetrahydrofolate. These results indicated that GLA1 is involved in the biosynthesis of tetrahydrofolate. The gla1 embryos developed normally in the early stage of development but did not undergo the transition to the heart stage. Thus, the function of the GLA1 gene in embryogenesis appears to be required after the globular stage. However, when the levels of GLA1 transcripts in transgenic plants were increased by introduction of several copies of a GLA1 transgene (GLA6.8), the gla1 embryos that grew on gla1/gla1 GLA6.8/– plants developed as far as the heart to bent‐cotyledon stage. This result suggests that the GLA1 function is provided to embryos by maternal tissues until embryos reach the globular stage.
GeBP, the first member of a new gene family in Arabidopsis, encodes a nuclear protein with DNA‐binding activity and is regulated by KNAT1
Authors: Curaba Julien , Herzog Michel , Vachon Gilles
Published: March 2003

Trichomes of Arabidopsis are single‐celled epidermal hair that are a useful model for studying plant cell fate determination. Trichome initiation requires the activity of the GLABROUS1 (GL1) gene whose expression in epidermal and trichome cells is dependent on the presence of a 3′‐cis‐regulatory element. Using a one‐hybrid screen, we have isolated a cDNA, which encodes for a protein, GL1 enhancer binding protein (GeBP), that binds this regulatory element in yeast and in vitro. GeBP and its three homologues in Arabidopsis share two regions: a central region with no known motifs and a C‐terminal region with a putative leucine‐zipper motif. We show that both regions are necessary for trans‐activation in yeast. A translational fusion with the Yellow Fluorescent Protein (YFP) indicates that GeBP is a nuclear protein whose localization is restricted to, on average, 3–5 subnuclear foci that might correspond to nucleoli. Transcriptional fusion with the GUS reporter indicates that GeBP is mainly expressed in vegetative meristematic tissues and in very young leaf primordia. We looked at GeBP expression in plants mutated in or misexpressing KNAT1, a KNOX gene, expressed in the shoot apical meristem and downregulated in leaf founder cells, and found that GeBP transcript level is regulated by KNAT1 suggesting that KNAT1 is a transcriptional activator of GeBP. This regulation suggests that GeBP is acting as a repressor of leaf cell fate.
Structure and expression of the rice class‐I type histone deacetylase genes OsHDAC1–3: OsHDAC1 overexpression in transgenic plants leads to increased growth rate and altered architecture
Authors: Jang In‐Cheol , Pahk Yoon‐Mok , Song Sang Ik , Kwon Ho Jeong , Nahm Baek Hie , Kim Ju‐Kon
Published: March 2003

Histone deacetylases (HDACs) modulate chromatin structure and transcription. HDACs have been studied as negative regulators in eukaryotic transcription. We isolated the rice OsHDAC1–3 genes for class I‐type histone deacetylases, which are related to the RPD3 family. The OsHDAC1 gene encoded a protein of approximately 57 kDa that shared 73.5, 72.7, 79.9, and 57.1% amino acid sequence identity with the OsHDAC2, OsHDAC3, maize RPD3, and human HDAC1 proteins, respectively. Genomic structures and Southern blot analyses revealed that OsHDAC1–3 contained seven, six, and seven exons, respectively, and constituted a class I‐type family in the rice genome. OsHDAC1 was expressed at similar levels in the leaves, roots, and callus cells, whereas OsHDAC2 and 3 were expressed in the roots and callus cells, but not in the leaves, exhibiting distinct tissue specificity. To explore the role of histone deacetylases in transgenic plants, we inserted the OsHDAC1 cDNA fragment into the expression vector Ai::OsHDAC1 under the control of the ABA‐inducible promoter Ai, and transformed the construct into rice. Levels of mRNA, protein, and HDAC activity were significantly increased in Ai::OsHDAC1 callus cells. The amount of tetra‐acetylated H4 in the transgenic cells was greatly reduced, and the reduction was abolished upon treatment with trichostatin A. These results demonstrate that OsHDAC1 overexpression in transgenic cells both yields enzymatically active HDAC complexes and induces changes in histone acetylation in vivo. The overexpression leads to a range of novel phenotypes, involving increased growth rate and altered plant architecture, suggesting that OsHDAC1 functions in the genome‐wide programming of gene expression.
Interactions between gene activity and cell layers during floral development
Authors: Vincent Coral A. , Carpenter Rosemary , Coen Enrico S.
Published: March 2003

The DEFICIENS (DEF) gene is required for establishing petal and stamen identity in Antirrhinum and is expressed in all three layers of the floral meristem in whorls 2 and 3. Expression of DEF in a subset of meristem layers gives rise to organs with characteristic shapes and cell types, reflecting altered patterns and levels of DEF gene activity. To determine how the contributions of layers and gene activity interact, we exploited a DEF allele which carries a transposon insertion in the MADS box region to generate periclinal chimeras expressing alleles with different activities. By comparing the phenotype, development and expression patterns of these chimeras we show that expression of DEF in L1 makes a major contribution to morphology in whorl 2, irrespective of the allele. By contrast L1 expression is largely unable to rescue whorl 3, possibly because of a non‐autonomous inhibitor of DEF activity in this whorl.
Two means of transcriptional reactivation within heterochromatin
Authors: Probst Aline V. , Fransz Paul F. , Paszkowski Jerzy , Scheid Ortrun Mittelsten
Published: March 2003

DNA methylation levels and specific histone modifications of chromatin in interphase nuclei are taken as an indicator of transcriptional activity or silencing. Arabidopsis mutants impaired in maintenance of transcriptional gene silencing (TGS) alleviate TGS with or without affecting DNA methylation. Mutant ddm1, representing the first type, lacks a chromatin remodeling factor that regulates histone and DNA methylation. Mutant mom1, representing the second type, is affected in a different but still unknown silencing mechanism. Both classes of mutation have been studied mainly for their effects on specific loci. Here, we describe the cytological analysis of chromatin in ddm1 and mom1 mutants. The ddm1 mutation causes a striking decondensation of centromeric heterochromatin, a re‐distribution of the remaining methylation of DNA, and a drastic change in the pattern of histone modification. A complex transgenic locus, which underwent stable inactivation and became heterochromatin‐like, follows similar structural alterations. In contrast, nuclear organization in mom1 appears unaltered, demonstrating an involvement of MOM1 in transcriptional regulation within a heterochromatic environment.
The swirling phenomenon in stored platelets is influenced by their endogenous serotonin
Authors: Hervig  , Bakken  , Farstad
Published: March 2003

The presence of the swirling phenomenon is useful to define platelet concentrates that are suitable for transfusion. If it is possible to identify donor‐related factors which are related to persisting swirling during storage, it is possible to select platelet donors. Endogenous platelet serotonin content is stable and easily measured and related to agonist‐induced serotonin secretion.

During a 3‐month period, the swirling in 825 single donor platelet concentrates was controlled before issue. Endogenous serotonin, % serotonin release and swirling were tested in 21 concentrates with poor or no swirling during storage. Sixty‐three concentrates were randomly selected from the routinely prepared platelet concentrates and were routinely tested with the same analyses on days 1 and 7. To evaluate an obvious effect of endogenous serotonin on the swirling phenomenon, eight platelet concentrates prepared from buffy coat, each from four donors, were divided. One part was stored in the presence of 8.5 μmol serotonin L−1, and analysed as the control concentrates.

The endogenous serotonin content in the ‘low‐ swirling’ concentrates was significantly lower than in the control group (P < 0.001). P
co2 and pH were significantly lower, and P
o2 and MPV significantly higher than in the controls. In the control group, swirling after 7 days was significantly correlated with serotonin release. In the eight buffy‐coat concentrates enriched in endogenous serotonin, both swirling and the percentage serotonin release were improved after storage for 10 days, compared with nonenriched concentrates.

This study suggests that endogenous serotonin content and serotonin release are factors that may be of significance concerning preservation of the swirling phenomenon in platelet concentrates during storage.
A home blood transfusion programme for β‐thalassaemia patients
Authors: Madgwick  , Yardumian
Published: March 2003

Although home transfusion programmes are relatively common in the USA ( Anon, 1990, Home Care in the 1990s. Council on Scientific Affairs. Journal of the American Medical Association, 263, 1241–1244), this type of treatment has only recently been considered in Britain and, where it is in operation, is generally supervised by trained nursing staff or via a hospice. North Middlesex Hospital now has 3 years experience of a home transfusion programme operating for β‐thalassaemia major patients, in which relatives are trained and responsible for supervision of the red‐cell transfusions at home. For families who request this service, and who are willing and able to undertake it responsibly, the scheme offers the advantages of improved patient comfort, reduced absences from education or employment and reduced hospital bed usage. Patients and their carers express improved satisfaction with the treatment delivered in this way.
In vitro characteristics of red blood cell concentrates prepared from under‐ and overcollected units of whole blood and from a paediatric blood bag system
Authors: Weisbach  , Hunold  , Zimmermann  , Lutter  , Parsch  , Zingsem  , Glaser  , Eckstein
Published: March 2003

The problem of how to deal with red blood cell concentrates (RBCs) prepared from under‐ or overcollected units of whole blood (WB) and how to collect blood from underweight persons arises in the context of autologous predeposit. To determine the quality of RBCs stored in PAGGS‐M additive solution prepared from under‐ and overcollected units of whole blood and of PAGGS‐M RBCs prepared from a paediatric 250‐mL top outlet blood bag system we measured blood picture, haemolysis, K+, pH, ATP and 2,3‐DPG on days 0, 10, 20, 30, 40 and 49 of storage. The volume of WB collected ranged from 150 to 600 mL in 50‐mL increments (4 units per volume). Haemolysis was under 0.8% on day 49 in all RBCs prepared from WB donations between 200 mL and 600 mL. However, the day 49 haemolysis level of standard RBCs prepared from 450 mL of WB (0.15 ± 0.03%) was reached earlier in RBCs from under‐ and overcollected units of whole blood. 2,3‐DPG levels decreased rapidly between days 10 and 20 in all RBCs studied. RBCs from 450‐mL donations showed acceptable ATP maintenance after 49 days (70.4% of day 0 value), while all other RBC ATP levels were below 50% of the day 0 level on day 49. In vitro quality data of RBCs prepared from a 250‐mL donation in the paediatric blood bag system after storage for about 25 days were comparable to those after 49 days of storage of standard RBCs. Our results suggest that it is feasible to transfuse PAGGS‐M RBCs prepared from under‐ as well as overcollected units of WB in the autologous setting. However, we strongly recommend shortening the storage period of such RBCs to maintain the quality level of standard RBCs.
Seroprevalence of hepatitis B surface antigen (HBsAg) among first‐time and sporadic blood donors in Greece: 1991–1996
Authors: Kyriakis  , Foudoulaki  , Papoulia  , Sofroniadou
Published: March 2003

Hepatitis B surface antigen (HBsAg) seroprevalence among three major groups of sporadic voluntary blood donors in Greece was studied and compared to the seroprevalence in regular donors. These three groups share many characteristics with the general population. A 6‐year retrospective seroepidemiological study was carried out (1991–1996). The study population consisted of donors who were (i) military recruits (n = 80 302), (ii) enlisted military personnel (n = 86 920) and (iii) directed family donors (n = 75 403). A specimen was considered as HBsAg positive when found repeatedly reactive by a 3rd‐generation immunoassay and confirmed by RIA. The Mantel–Haenszel χ2 procedure was used for stratified analysis of the prevalence rates and Greenland/Robins confidence intervals of the respective weighted relative risk (MHRR) were calculated. The 6‐year overall HBsAg seroprevalence among the three sporadic donor groups was 0.84%; this was twice the seroprevalence among a sample of regular donors (n = 45 504) in Greece. Seroprevalence was higher among enlisted personnel (1.21 < MHRR = 1.34 < 1.50), during years prior to 1995. Directed family donors had the same overall seropositivity rate as recruits and enlisted personnel. After 1995, all groups had a seroprevalence below 1%, possibly indicating a shift towards lower endemicity in the Greek population.
How to maintain blood supply during computer network breakdown: a manual backup system
Authors: Zeiler T. , Slonka J. , Bürgi H. R. , Kretschmer V.
Published: March 2003

. Electronic data management systems using computer network systems and client/server architecture are increasingly used in laboratories and transfusion services. Severe problems arise if there is no network access to the database server and critical functions are not available. We describe a manual backup system (MBS) developed to maintain the delivery of blood products to patients in a hospital transfusion service in case of a computer network breakdown.

All data are kept on a central SQL database connected to peripheral workstations in a local area network (LAN). Request entry from wards is performed via machine‐readable request forms containing self‐adhesive specimen labels with barcodes for test tubes. Data entry occurs on‐line by bidirectional automated systems or off‐line manually. One of the workstations in the laboratory contains a second SQL database which is frequently and incrementally updated. This workstation is run as a stand‐alone, read‐only database if the central SQL database is not available. In case of a network breakdown, the time‐graded MBS is launched. Patient data, requesting ward and ordered tests/requests, are photocopied through a template from the request forms on special MBS worksheets serving as laboratory journal for manual processing and result report (a copy is left in the laboratory). As soon as the network is running again the data from the off‐line period are entered into the primary SQL server.

The MBS was successfully used at several occasions. The documentation of a 90‐min breakdown period is presented in detail. Additional work resulted from the copy work and the belated manual data entry after restoration of the system. There was no delay in issue of blood products or result reporting.

The backup system described has been proven to be simple, quick and safe to maintain urgent blood supply and distribution of laboratory results in case of unexpected network breakdown.
Utilization of technologies to reduce allogeneic blood transfusion in the United States
Authors: Hutchinson A. B. , Fergusson D. , Graham I. D. , Laupacis A. , Herrin J. , Hillyer C. D.
Published: March 2003

Concern over safety of the blood supply has led to the use of technologies to reduce allogeneic blood transfusion. The objective of this research was to determine the utilization of these technologies in the United States. We evaluated the following techniques: preoperative autologous donation (PAD), cell salvage (CS) and acute normovolemic haemodilution (ANH); and the following pharmaceuticals: aprotinin (APR), epsilon‐aminocaproic acid (EACA), tranexamic acid (TXA), desmopressin (DDAVP) and recombinant human erythropoietin (EPO). In 1997, we conducted a cross‐sectional mail survey of service chiefs at 1000 US hospitals randomly selected and stratified by status as a provider of open‐heart surgery, geographical location and hospital bed size. Sixty‐nine per cent (690) of hospitals responded to at least one of the four surveys sent to each hospital. Hospitals reported use of techniques more than pharmaceuticals (P < 0·001); PAD (83%, n = 206) and CS (82% n = 420) were used most frequently. Lack of familiarity was the most common reason cited for infrequent use of pharmaceuticals. Organizational characteristics (e.g. provision of open‐heart surgery, size, geographical location, teaching status and type of hospital) were differentially associated with technology use. There is greater use of techniques than pharmaceuticals in US hospitals to reduce the need for allogeneic blood in the peri‐operative setting. Providing open‐heart surgery is strongly associated with the utilization of these technologies.
HTLV antibody screening using mini‐pools
Authors: Dow B. C. , Munro H. , Ferguson K. , Buchanan I. , Jarvis L. , Jordan T. , Franklin I. M. , McClelland M.
Published: March 2003

. At the present time, the UK blood transfusion services do not screen blood donations for anti‐HTLV. This presentation describes a pilot study to ascertain the feasibility of HTLV antibody screening using mini‐pools and also provides an estimate of HTLV prevalence within our donor population in Scotland and Northern Ireland. The Abbott/Murex HTLV I/II GE80/81 ELISA was selected for the trial. Thirty confirmed HTLV positive library samples were tested at various dilutions and five were shown to be nonreactive at a dilution of 1 : 100. Residues of mini‐pools (of up to 95 individual donations) prepared for HCV NAT testing were tested with the Abbott/Murex GE80/81 assay. Of 6666 mini‐pools (equivalent to 570 609 donations) tested, six were repeatedly reactive. All six mini‐pools were confirmed HTLV antibody positive by line immunoassay. Four were confirmed to be HTLV‐I positive, one HTLV‐II positive and one HTLV positive (unable to type). Dilutions (1 : 100) of the five HTLV ‘nonreactive’ positive samples were included in each test plate and used to determine a grey‐zone cut‐off. Using this grey‐zone system an additional six (0·09%) mini‐pool samples gave repeatedly reactive grey‐zone results, none of which were confirmed. The minimum Scottish/Irish HTLV donor prevalence was shown to be 1 : 95 000.
Platelet chemokines and their receptors: what is their relevance to platelet storage and transfusion practice?
Authors: Boehlen F. , Clemetson K. J.
Published: March 2003

. The role of platelets as inflammatory cells is demonstrated by the fact that they can release many growth factors and inflammatory mediators, including chemokines, when they are activated. The best known platelet chemokine family members are platelet factor 4 (PF4) and β‐thromboglobulin (β‐TG), which are synthesized in megakaryocytes, stored as preformed proteins in α‐granules and released from activated platelets. However, platelets also contain many other chemokines such as interleukin‐8 (IL‐8), growth‐regulating oncogene‐α (GRO‐α), epithelial neutrophil‐activating protein 78 (ENA‐78), regulated on activation normal T expressed and secreted (RANTES), macrophage inflammatory protein‐1α (MIP‐1α), and monocyte chemotactic protein‐3 (MCP‐3). They also express chemokine receptors such as CCR4, CXCR4, CCR1 and CCR3. Platelet activation is a feature of many inflammatory diseases such as heparin‐induced thrombocytopenia, acquired immunodeficiency syndrome, and congestive heart failure. Substantial amounts of PF4, β‐TG and RANTES are released from platelets on activation, which may occur during storage. Although very few data are available on the in vivo effects of transfused chemokines, it has been suggested that the high incidence of adverse reactions often observed after platelet transfusions may be attributed to the chemokines present in the plasma of stored platelet concentrates.
A pilot study to assess physician knowledge in transfusion medicine
Authors: Rock G. , Berger R. , Pinkerton P. , Fernandes B.
Published: March 2003

Summary Introduction An assessment of transfusion medicine knowledge and practice in Canada was carried out over a 3‐week period.

Methods Fifty‐five questions were posed to evaluate both basic information on blood and blood products and on clinical use. The form was distributed through the Canadian Society for Transfusion Medicine with designated individuals asked to handle regional distribution. Some used provincial mailing lists, others distributed within each hospital. Approximately, 2000 forms were sent, including 500 in French.

Results A total of 294 forms were returned; answers were recorded as ‘correct’, ‘incorrect’, ‘no answer’ or with ‘added comments’. Overall, 52% of the questions were answered correctly or were answered with qualifying comments. In clinical practice questions, 63% were answered correctly or with qualifying comments. Basic knowledge questions drew correct answers in 37% of the cases. Several issues were answered poorly. The volume of an apheresis plasma unit was correctly estimated by less than 10% of respondents with many understating the volume (500–600 mL) by as much as 300–400 mL. Anaesthesiologists responded most often (21%); few haematologists participated (4%). Provincial response varied: most were received from Ontario (30%) and British Columbia (22%).

Conclusions The answers show that clinical application of transfusion is generally accompanied by a questioning process – it is not entirely by rote. Basic knowledge about products needs improvement.
Relative versus absolute risk of dying reduction after using insecticide‐treated nets for malaria control in Africa
Authors: Lengeler Christian , Armstrong‐Schellenberg Joanna , D'Alessandro Umberto , Binka Fred , Cattani Jacqueline
Published: March 2003

Four recently completed large‐scale randomized controlled trials have assessed the impact of insecticide‐treated bednets and curtains on overall child mortality in Africa. These results have sparked numerous discussions among implementing agency representatives and researchers about the public health significance of the results. For the interpretation of impact, most of the arguments have been based on the observed decrease in protective efficacy (a relative measure of impact) with increasing malaria transmission (range: 14–29%, regression for trend: F = 245 on 1,2 DF, P < 0.003). However, an analysis of the absolute measure of impact (the risk difference) showed a different pattern. The impact ranges from 3.8 to 6.9 lives saved per 1000 children protected per year, without a significant trend (F = 2.8 on 1,2 DF, P = 0.2) and with equally high values in both low and high transmission sites. When assessing the public health importance of an intervention, both relative and absolute decrease in risk should be considered.
Is apoptosis involved in mechanisms to eliminate Onchocerca ochengi during Simulium damnosum s.l. immune response?
Authors: Hagen Hans‐E. , Sabine L. Kläger  , Williams Gwyn T.
Published: March 2003

Co‐injection of the parasite Onchocerca ochengi and the caspase inhibitors z‐VAD.fmk and boc‐D.fmk into the natural vector Simulium damnosum s.l. led to significantly increased survival of the parasites. Subsequent in situ apoptosis detection assays demonstrated that in the case of boc‐D.fmk the enhanced survival was due to a diminished apoptosis level of the microfilariae in vivo. Additional assays using O. ochengi microfilariae which were coinjected with serine protease inhibitors into S. damnosum s.l. revealed that certain serine protease inhibitors can reduce the level of apoptosis.
Migration and health in the European Union
Authors: Carballo Manuel , Divino Jose Julio , Zeric Damir
Published: March 2003

The paper gives a brief overview of a wide spectrum of health issues and problems, ranging from communicable disease to mental health and family formation, which affect migrants and host countries.
Economics of theileriosis control in Zambia
Authors: D'haese L. , Penne K. , Elyn R.
Published: March 2003

Summary

For an economic analysis of theileriosis control, we adopted the total economic cost (TEC) method, which calculates the sum of output losses from tick damage, theileriosis mortality and morbidity, and expenditures for treatment or prevention of the disease. At farm level, the TEC can be minimized by a specific combination of vector control and/or immunization and an acceptable level of losses. Expenditures for vector control include acaricides, construction of dipping or spraying facilities and their maintenance, and variable costs such as those for water and labour. Economics of vector control depend on the herd size and the method of application of the acaricide. Morbidity, mortality and tick damage losses are effectively reduced by correct and intensive vector control programmes. Expenditures for vector control are estimated at US$ 8.43, 13.62 and 21.09 per animal per year for plunge dipping, hand spraying and pour‐on, respectively. Immunization costs comprise production of parasite stabilates, storage and application, delivery and treatment. At US$ 9.5 per animal, immunization limits losses caused by Theileria parva, but ticks still may reduce the productivity of the animals. Expenditures for treatment after natural infection involve drugs, transport, veterinary fees and farm labour costs. Treatment has a moderate success rate, hence both morbidity and mortality remain important factors. Equally, it does not affect the vector, which may continue to reduce overall productivity of cattle. Expenditures for treatment range between US$ 9.04 and US$ 27.31 per animal. To compare different TECs in relation to different control strategies, assumptions have to be made on disease occurrence, case fatality, value and productivity of the cattle, reductions in productivity due to morbidity and number of animals under a specific control regime. Calculations based on data from Southern Province, Zambia show that large‐scale immunization reduces the TEC by 90% compared to no intervention. Treatment, which is the second‐best option, reduces the TEC by 60%.

Appendix 1 Summary of factors influencing total economic cost
Dry supplementary feeding programmes: an effective short‐term strategy in food crisis situations
Authors: Vautier F. , Hildebrand K. , Dedeurwaeder M. , Van Herp M.
Published: March 2003

Summary Malnutrition is frequently a predominant problem in disasters, and supplementary feeding programmes (SFPs) are often set up in food emergencies. This review analyses the effectiveness of such programmes in crisis situations in Liberia, Burundi and Goma (Congo), concluding that it is feasible to enrol large numbers of children in SFPs and achieve proportions of recovery above 75% if these programmes are implemented as a short‐term measure in emergency situations. However, satisfactory SFP results do not necessarily indicate improved nutritional status of the whole population.
Diversité du régime alimentaire de Glossina palpalis palpalis en zone forestière de Côte d'Ivoire: relation avec la prévalence de la trypanosomiase humaine africaine
Authors: Sané Bocar , Laveissière Claude , Meda Honoré A.
Published: March 2003

RésuméUne analyse rétrospective des préférences trophiques de Glossina palpalis palpalis, vecteur majeur de la Trypanosomiase humaine africaine (THA) a été faite à partir des données recueillies entre 1984 et 1994 dans cinq foyers du secteur forestier, dans le centre‐ouest de la Côte d'Ivoire. Les auteurs ont comparé le régime alimentaire de ce vecteur dans ces différents foyers. L'objectif est de vérifier s'il existe une relation entre les préférences trophiques du vecteur et la prévalence de la THA. La diversité du régime alimentaire a étéévaluée par deux indices: l'indice de diversité de Shannon et Weaver (Ish) et un nouvel indice défini par le rapport pourcentage de repas de sang animal/pourcentage de repas de sang humain ou indice de zoophilie/ anthropophilie (Za). Il n'y a pas de correlation entre la DAP et le taux de prévalence. Des indices (Ish et Za) élevés, signe d'une diversité alimentaire, ont été observés dans les foyers de Vavoua, Zoukougbeu et Sinfra où les taux de prévalence de la THA étaient importants. Inversement des indices faibles ont été notés dans les zones de faible prévalence comme Daniafla et Gagnoa où la quasi totalité des repas de sang du vecteur était pris sur l'hôte humain. Il existe une corrélation forte mais non significative entre les deux indices entomologiques et la prévalence de la maladie. La corrélation est plus forte avec l'indice Za qu'avec l'indice Ish. La signification épidémiologique de ces observations est discutée.

Summary The feeding habits of Glossina palpalis palpalis, the main vector of human African trypanosomiasis (HAT) were retrospectively analysed using data collected between 1984 and 1994 in five areas in the forest belt in the mid‐west of Côte d'Ivoire. The authors compare the feeding habits of the vector in these different foci. This analysis is aimed at determining if there is any relationship between the feeding pattern of tsetse‐flies and the prevalence rates of HAT. The feeding pattern was measured using two indices: the conventional index of Shannon and Weaver (Ish) and a new one, the zoophily/anthropophily index (Za). The latter is an estimate of the ratio of the percentage of animal blood meals divided by the percentage of human blood meals. There was no correlation between apparent density and prevalence rate. A high Ish and a high Za were observed in the foci of Vavoua, Zoukougbeu and Sinfra where prevalence rates of HAT were high. Conversely, a low Ish and a low Za were observed in the hypoendemic areas of Daniafla and Gagnoa. Both indices are highly but not significantly correlated with prevalence rates. The Za index seemed to be more strongly correlated to the disease rate as compared to the Ish index. The epidemiological significance of these observations is discussed.
An open comparative study of dispersible piroxicam versus soluble acetylsalicylic acid for the treatment of osteoarticular painful attack during sickle cell crisis
Authors: Eke Felicia U. , Obamyonyi A. , Eke N. Ndubuisi , Oyewo E. A.
Published: March 2003

Summary We compared the efficacy and tolerability of oral piroxicam 1 mg/kg/day with soluble aspirin given at 100 mg/kg/day taken four‐hourly in 58 patients with sickle cell anaemia and severe ostcoarticular painful attacks requiring hospitalization in a randomized, paralleled study. Main investigational criteria were pain relief, limitation of movement, fever, and insomnia or agitation. Both groups were well‐matched at the commencement of therapy but most patients on piroxicam showed remarkable and significant pain relief and improvement in other parameters within 24 h. Unwanted effects were absent in the piroxicam‐treated group whereas those treated with aspirin experienced nausea and vomiting. There were no significant changes in liver function tests with both forms of treatment. Oral piroxicam is an effective and safe treatment in the management of the osteoarticular painful crisis in sickle cell anaemia. It might prevent the use of parenteral analgesics and hospitalization and reduce the loss of school hours in patients who are being treated for bone pain crises that characterize sickle cell anaemia.
Paediatric survival and re‐admission risks following hospitalization on the Kenyan Coast
Authors: Snow R. W. , Howard S. C. , Mung'ala‐Odera V. , English M. , Molyneux C. S. , Waruiru C. , Mwangi I. , Roberts D. J. , Donnelly C. A. , Marsh K.
Published: March 2003

Summary The district general hospital (DGH) is a common feature of health service provision in many developing countries. We have used linked demographic and clinical surveillance in a rural community located close to a DGH on the Kenyan coast to define the use and public health significance of essential clinical services provided by it. Of a birth cohort of over 4000 children followed for approximately 6 years, about a third were admitted to hospital at least once. Significantly more children admitted with major infectious diseases such as malaria and acute respiratory tract infections were readmitted with the same condition during the surveillance period than would have been expected by chance. Among surviving admissions, mortality post‐discharge was significantly higher than in the cohort which had not been admitted within 3, 6 and 12 months. Most of the patients who died after discharge had been admitted with a diagnosis of gastroenteritis. Most children admitted to the DGH survive hospitalization and the remaining period of childhood. Despite no clinical trial evidence to support the claim, it seems reasonable to assume that in the absence of intensive clinical management provided by a DGH, a significant proportion of these children would not have survived. However, the DGH is able to define a group of at‐risk children who re‐present with severe complications of infectious disease, and of these several may have underlying conditions not amenable to DGH intervention and continue to have a poor prognosis. Both groups of children represent statistically significant subsets of a rural paediatric community and the future organization and co‐ordination of DGH and primary care services need to work in unison to strengthen the service needs of children at risk.
In vivoefficacy study of amodiaquine and sulfadoxine/ pyrimethamine in Kibwezi, Kenya and Kigoma, Tanzania
Authors: Gorissen Erwin , Ashruf Gretchen , Lamboo Monique , Bennebroek Jacqueline , Gikunda Stephen , Mbaruku Godfrey , Kager Piet A.
Published: March 2003

SummaryWe conducted two randomized clinical trials to determine thein vivoefficacy of amodiaquine and sulfadoxine/pyrimethamine in treatingPlasmodium falciparummalaria. Seventy‐five patients under the age of 10 years in Kibwezi, Kenya, and 171 patients in Kigoma, Tanzania, were enrolled for treatment. Due to loss of eight patients in Kibwezi and 37 in Kigoma to follow‐up, we used best and worst case scenarios for the parasitological response. Thein vivosensitivity ofPlasmodium falciparumto amodiaquine was 75% (no loss to follow‐up) in Kibwezi and ranged from 85% in the best to 65% in the worst case scenario in Kigoma. The sensitivity to sulfadoxine/pyrimethamine was 70% to 88% in Kibwezi and 65% to 89% in Kigoma. R1 resistance to amodiaquine was 22% in Kibwezi and varied from 6% in the best to 26% for the worst case scenario in Kigoma. The R1 resistance to sulfadoxine/pyrimethamine was 5% to 23% in Kibwezi and 2% to 26% in Kigoma. R2 resistance was 3% for amodiaquine and 7% for sulfadoxine/pyrimethamine in Kibwezi and 9% in Kigoma for each treatment group. There was no statistically significant difference between treatment groups at either study site, except for a slight difference in R1 resistance in the best case scenario, Kibwezi, in favour of S/P. Although both amodiaquine and sulfadoxine/pyrimethamine resistance seems to be increasing, these antimalarials are still effective in parasite clearance.
Indoor residual spraying with alphacypermethrin controls malaria in Pakistan: a community‐randomized trial
Authors: Rowland Mark , Mahmood Pervaiz , Iqbal Javed , Carneiro Ilona , Chavasse Desmond
Published: March 2003

Summary We conducted a community‐randomized controlled trial in Pakistan to determine the efficacy of indoor residual spraying with alphacypermethrin ('Fendona', Cyanamid, NJ, USA), applied at 25 mg/m2, to prevent falciparum and vivax malaria. Wettable powder (WP) and suspension concentrate (SC) formulations were tested against an unsprayed control in an irrigated rice‐growing area of Sheikhupura district, Punjab province. The study area of 180 km2 was divided into nine sectors, which were assigned at random to control, WP, or SC treatments in replicates of 3. Sentinel villages were selected from each sector for entomological and disease monitoring. Malaria was monitored by fortnightly active case detection (ACD) and by cross‐ sectional parasite surveys on schoolchildren. Mosquito populations were monitored by space spraying of rooms and by cattle‐landing catches. The spray campaign took place in June 1997 and covered 96% of compounds. During the 7 months after spraying, the incidence of falciparum malaria was 95% lower and that of vivax malaria 80% lower in WP‐sprayed than unsprayed sectors. Similar results were obtained for sectors sprayed with the SC formulation. Cross‐sectional surveys gave estimates of efficacy comparable to those obtained by ACD. Anopheles culicifacies was 80% less abundant and A.stephensi, the predominant anopheline, was up to 68% less abundant in sprayed areas over the 7‐month period. Reductions in anopheline parous rates indicated that the single‐spray treatment was effective for the entire transmission season. Sprayed surfaces lacked odour, which contributed to the popularity of the campaign. Alphacypermethrin is a promising insecticide for the control of malaria in Pakistan and South Asia generally.