bluuebunny/crossref_metadata_2025 huggingface dataset:
STAIRS: a new genetic resource for functional genomic studies of Arabidopsis
Authors: Koumproglou Rachil , Wilkes Tim M. , Townson Paul , Wang Xiao Y , Beynon Jim , Pooni Harpal S. , Newbury H. John , Kearsey Mike J.
Published: March 2003

Many biologically and economically important traits in plants and animals are quantitative/multifactorial, being controlled by several quantitative trait loci (QTL). QTL are difficult to locate accurately by conventional methods using molecular markers in segregating populations, particularly for traits of low heritability or for QTL with small effects. In order to resolve this, large (often unrealistically large) populations are required. In this paper we present an alternative approach using a specially developed resource of lines that facilitate QTL location first to a particular chromosome, then to successively smaller regions within a chromosome (≤ 0.5 cM) by means of simple comparisons among a few lines. This resource consists of ‘Stepped Aligned Inbred Recombinant Strains’ (STAIRS) plus single whole Chromosome Substitution Strains (CSSs). We explain the analytical power of STAIRS and illustrate their construction and use with Arabidopsis thaliana, although the principles could be applied to many organisms. We were able to locate flowering QTL at the top of chromosome 3 known to contain several potential candidate genes.
Opposite changes in membrane fluidity mimic cold and heat stress activation of distinct plant MAP kinase pathways
Authors: Sangwan Veena , Örvar Björn Lárus , Beyerly John , Hirt Heribert , Dhindsa Rajinder S.
Published: March 2003

Mitogen‐activated protein kinases (MAPKs) appear to be ubiquitously involved in signal transduction during eukaryotic responses to extracellular stimuli. In plants, no heat shock‐activated MAPK has so far been reported. Also, whereas cold activates specific plant MAPKs such as alfalfa SAMK, mechanisms of such activation are unknown. Here, we report a heat shock‐activated MAPK (HAMK) immunologically related to ERK (Extracellular signal‐Regulated Kinase) superfamily of protein kinases. Molecular mechanisms of heat‐activation of HAMK and cold‐activation of SAMK were investigated. We show that cold‐activation of SAMK requires membrane rigidification, whereas heat‐activation of HAMK occurs through membrane fluidization. The temperature stress‐ and membrane structure‐dependent activation of both SAMK and HAMK is mimicked at 25°C by destabilizers of microfilaments and microtubules, latrunculin B and oryzalin, respectively; but is blocked by jasplakinolide, a stabilizer of actin microfilaments. Activation of SAMK or HAMK by temperature, chemically modulated membrane fluidity, or by cytoskeleton destabilizers is inhibited by blocking the influx of extracellular calcium. Activation of SAMK or HAMK is also prevented by an antagonist of calcium‐dependent protein kinases (CDPKs). In summary, our data indicate that cold and heat are sensed by structural changes in the plasma membrane that translates the signal via cytoskeleton, Ca2+ fluxes and CDPKs into the activation of distinct MAPK cascades.
Environmental and auxin regulation of wood formation involves members of the Aux/IAA gene family in hybrid aspen
Authors: Moyle Richard , Schrader Jarmo , Stenberg Anneli , Olsson Olof , Saxena Sangeeta , Sandberg Göran , Bhalerao Rishikesh P
Published: March 2003

Indole acetic acid (IAA/auxin) profoundly affects wood formation but the molecular mechanism of auxin action in this process remains poorly understood. We have cloned cDNAs for eight members of the Aux/IAA gene family from hybrid aspen (Populus tremula L. × Populus tremuloides Michx.) that encode potential mediators of the auxin signal transduction pathway. These genes designated as PttIAA1‐PttIAA8 are auxin inducible but differ in their requirement of de novo protein synthesis for auxin induction. The auxin induction of the PttIAA genes is also developmentally controlled as evidenced by the loss of their auxin inducibility during leaf maturation. The PttIAA genes are differentially expressed in the cell types of a developmental gradient comprising the wood‐forming tissues. Interestingly, the expression of the PttIAA genes is downregulated during transition of the active cambium into dormancy, a process in which meristematic cells of the cambium lose their sensitivity to auxin. Auxin‐regulated developmental reprogramming of wood formation during the induction of tension wood is accompanied by changes in the expression of PttIAA genes. The distinct tissue‐specific expression patterns of the auxin inducible PttIAA genes in the cambial region together with the change in expression during dormancy transition and tension wood formation suggest a role for these genes in mediating cambial responses to auxin and xylem development.
Complex transgene locus structures implicate multiple mechanisms for plant transgene rearrangement
Authors: Svitashev Sergei K. , Pawlowski Wojciech P. , Makarevitch Irina , Plank David W. , Somers David A.
Published: March 2003

To more fully characterize the internal structure of transgene loci and to gain further understanding of mechanisms of transgene locus formation, we sequenced more than 160 kb of complex transgene loci in two unrelated transgenic oat (Avena sativa L.) lines transformed using microprojectile bombardment. The transgene locus sequences from both lines exhibited extreme scrambling of non‐contiguous transgene and genomic fragments recombined via illegitimate recombination. A perfect direct repeat of the delivered DNA, and inverted and imperfect direct repeats were detected in the same transgene locus indicating that homologous recombination and synthesis‐dependent mechanism(s), respectively, were also involved in transgene locus rearrangement. The most unexpected result was the small size of the fragments of delivered and genomic DNA incorporated into the transgene loci via illegitimate recombination; 50 of the 82 delivered DNA fragments were shorter than 200 bp. Eleven transgene and genomic fragments were shorter than the DNA lengths required for Ku‐mediated non‐homologous end joining. Detection of these small fragments provided evidence that illegitimate recombination was most likely mediated by a synthesis‐dependent strand‐annealing mechanism that resulted in transgene scrambling. Taken together, these results indicate that transgene locus formation involves the concerted action of several DNA break‐repair mechanisms.
Ozone‐induced ethylene production is dependent on salicylic acid, and both salicylic acid and ethylene act in concert to regulate ozone‐induced cell death
Authors: Rao Mulpuri V. , Lee Hyung‐il , Davis Keith R.
Published: March 2003

Ethylene is known to influence plant defense responses including cell death in response to both biotic and abiotic stress factors. However, whether ethylene acts alone or in conjunction with other signaling pathways is not clearly understood. Ethylene overproducer mutants, eto1 and eto3, produced high levels of ethylene and developed necrotic lesions in response to an acute O3 exposure that does not induce lesions in O3‐tolerant wild‐type Col‐0 plants. Treatment of plants with ethylene inhibitors completely blocked O3‐induced ethylene production and partially attenuated O3‐induced cell death. Analyses of the responses of molecular markers of specific signaling pathways indicated a relationship between salicylic acid (SA)‐ and ethylene‐signaling pathways and O3 sensitivity. Both eto1 and eto3 plants constitutively accumulated threefold higher levels of total SA and exhibited a rapid increase in free SA and ethylene levels prior to lesion formation in response to O3 exposure. SA pre‐treatments increased O3 sensitivity of Col‐0, suggesting that constitutive high SA levels prime leaf tissue to exhibit increased magnitude of O3‐induced cell death. NahG and npr1 plants compromised in SA signaling failed to produce ethylene in response to O3 and other stress factors suggesting that SA is required for stress‐induced ethylene production. Furthermore, NahG expression in the dominant eto3 mutant attenuated ethylene‐dependent PR4 expression and rescued the O3‐induced HR (hypersensitive response) cell death phenotype exhibited by eto3 plants. Our results suggest that both SA and ethylene act in concert to influence cell death in O3‐sensitive genotypes, and that O3‐induced ethylene production is dependent on SA.
Both antisense and sense expression of biotin carboxyl carrier protein isoform 2 inactivates the plastid acetyl‐coenzyme A carboxylase in Arabidopsis thaliana
Authors: Thelen Jay J. , Ohlrogge John B.
Published: March 2003

To further characterize the role of biotin carboxyl carrier protein isoform 2 (BCCP2) in acetyl‐coenzyme A carboxylase (ACCase) function and fatty acid biosynthesis, plants with reduced or increased expression of this protein were characterized. Analysis of 38 independent Arabidopsis lines expressing antisense BCCP2 transcript behind a constitutive promoter showed no significant phenotype, though antisense transcript was highly expressed. In developing seed, BCCP2 protein was reduced by an average of 38% resulting in a 9% average decrease in fatty acid content in mature seed. Over‐expression of BCCP2 behind a seed‐specific napin promoter increased the amount of holo‐BCCP2 in developing seed by an average of twofold, as determined with anti‐biotin antibodies. Surprisingly, the average fatty acid content of T2 seed from over‐expression lines was 23% lower than wild‐type seed. These plants also exhibited reduced seed setting in 18 of 20 T1 lines which was coincident with increased individual seed mass. Quantification of total BCCP2 in developing siliques using anti‐BCCP2 antibodies indicated that as much as 60% of total plastidial BCCP2 was in the non‐biotinylated form (apo‐BCCP2). Although apo‐BCCP2 was highly over‐expressed in developing seed, accumulation of other ACCase subunits was unaffected. The specific activity of ACCase was up to 65% lower in developing seed of over‐expression lines versus wild type. This was attributed to the assembly of biologically inactive (non‐biotinylated) ACCase complexes. Consistent with ACCase exerting control over de novo fatty acid synthesis, reduced activity in developing seed resulted in lower oil content, altered fatty acid composition and reduced seed setting.
The impact of oxidative stress on Arabidopsis mitochondria
Authors: Sweetlove L.J. , Heazlewood J.L. , Herald V. , Holtzapffel R. , Day D.A. , Leaver C.J. , Millar A.H.
Published: March 2003

Treatment of Arabidopsis cell culture for 16 h with H2O2, menadione or antimycin A induced an oxidative stress decreasing growth rate and increasing DCF fluorescence and lipid peroxidation products. Treated cells remained viable and maintained significant respiratory rates. Mitochondrial integrity was maintained, but accumulation of alternative oxidase and decreased abundance of lipoic acid‐containing components during several of the treatments indicated oxidative stress. Analysis of the treatments was undertaken by IEF/SDS‐PAGE, comparison of protein spot abundances and tandem mass spectrometry. A set of 25 protein spots increased >3‐fold in H2O2/menadione treatments, a subset of these increased in antimycin A‐treated samples. A set of 10 protein spots decreased significantly during stress treatments. A specific set of mitochondrial proteins were degraded by stress treatments. These damaged components included subunits of ATP synthase, complex I, succinyl CoA ligase, aconitase, and pyruvate and 2‐oxoglutarate dehydrogenase complexes. Nine increased proteins represented products of different genes not found in control mitochondria. One is directly involved in antioxidant defense, a mitochondrial thioredoxin‐dependent peroxidase, while another, a thioredoxin reductase‐dependent protein disulphide isomerase, is required for protein disulfide redox homeostasis. Several others are generally considered to be extramitochondrial but are clearly present in a highly purified mitochondrial fraction used in this study and are known to play roles in stress response. Using H2O2 as a model stress, further work revealed that this treatment induced a protease activity in isolated mitochondria, putatively responsible for the degradation of oxidatively damaged mitochondrial proteins and that O2 consumption by mitochondria was significantly decreased by H2O2 treatment.
Molecular analysis of the stylar‐expressed Solanum chacoense small asparagine‐rich protein family related to the HT modifier of gametophytic self‐incompatibility in Nicotiana
Authors: O'Brien M. , Kapfer C. , Major G. , Laurin M. , Bertrand C. , Kondo K. , Kowyama Y. , Matton D. P.
Published: March 2003

Gametophytic self‐incompatibility (GSI) systems involving the expression of stylar ribonucleases have been described and extensively studied in many plant families including the Solanaceae, Rosaceae and Scrophulariaceae. Pollen recognition and rejection is governed in the style by specific ribonucleases called S‐RNases, but in many self‐incompatibility (SI) systems, modifier loci that can modulate the SI response have been described at the genetic level. Here, we present at the molecular level, the isolation and characterization of two Solanum chacoense homologues of the Nicotiana HT modifier that had been previously shown to be necessary for the SI reaction to occur in N. alata (McClure et al., 1999). HT homologues from other solanaceous species have also been isolated and a phylogenetic analysis reveals that the HT genes fall into two groups. In S. chacoense, these small proteins named ScHT‐A and ScHT‐B are expressed in the style and are developmentally regulated during anthesis identically to the S‐RNases as well as following compatible and incompatible pollination. To elucidate the precise role of each HT isoform, antisense ScHT‐A and RNAi ScHT‐B lines were generated. Conversion from SI to self‐compatibility (SC) was only observed in RNAi ScHT‐B lines with reduced levels of ScHT‐B mRNA. These results confirm the role of the HT modifier in solanaceous SI and indicate that only the HT‐B isoform is directly involved in SI.
A natural recessive resistance gene against potato virus Y in pepper corresponds to the eukaryotic initiation factor 4E (eIF4E)
Authors: Ruffel Sandrine , Dussault Marie‐Hélène , Palloix Alain , Moury Benoît , Bendahmane Abdelhafid , Robaglia Christophe , Caranta Carole
Published: March 2003

We show here that the pvr2 locus in pepper, conferring recessive resistance against strains of potato virus Y (PVY), corresponds to a eukaryotic initiation factor 4E (eIF4E) gene. RFLP analysis on the PVY‐susceptible and resistant pepper cultivars, using an eIF4E cDNA from tobacco as probe, revealed perfect map co‐segregation between a polymorphism in the eIF4E gene and the pvr2 alleles, pvr21 (resistant to PVY‐0) and pvr22 (resistant to PVY‐0 and 1). The cloned pepper eIF4E cDNA encoded a 228 amino acid polypeptide with 70–86% nucleotide sequence identity with other plant eIF4Es. The sequences of eIF4E protein from two PVY‐susceptible cultivars were identical and differed from the eIF4E sequences of the two PVY‐resistant cultivars Yolo Y (YY) (pvr21) and FloridaVR2 (F) (pvr22) at two amino acids, a mutation common to both resistant genotypes and a second mutation specific to each. Complementation experiments were used to show that the eIF4E gene corresponds to pvr2. Thus, potato virus X‐mediated transient expression of eIF4E from susceptible cultivar Yolo Wonder (YW) in the resistant genotype YY resulted in loss of resistance to subsequent PVY‐0 inoculation and transient expression of eIF4E from YY (resistant to PVY‐0; susceptible to PVY‐1) rendered genotype F susceptible to PVY‐1. Several lines of evidence indicate that interaction between the potyvirus genome‐linked protein (VPg) and eIF4E are important for virus infectivity, suggesting that the recessive resistance could be due to incompatibility between the VPg and eIF4E in the resistant genotype.
The circadian clock regulated RNA‐binding protein AtGRP7 autoregulates its expression by influencing alternative splicing of its own pre‐mRNA
Authors: Staiger Dorothee , Zecca Laura , Kirk Dominika A. Wieczorek , Apel Klaus , Eckstein Luca
Published: March 2003

The clock‐regulated RNA‐binding protein AtGRP7 is part of a negative feedback circuit through which the protein influences circadian oscillations of its own transcript. Constitutive overexpression of AtGRP7 in transgenic plants leads to the appearance of a low amount of an alternatively spliced Atgrp7 transcript with a premature stop codon. It is generated by the use of a 5′ cryptic splice site in the middle of the intron at the expense of the fully spliced mRNA, indicating a role for AtGRP7 in splice site selection. Accelerated decay of this transcript accounts for its low steady state abundance. This implicates a mechanism for the AtGRP7 feedback loop: Atgrp7 expression is downregulated, as AtGRP7 protein accumulates over the circadian cycle, partly by the generation of an alternate transcript that due to its instability does not accumulate to high levels and does not produce a functional protein. Recombinant AtGRP7 protein specifically interacts with the 3′ untranslated region and the intron of its transcript, suggesting that the shift in splice site selection and downregulation involves binding of AtGRP7 to its pre‐mRNA. AtGRP7 also influences the choice of splice sites in the Atgrp8 transcript encoding a related RNA‐binding protein, favoring the production of an alternatively spliced, unstable Atgrp8 transcript. This conservation points to the importance of this regulatory mechanism to control the level of the clock‐regulated glycine‐rich RNA‐binding proteins and shows how AtGRP7 can control abundance of target transcripts.
Molecular properties of the putative nitrogen sensor PII from Arabidopsis thaliana
Authors: Smith Catherine S. , Weljie Aalim M. , Moorhead Greg B. G.
Published: March 2003

Although the signal sensing protein PII is well known to play a central role in bacterial nitrogen metabolism, the structure and function of PII in plants remains only partially understood. Comparative modeling was undertaken based on the high degree of amino acid identity between Escherichia coli and Arabidopsis PII. The mature Arabidopsis PII predicted structure superimposes very well onto the E. coli PII structure (Cα root mean square deviation < 0.4 Å). The model of the highly conserved T‐loop suggests a molecular mechanism by which the plant PII may regulate putative post‐translational modification in response to metabolite binding. Consistent with the presence of key conserved residues necessary for trimer formation, gel filtration showed the oligomeric structure of Arabidopsis thaliana PII to be a homotrimer. We have demonstrated that Arabidopsis PII binds to the small molecules, ATP, ADP, 2KG, and with lesser affinity to OAA, using isothermal titration calorimetry. We have determined the metabolite dissociation constants and compared these with known physiological concentrations of these metabolites in the plant to identify the Arabidopsis PII effector molecules and their possible roles. We predict that the plant PII is likely continually bound by ATP, and its ligand‐bound state only varying with respect to the degree of 2KG binding. Based on our in vitro binding studies, the function of plant PII as a 2KG sensor is suggested.
The OsTB1 gene negatively regulates lateral branching in rice
Authors: Takeda Taito , Suwa Yuko , Suzuki Makoto , Kitano Hidemi , Ueguchi‐Tanaka Miyako , Ashikari Motoyuki , Matsuoka Makoto , Ueguchi Chiharu
Published: March 2003

Although the shoot apical meristem (SAM) is ultimately responsible for post‐embryonic development in higher plants, lateral meristems also play an important role in determining the final morphology of the above‐ground part. Axillary buds developing at the axils of leaves produce additional shoot systems, lateral branches. The rice TB1 gene (OsTB1) was first identified based on its sequence similarity with maize TEOSINTE BRANCHED 1 (TB1), which is involved in lateral branching in maize. Both genes encode putative transcription factors carrying a basic helix–loop–helix type of DNA‐binding motif, named the TCP domain. The genetic locus of OsTB1 suggested that OsTB1 is a real counterpart of maize TB1. Transgenic rice plants overexpressing OsTB1 exhibited markedly reduced lateral branching without the propagation of axillary buds being affected. We also demonstrated that a rice strain carrying a classical morphological marker mutation, fine culm 1 (fc1), contain the loss‐of‐function mutation of OsTB1 and exhibits enhanced lateral branching. Expression of OsTB1, as examined with a putative promoter–glucuronidase (GUS) gene fusion, was observed throughout the axillary bud, as well as the basal part of the shoot apical meristem, vascular tissues in the pith and the lamina joint. Taking these data together, we concluded that OsTB1 functions as a negative regulator for lateral branching in rice, presumably through expression in axillary buds.
Cell cycle‐dependent association ofArabidopsisactin‐related proteins AtARP4 and AtARP7 with the nucleus
Authors: Kandasamy Muthugapatti K. , McKinney Elizabeth C. , Meagher Richard B.
Published: March 2003

Arabidopsisencodes at least eight actin‐related proteins (ARPs) most of which have orthologs in other distant organisms. To gain insight into the role of ARPs in plants, we have examined the spatial expression and subcellular distribution of two highly divergentArabidopsisARPs, AtARP4 and AtARP7. AtARP4 is a homolog of human BAF53 and yeast Arp4, and AtARP7 is a novel, ancient and plant‐specific actin‐related protein that is not distinctly related to any known ARPs from other kingdoms. Analysis of both these proteins with AtARP4‐ and AtARP7‐specific antibodies revealed that they were most abundant in young meristematic and floral tissues, but were expressed constitutively in all organs and cell types irrespective of their developmental stage. Immunofluorescence studies showed that both AtARP4 and AtARP7 were localized predominantly to the nucleus during interphase. In mitotic cells lacking a nuclear envelope (e.g. metaphase, anaphase, and early telophase stages), these ARPs were excluded from the condensed chromosomes and dispersed throughout the cytoplasm. In contrast, a putativeArabidopsishistone H2B protein remained associated with the interphase nuclei as well as chromosomes throughout the cell cycle. Based on our results and data on the yeast ortholog of AtARP4, these two nuclear plant ARPs may be involved in the modulation of chromatin structure and transcriptional regulation mainly in interphase cells.
Transfusing platelets 2 h after the completion of amphotericin‐B decreases its detrimental effect on transfused platelet recovery and survival
Authors: Hussein  , Fletcher  , Long  , Zuccaro  , Bolwell  , Hoeltge
Published: March 2003

Platelet transfusion support is required during bone marrow aplasia following ablative chemotherapy and bone marrow progenitor cell transplantation (BMT). Amphotericin‐B is frequently given to these patients, both therapeutically and prophylactically, and has been described to have a negative impact on the results of platelet transfusions. We conducted a prospective study of the effect of amphotericin‐B on transfused platelet recovery and survival in 81 BMT or acute leukaemia patients. One hundred and ninety‐five platelet transfusions administered to 81 consecutive patients were analysed. The platelets were transfused 2 h after the completion of amphotericin‐B. Using this schedule resulted in no effect of amphotericin‐B on platelet recovery or survival, although platelet increments were modestly depressed in patients receiving high‐ vs. low‐dose amphotericin‐B. We conclude that the timing of amphotericin‐B infusion be evaluated in patients demonstrating poor platelet recovery and survival. Transfusing platelets at least 2 h after the completion of amphotericin‐B decreases the detrimental effect of this antifungal agent on transfused platelet recovery and survival.
Report on the 1997 International Society of Blood Transfusion Workshop for Genotyping of Platelet Alloantigens
Authors: Panzer  , Kaplan  ,   on behalf of the Platelet and Granulocyte Workshop ISBT
Published: March 2003

The results from the 1997 International human platelet antigen‐ (HPA‐) genotyping workshop are reported. Seventeen institutions participated to genotype for HPA polymorphisms of seven DNA samples. Most investigators used the polymerase chain reaction (PCR) — restriction fragment length polymorphism (RFLP) and PCR‐sequence specific priming (SSP). A few participants applied both methods to confirm their results. All participants reported results for HPA‐1, ‐2, ‐3 and ‐5 allotyping, while HPA‐4 was determined in 12 and HPA‐6 in nine laboratories, respectively. For HPA‐1 to ‐6 there were 19 deviations from the consensus result. These deviations were not associated with any particular technique or allotype. Most frequently, however, deviation were reported for HPA‐5 typing. These results were discussed at a meeting at the Joint Congress of the ISBT and the Deutsche Gesellschaft für Transfusionsmedizin und Immunhämatologie in Frankfurt, Germany, in September 1997. Based on the results and the comments, further workshops for HPA‐allotyping are mandatory for quality assurance.
Guidelines for irradiation of blood and blood components to prevent post‐transfusion graft‐vs.‐host disease in Japan
Authors: Asai T. , Inaba S. , Ohto H. , Osada K. , Suzuki G. , Takahashi K. , Tadokoro K. , Minami M.
Published: March 2003

. The Japanese Red Cross analysed the results of questionnaires sent in 1993 regarding post‐transfusion graft‐vs.‐host disease (PT‐GVHD) from hospitals; the majority of patients with PT‐GVHD in 1993 were transfused for cardiovascular or cancer surgery, and about 10 patients had died yearly from PT‐GVHD in the following few years.

The Japan Society of Blood Transfusion (JSBT) organized a subcommittee for the prevention of PT‐GVHD, and issued a fourth version of guidelines for the irradiation of blood to prevent PT‐GVHD. These guidelines recommended transfusion of irradiated blood for cardiosurgery, cancer surgery, elderly recipients and severe trauma, as well as congenital immunodeficient recipients, newborn infants and other immunocompromised patients. Also recommended was irradiation not only of blood within 72 h after collection but also of blood stored for 14 days. Reported PT‐GVHD has diminished to a few cases in recent years.
Characterization of a novel hepatitis B virus mutant: demonstration of mutation‐induced hepatitis B virus surface antigen group specific ‘a’ determinant conformation change and its application in diagnostic assays
Authors: Kfoury Baz E. M. , Zheng J. , Mazuruk K. , Van Le A. , Peterson D. L.
Published: March 2003

. We report and characterize a novel mutant of the hepatitis B virus surface antigen (HBsAg). The mutant was isolated from a symptomatic patient who was found to be persistently positive for both HBsAg and anti‐hepatitis B surface antibody (anti‐HBs) and a long‐lasting anti‐HBc (core) IgM. Due to the unusual immune serological profile, polymerase chain reaction and sequencing were performed and revealed a genotype D mutant HBV (LBN). Aligned with known HBsAg sequences from GenBank, the LBN variant matched to consensus subtype ayw2 and revealed five mutation positions. Interestingly, none of the mutations was found within the group‐specific ‘a’ determinant region (124–147) and, specifically, two of the five mutations, T118K and P120Q, were located only a few amino acids adjacent to the 124–147 region. Using a panel of six monoclonal antibodies and vaccine raised human neutralizing antibodies, the recombinant wild‐type HBsAg and the novel variant LBN HBsAg were investigated for their immunological reactivity. Vaccine raised human anti‐HBs showed less reactivity to the variant LBN HBsAg than to wild‐type HBsAg in enzyme immune assays. Furthermore, our observations demonstrate the influence of adjacent residues on the group‐specific ‘a’ determinant structural configuration; this resulted in severe antigenic changes of the immunodominant region as well as in the subtype serology. The importance of the variant LBN lies in the observation that mutations close to the ‘a’ determinant can change the immunodominant region structure and therefore alter the group specific determinant antigenicity even though no mutations are present within this region. Hence, the classical definition of the ‘a’ determinant cluster may need to be extended to cover a broader region to include the requirement of adjacent amino acids to support its conformation. In conclusion, by understanding the HBsAg major immunodominant region structure and by using a combination of antibodies with specificity covering all key mutation locations, maximal anti‐HBs‐based protection, and highly sensitive diagnostic assays can be achieved.
Platelet transfusion in the management of severe thrombocytopenia in neonatal intensive care unit patients
Authors: Murray N. A. , Howarth L. J. , McCloy M. P. , Letsky E. A. , Roberts I. A. G.
Published: March 2003

Summary Platelet transfusions are frequently given to neonatal intensive care unit (NICU) patients with severe thrombocytopenia (platelets less than 50 × 109 L−1) but no study has assessed whether this is clinically appropriate. To address this we conducted a retrospective review of platelet transfusion practice in patients developing severe thrombocytopenia over 3 years in a single NICU. Out of 901 admissions, 53 (6%) developed severe thrombocytopenia. Twenty‐seven neonates received a total of 63 platelet transfusions, the main triggers being: platelet count less than 30 × 109 L−1 (all patients), or less than 50 × 109 L−1 in those with previous haemorrhage or clinical instability. No major haemorrhage occurred during severe thrombocytopenia either in neonates in whom platelet transfusions were withheld (26/53) or in neonates given platelets who survived to discharge (22/27). Five preterm neonates given platelets died but all had overwhelming sepsis or necrotizing enterocolitis and none died directly as a result of haemorrhage.

Although the widely used liberal triggers for neonatal platelet transfusion highlighted in this review reflect available guidelines, and represent cautious (‘safe’) haemostatic practice, they are likely to result in unnecessary transfusion for a significant number of NICU patients. Improved practice requires definition of a safe lower limit for platelet count in stable neonates; effective platelet transfusion strategies for sick neonates; and improved therapies for conditions precipitating severe thrombocytopenia.
In vitroantiviral and antibacterial activity of commercial intravenous immunoglobulin preparations – a potential role for adjuvant intravenous immunoglobulin therapy in infectious diseases*
Authors: Krause I. , Wu R. , Sherer Y. , Patanik M. , Peter J. B. , Shoenfeld Y.
Published: March 2003

SummaryThe identification of specific antimicrobial activity of intravenous immunoglobulin (IVIG) preparations against particular microbial pathogens can assist in determining their therapeutic potential for specific infectious diseases. We analysed five different commercial IVIG preparations for the presence of antibodies directed against a large panel of viral, bacterial, fungal and parasitic pathogens. All IVIG batches contained high activity against herpesviruses types 1, 2, 6 and 7, as well as against varicella zoster virus, Epstein‐Barr virus (EBV), measles, mumps, rubella and parvovirus B19. Some IVIG batches also had a significant activity against adenovirus and Saint Louis encephalitis virus. The IVIGs held high activity against several bacterial pathogens, includingMycoplasma pneumonia,Chlamydia pneumonia,Helicobacter pyloriand tetanus. No activity was found against various parasitic and fungal pathogens. Our findings may provide further support for the use of IVIG for the prevention and treatment of infections caused by specific viral and bacterial pathogens.
Limited efficacy of universal leucodepletion in reducing the incidence of febrile nonhaemolytic reactions in red cell transfusions
Authors: Ibojie J. , Greiss M. A. , Urbaniak S. J.
Published: March 2003

Summary  This article demonstrates a 62% reduction in the number of febrile nonhaemolytic transfusion reactions (FNHTRs) and 50% reduction in febrile reaction rate associated with red cell transfusions following graded introduction of universal leucodepletion. Though this is a statistically significant reduction (P = 0·009), it shows limited efficacy in abrogating this complication. We also found a reduction in the proportion of cases of FNHTRs with lymphocytotoxic antibodies over the period studied from 54% in 1998, 28% in 1999 to 23% in 2000. This corresponds to a relative increase in the number of febrile reactions without human leucocyte antigen (HLA) antibodies following full implementation of universal leucodepletion, as the total number of reported reactions actually fell considerably during the period. The increase in the number of cases without HLA antibodies was directly proportional to the increase in the number of leucodepleted units used.
Structural characterization of the epitope recognized by the new anti‐Fy6 monoclonal antibody NaM185‐2C3
Authors: Wasniowska K. , Petit‐LeRoux Y. , Tournamille C. , Le van Kim C. , Cartron J. P. , Colin Y. , Lisowska E. , Blanchard D.
Published: March 2003

Summary  The epitope recognized by a new anti‐Fy6 monoclonal antibody (MoAb) (clone name: NaM185‐2C3) was characterized using peptides synthesized on pins (Epitope scanning kit). The clone was obtained from splenocytes of mice immunized with CHO cells expressing the recombinant Duffy glycoprotein. NaM185‐2C3 recognized a linear epitope, the essential portion of which was pentapeptide Phe‐Glu‐Asp‐Val‐Trp comprising amino acid residues 22–26 of the main (336 aa) isoform of the Duffy antigen. All the amino acid residues of the epitope, except Asp, were essential for the antibody‐binding, because they could not be replaced by any or most other amino acid residues. The Asp residue could be replaced by most other amino acid residues and its replacement by some amino acid residues gave a distinct increase in the antibody‐binding. The MoAb NaM185–2C3, similarly as other anti‐Fy6 antibodies, inhibits interleukin (IL)‐8‐binding to the Duffy antigen. A part of the results was presented at ISBT meeting (Blanchard et al., 1998, Vox Sanguinis, 74, S1, Abstract no. 71).
Nucleic acid testing for emerging viral infections
Authors: Allain J.‐P. , Thomas I. , Sauleda S.
Published: March 2003

Summary. The development of new technologies leads to the discovery of new viruses. For each of these new infectious agents, relevance to transfusion, including transmissibility by transfusion, pathogenicity, prevalence in blood donors, persistence and the availability of screening assays needs to be assessed. Since 1995, one virus and a new family of viruses have been identified. GB virus‐C/hepatitis G virus (GBV‐C/HGV), a flavi virus with some homology with and epidemiological features of HCV, is not related to post‐transfusion hepatitis but seems to positively interfere with human immunodeficiency virus replication. Human circoviruses include TT virus (TTV) and SEN‐V. Both are highly variable, constituting a large family of distantly related viruses. They appear ubiquitous, infecting humans very early in life and are largely persistent. No clinical symptoms or pathogenicity is associated with TTV, but SEN‐V might be associated with some non‐A–E post‐transfusion hepatitis. Parvovirus B19 has been known for many years, but its transmission to recipients of plasma derivatives despite viral inactivation raised the issue of screening plasma pools by nucleic acid testing. Most fractionators quantify B19 DNA in plasma pools to ensure a viral load of <104 IU mL−1.
Neuropsychiatric manifestations after mefloquine therapy for Plasmodium falciparum malaria: comparing a retrospective and a prospective study
Authors: Rønn A. M. , Rønne‐Rasmussen J. , Gøtzsche P. C. , Bygbjerg I. C.
Published: March 2003

Mefloquine has been increasingly used for treatment of chloroquine‐resistant malaria since its introduction in the late 1970s. In 1987 the first case of toxic encephalopathy was published, and in 1989 the WHO initiated reporting and investigation of neuropsychiatric adverse reactions of mefloquine. Neuropsychiatric adverse drug reactions are now well documented. We compared an open prospective 3 year study including all patients with P. falciparum treated with mefloquine with an earlier published, retrospective study on a comparable population from our department covering the period up to 1989.dollar;>In the retrospective study neuropsychiatric adverse effects were not specifically asked for, while in the prospective study possible adverse reactions were registered daily according to a specified questionnaire. No case of neuropsychiatric adverse reaction was registered in the retrospective study. In the prospective study, 28% had one or more neuropsychiatric adverse reactions, although severity was mostly mild to moderate. Other adverse reactions occurred in 96% in the retrospective study compared to 81% in the prospective study. In conclusion: one often finds only what one looks for, e.g adverse events may be overlooked for a decade, if relatively uncommon. This report also shows that retro‐ and prospective studies may give very different results.
Allergic bronchopulmonary aspergillosis: disease pattern in central Arabia
Authors: Mobeireek Abdullah , El‐Rab Mohamed O. Gad , Joharjy Ismail , Al‐Sohaibani Mohammed , Ashour Mahmood
Published: March 2003

Despite a high prevalence of asthma in Saudi Arabia, allergic bronchopulmonary aspergillosis (ABPA) has not been reported. We reviewed the medical records in a large university hospital in Saudi Arabia where thousands of asthmatics are being followed up. Over a 9‐year period starting January 1986, the diagnosis of ABPA was made in 10 patients only. Delay in diagnosis was common and in some patients the disease was confused with fungal pneumonia, tuberculosis or tumours. Aspergillus fumigatus was isolated from one patient only and different Aspergillus species were cultured from respiratory secretions of the others. Corticosteroids were uniformly effective in all patients with active disease. Low humidity may account for this apparent rarity of ABPA, although it is possible that some cases are overlooked. Further work is needed on the prevalent fungi in the Arabian environment and their potential health effects and particularly on the prevalence of allergic bronchopulmonary fungal disease.
Seasonal density, sporozoite rates and entomological inoculation rates of Anopheles gambiae and Anopheles funestus in a high‐altitude sugarcane growing zone in western kenya
Authors: Shililu  , Maier  , Seitz  , Orago
Published: March 2003

Summary An entomological study was conducted on vectors of malaria and their relative contribution to Plasmodium falciparum transmission in Mumias, a high‐altitude site and large‐scale sugarcane growing zone in Kakamega district, western Kenya. Anopheles gambiae s.l., the predominant vector species, represented 84% (n= 2667) of the total Anopheles mosquitoes collected with An. funestus comprising only 16%. Polymerase chain reaction (PCR) identified all 600 specimens of the An. gambiae complex tested as An. gambiaesensu stricto, an indication that it is the only sibling species represented in the high‐altitude sites in western Kenya. Plasmodium falciparum sporozoite rates of 6.3% (133/2118) for An. gambiae s.l. and 9.5% (38/402) for An. funestus by ELISA were obtained in Mumias. None of 1600 mosquitoes tested for P. malariae sporozoites was positive. ELISA tests of mosquito blood meals indicated a high tendency of anthropophagy, a behaviour contributing significantly to malaria transmission by the vector species, with 95.9%, 4.86% and 0.2% having taken at least one blood meal on human, bovine and avian hosts, respectively.  Malaria transmission intensity was low as revealed by the low entomological inoculation rates (EIR) recorded. The EIR values for An. gambiae s.l. were 29.2 infective bites per person per year (ib/p/year) and 17.5 ib/p/year for An. funestus in Mumias. The highest inoculation rate for both vector species was 7.0 ib/p/month in July. Plasmodium falciparum parasite rate among asymptomatic children was 55.4% and 44% in the wet (July–September) and dry (December–February) seasons, respectively. These results indicate that malaria transmission intensity in the high‐altitude site is low but perennial, with transmission being maintained by An. gambiae s.s. and An. funestus.
Predicting key malaria transmission factors, biting and entomological inoculation rates, using modelled soil moisture in Kenya
Authors: Patz Jonathan A.
Published: March 2003

While malaria transmission varies seasonally, large inter‐annual heterogeneity of malaria incidence occurs. Variability in entomological parameters, biting rates and entomological inoculation rates (EIR) have been strongly associated with attack rates in children. The goal of this study was to assess the weather's impact on weekly biting and EIR in the endemic area of Kisian, Kenya. Entomological data collected by the U.S. Army from March 1986 through June 1988 at Kisian, Kenya was analysed with concurrent weather data from nearby Kisumu airport. A soil moisture model of surface‐water availability was used to combine multiple weather parameters with landcover and soil features to improve disease prediction. Modelling soil moisture substantially improved prediction of biting rates compared to rainfall; soil moisture lagged two weeks explained up to 45% of An. gambiae biting variability, compared to 8% for raw precipitation. For An. funestus, soil moisture explained 32% variability, peaking after a 4‐week lag. The interspecies difference in response to soil moisture was significant (P < 0.00001). A satellite normalized differential vegetation index (NDVI) of the study site yielded a similar correlation (r2= 0.42 An. gambiae). Modelled soil moisture accounted for up to 56% variability of An. gambiae EIR, peaking at a lag of six weeks. The relationship between temperature and An. gambiae biting rates was less robust; maximum temperature r2=−0.20, and minimum temperature r2= 0.12 after lagging one week. Benefits of hydrological modelling are compared to raw weather parameters and to satellite NDVI. These findings can improve both current malaria risk assessments and those based on El Niño forecasts or global climate change model projections.
Review of varicella zoster seroepidemiology in India and South‐east Asia
Authors: Lee
Published: March 2003

Summary Varicella zoster virus (VZV) predominantly affects children in temperate countries, with near‐universal seroconversion occurring by late childhood. However, in tropical regions, VZV infection is common in adolescents and adults. This review identifies age‐related VZV seroprevalence patterns in a number of Asian countries which indicate that seroconversion in tropical countries occurs at a later age than in temperate countries. Seasonal and regional variations in acute disease within some Asian countries suggest that temperate climates might favour transmission of the varicella virus, with incidence peaking during cooler months and in cooler, more temperate regions. VZV infection is often more severe in adults than in children, suggesting that tropical countries may be at risk of greater morbidity and mortality as a result of later‐age seroconversion. Susceptibility of pregnant women and their infants, and of people infected with HIV/AIDS is also cause for concern. Vaccination may be beneficial in reducing the impact of VZV in Asian populations.
Applying medical anthropology in the control of infectious disease
Authors: Manderson Lenore
Published: March 2003

This paper focuses on two roles of anthropology in the control of infectious disease. The first is in identifying and describing concerns and understandings of disease, including local knowledge of cause and treatment relevant to disease control. The second is in translating these local concerns into appropriate health interventions, for example, by providing information to be incorporated in education and communication strategies for disease control. Problems arise in control programmes with competing knowledge and value systems. Anthropology's role conventionally has been in the translation of local concepts of illness and treatment, and the adaptation of biomedical knowledge to fit local aetiologies. Medical anthropology plays an important role in examining the local context of disease diagnosis, treatment and prevention, and the structural as well as conceptual barriers to improved health status. National (and international) public health goals which respect local priorities are uncommon, and generic health goals rarely coincide with specific country and community needs. The success of interventions and control programmes is moderated by local priorities and conditions, and sustainable interventions need to acknowledge and address country‐specific social, economic and political circumstances.
Methods and meanings in anthropological, epidemiological and clinical encounters: the case of sexually transmitted disease and human immunodeficiency virus control and prevention in India
Authors: Lambert Helen
Published: March 2003

Rapid assessment procedures informed by anthropology are increasingly commonly being utilised in the formulation of a range of international disease control and prevention strategies. Refinement of these rapid procedures has narrowly focused on their methods, in contrast with the broader potential scope of context‐based anthropological contributions. The control of HIV and other sexually transmitted diseases (STDs) in India is used to illustrate the potential role of anthropologically informed insights and investigations in the development of effective interventions, with particular reference to the examples of syndromic management, symptom presentation and treatment provision. The paper argues for anthropological epidemiological and clinical studies to be properly integrated in order to fulfil their substantial potential for interdisciplinary illumination.
Syndromic management of sexually transmitted diseases: is it rational or scientific?
Authors: Bosu William K.
Published: March 2003

Summary Control of sexually transmitted diseases improves the reproductive and sexual health of afflicted individuals and lowers the community burden of HIV. Preventing STDs or their complications requires health education, condom promotion and effective case management. Clinical diagnosis is frequently incorrect while laboratory‐confirmed aetiological diagnosis is expensive. In resource‐poor countries which lack trained personnel and laboratory facilities, syndromic management remains a rational approach to STD care. Although the approach is simplistic in design, its successful implementation requires regular monitoring and evaluation of protocols as well as supervision and training of clinicians.