bluuebunny/crossref_metadata_2025_split huggingface dataset:
Agoraphobia: nurse therapist‐facilitated self‐help manual
Authors: Lovell Karina , Cox Debbie , Garvey Rachel , Raines David , Richards David , Conroy Patrick , Repper Dean 
Published: August 2003

Background. Agoraphobia is a common and disabling mental health disorder. Substantial evidence supports the use of cognitive behaviour therapy (CBT), in particular the intervention termed exposure therapy, as the treatment of choice. However, although the evidence base for cognitive‐behaviour therapy is extensive, the service delivery evidence base is poor, and alternative ways of delivering therapy are required if mental health services are to achieve standards set out by the National Service Framework in the United Kingdom.Aim. The study had two aims: (1) to develop a self‐help manual, which could be facilitated by a nurse trained in CBT, for clients suffering from agoraphobia and (2) to pilot the self‐help manual and evaluate its effectiveness.Method. The self‐help manual was piloted with experienced nurses trained in CBT on three clinical sites for 10 weekly sessions of 30 minutes duration. A range of clinical outcome measures was administered by an independent assessor before and after treatment and at 1‐month follow‐up.Results. A total of 18 clients completed treatment and results showed improvement on all clinical measures; improvement was maintained at 1‐month follow‐up. Importantly, 89% of clients were clinically significantly improved at post‐treatment assessment. Clients were satisfied with their treatment and the self‐help manual, and therapists found facilitated self‐help an acceptable way to deliver treatment.Conclusion. Nurses can deliver effective support to patients using a self‐help manual for agoraphobia. Although the results are promising, further work is required with larger numbers, longer follow‐up and economic evaluation under controlled conditions. The work could also be adapted to different psychological conditions. Variation in the amount of specialist educational training is necessary to determine how many nurses are needed to support patients using self‐help.
Light‐regulated expression of the pea plastocyanin gene is mediated by elements within the transcribed region of the gene
Authors: Helliwell Chris A. , Webster Carl I. , Gray John C. 
Published: August 2003

SummaryExpression of the pea plastocyanin gene (PetE) is regulated by light in both pea and transgenic tobacco plants. However, the PetE promoter with the 5′ untranslated leader region does not direct light‐regulated expression of the GUS reporter gene in transgenic tobacco. This suggested that sequences downstream of the translation start of the PetE gene are required for light‐regulated expression. To investigate this possibility the expression of a series of chimeric gene constructs in transgenic tobacco plants was examined to assess the contributions of the promoter, the 5′ untranslated leader region, the coding region and the 3′ region of the PetE gene to light‐regulated expression. Both the coding region and the 5′ untranslated leader region of the PetE gene were found to be required for full light regulation. Full light regulation of chimeric gene constructs containing the cauliflower mosaic virus (CaMV) 35S promoter required the deletion of CaMV 5′ leader and polylinker sequences from the constructs. The presence of CaMV and polylinker sequences at the 5′ end of the PetE leader masked the light regulation directed by the transcribed region of the pea PetE gene.
Ca2+‐independent phosphorylation of a 68 kDa pollen protein is stimulated by the self‐incompatibility response in Papaver rhoeas
Authors: Rudd Jason J. , Franklin F. Christopher H. , Franklin‐Tong Vernonica E. 
Published: August 2003

SummaryThe self‐incompatibility (SI) response in Papaver rhoeas involves a Ca2+‐based signalling pathway, which mediates the SI‐specific inhibition of incompatible pollen. We have previously reported the identification of p26.1, a pollen protein whose phosphorylation was increased specifically as a consequence of the SI response. We have investigated whether further specific protein phosphorylation events are induced in P. rhoeas pollen. Here we report the identification of an additional pollen protein, p68, which also responds to S proteins by an increase in its phosphorylation state. This phosphorylation event occurs in living pollen tubes grown in vitro, and can be observed specifically when pollen is challenged with biologically active S proteins that are incompatible with the S alleles carried by the pollen and not when pollen was challenged with compatible S proteins. The timing of the increase in phosphorylation of p68 is temporally later than that of p26.1, occurring between 240 sec and 400 sec after challenge. This suggests that its phosphorylation is downstream of p26.1 in the SI signalling pathway(s). Surprisingly, the kinases responsible for the phosphorylation of p68 are not Ca2+‐dependent. This, and the later timing of the p68 response, suggests that a ‘second wave’ of Ca2+‐independent signalling may follow the initial Ca2+‐dependent SI signalling. This indicates that the SI signalling pathway(s) in pollen may be quite complex.
Conserved expression of a TASSELSEED2 homolog in the tapetum of the dioecious Silene latifolia and Arabidopsis thaliana
Authors: Lebel‐Hardenack Sabine , Ye De , Koutnikova Hana , Saedler Heinz , Grant Sarah R. 
Published: August 2003

SummaryTo investigate the genetics of male sex determination and stamen development in the dioecious plant Silene latifolia (white campion), male‐specific transcripts were isolated from developing flowers by cDNA subtraction. One of the cDNAs identified, STA1, had high DNA and amino acid sequence homology to the male sex determining gene of Zea mays (maize), TASSELSEED2. Both genes are expressed in male and not in female flowers, However, they do not share the same expression pattern. The TASSELSEED2 gene product is expressed in the gynoecium primordia of male maize flowers where it is necessary for pistil abortion. STA1 is not expressed in the gynoecium primordia of male white campion and therefore its gene product cannot perform the same function in sex determination that TASSELSEED2 performs in maize. STA1 is expressed in tapetal cells of white campion male flowers and of white campion hermaphroditic mutants. A homologous gene is also expressed in the tapetum of hermaphroditic Silene species. Tapetal expression of a homologous gene (named ATA1) was also found in Arabidopsis thaliana. The similarity in primary sequence and expression pattern of STA1 and ATA1 indicate that these genes have a conserved role in tapetum development.
Systemic elevation of phosphatidic acid and lysophospholipid levels in wounded plants
Authors: Lee Sumin , Suh Sujeoung , Kim Seju , Crain Richard C. , Kwak June Myoung , Nam Hong‐Gil , Lee Youngsook 
Published: August 2003

SummaryPlants develop systemic defense responses upon exposure to pathogens or wounding by herbivores. Lipids and lipid metabolites have previously been implicated in induction of defense molecules during plant responses to physical wounding. Possible involvement of changes in lipid composition in systemic wound signal transduction was examined in leaves of seedlings of several different plant species. In the wounded tomato leaf, phosphatidic acid increased approximately fourfold within 5 min whereas lysophosphatidylcholine and lysophosphatidylethanolamine increased over twofold within 15 min of wounding. Similar changes in these lipids were observed in the neighboring non‐wounded leaf. In broad bean, soybean, sunflower and pepper seedlings phosphatidic acid levels increased rapidly and systemically upon wounding. The results suggest that the role of phospholipid hydrolysis and accumulation of lipid metabolites in the early events are responsible for systemic wound signal transduction in plants. Furthermore, they indicate that the wound signal propagates outside the wounded leaf within 5 min in these plants.
The Arabidopsis MALE STERILITY 2 protein shares similarity with reductases in elongation/condensation complexes
Authors: Aarts Mark G.M. , Hodge Rachel , Kalantidis Kriton , Florack Dion , Wilson Zoe A. , Mulligan Bernard J. , Stiekema Willem J. , Scott Rod , Pereira Andy 
Published: August 2003

SummaryThe Arabidopsis thaliana MALE STERILITY 2 (MS2) gene product is involved in male gametogenesis. The first abnormalities in pollen development of ms2 mutants are seen at the stage in microsporogenesis when microspores are released from tetrads. Expression of the MS2 gene is observed in tapetum of wild‐type flowers at, and shortly after, the release of microspores from tetrads. The MS2 promoter controls GUS expression at a comparable stage in the tapetum of transgenic tobacco containing an MS2 promoter‐GUS fusion. The occasional pollen grains produced by mutant ms2 plants have very thin pollen walls. They are also sensitive to acetolysis treatment, which is a test for the presence of an exine layer. The MS2 gene product shows sequence similarity to a jojoba protein that converts wax fatty acids to fatty alcohols. A possible function of the MS2 protein as a fatty acyl reductase in the formation of pollen wall substances is discussed.
Light‐dependent regulation of carotenoid biosynthesis occurs at the level of phytoene synthase expression and is mediated by phytochrome in Sinapis alba and Arabidopsis thaliana seedlings
Authors: von Lintig Johannes , Welsch Ralf , Bonk Michael , Giuliano Giovanni , Batschauer Alfred , Kleinig Hans 
Published: August 2003

SummaryIn chloroplasts, carotenoids are essential pigments involved in photosynthesis. During photomorphogenesis, a coordinated increase in the amounts of chlorophylls and carotenoids, in conjugation with other components, leads to the formation of a functional photosynthetic apparatus. To investigate the regulation of carotenoid biosynthesis during this process at the molecular level, GGPS, PSY and PDS cDNAs have been cloned from white mustard (Sinapis alba L.). GGPS encodes a key enzyme in plastid isoprenoid metabolism, while the products of PSY and PDS catalyse the subsequent steps in carotenoid biosynthesis. Due to the low mRNA levels of the genes involved, the use of a RT‐PCR protocol was necessary to measure gene expression during photomorphogenesis. With light, there is an up‐regulation of PSY expression, the first gene within the carotenoid biosynthetic pathway, while PDS and GGPS expression levels remain constant. Treatment with different light qualities reveals a phytochrome‐mediated regulation of PSY expression in developing white mustard seedlings. To obtain more detailed information on the light‐regulation, Arabidopsis thaliana wild‐type and phytochrome mutants were utilized. Continuous far‐red and red light both increase the expression of PSY in wild‐type seedlings, demonstrating that both light‐labile and light‐stable phytochromes are involved in PSY regulation. The response to far‐red light is completely abolished in the phyA mutant, showing that PHYA mediates the increase in PSY transcript levels under these light conditions. In the phyB mutant, the red light response is normal, indicating that PSY expression is not controlled by PHYB but by other light‐stable phytochromes. Measurement of chlorophylls and carotenoids under the same light regimes shows that the up‐regulation of PSY expression does not necessarily result in an increase of the carotenoid content. Only those light conditions which allow chlorophyll biosynthesis lead to a significant increase of the carotenoid content. Therefore, it is proposed that up‐regulation of PSY mRNA levels leads to an increased capacity for the formation of carotenoids. However, this only takes place under light conditions leading to protochlorophyllide photoconversion.
Developmental and auxin‐induced expression of the Arabidopsis prha homeobox gene
Authors: Plesch Gunnar , Störmann Katja , Torres Jorge Tovar , Walden Richard , Somssich Imre E. 
Published: August 2003

SummaryA single‐copy Arabidopsis homeobox gene, prha, which encodes a homeodomain protein with a molecular weight of 90 500 has been characterized. The position of the gene was mapped to the distal part of chromosome 4. Expression of the gene differs in various vegetative and floral plant tissues and is positively influenced by the phytohormone auxin. In Arabidopsis plants, a complex pattern of prha promoter‐driven GUS expression is observed, often associated with regions of developing vascular tissue. Exogenously applied auxin strongly increased endogenous prha transcript levels. In addition, the prha promoter is highly responsive to the synthetic auxin, naphthalene acetic acid, in transient assays using tobacco protoplasts. The PRHA protein has the capacity to bind to TAATTG core sequence elements but requires additional adjacent bases for high‐affinity binding. These findings are discussed in relation to studies of other plant homeobox genes as well as possible in vivo target genes for PRHA.
Overexpression of light‐dependent PORA or PORB in plants depleted of endogenous POR by far‐red light enhances seedling survival in white light and protects against photooxidative damage
Authors: Sperling Ulrich , van Cleve Barbara , Frick Geneviève , Apel Klaus , Armstrong Gregory A. 
Published: August 2003

SummaryThe structurally related light‐dependent protochlorophyllide (Pchlide) oxidoreductases PORA and PORB mediate the only light‐requiring step in chlorophyll (Chl) biosynthesis in higher plants. Correlative evidence suggests that some in vivo functions of PORA and PORB may be unique, including a postulated photoprotective role for PORA. For example, wild‐type Arabidopsis thaliana seedlings grown in non‐photooxidative far‐red light (cFR) resemble those grown in white light (WL), but they are yellow and do not green normally thereafter in WL. This defect is accompanied by the absence of detectable PORA and reduced levels of PORB expression. Here, direct evidence is provided that the presence of POR, either as PORA or PORB, can confer photoprotection in plants. In contrast to the wild‐type, the plastids of transgenic PORA‐ or PORB‐overexpressing Arabidopsis seedlings grown in cFR possess extensive prolamellar bodies. Upon a subsequent shift to WL, POR‐overexpressing seedlings develop thylakoid membranes, accumulate large amounts of Chl and are viable at fluence rates lethal to the wild‐type. Intriguingly, the plastid membrane architectures of greening transgenic seedlings seem to depend on whether PORA or PORB has been over‐produced. POR‐overexpressing seedlings shifted from cFR to WL of fluence rates from 20 to 500 μE m−2 sec−1 accumulate substantially higher amounts of Chl than does the wild‐type. Furthermore, the WL fluence rate that permits maximal Chl accumulation increases from 8 μE m−2 sec−1 in the wild‐type to 125 μE m−2 sec−1 in transgenic seedlings. POR Overexpression during growth in cFR also correlates with a fourfold decrease in the steady‐state content of Pchlide, a potentially lethal photosensitizer.
The VLF loci, polymorphic between ecotypes Landsberg erecta and Columbia, dissect two branches of phytochrome A signal transduction that correspond to very‐low‐fluence and high‐irradiance responses
Authors: Yanovsky Marcelo J. , Casal Jorge J. , Luppi Juan P. 
Published: August 2003

SummaryPhytochromes play a key role in the perception of light signals by plants. In this study, the three classical phytochrome action modes, i.e. very‐low‐fluence responses (VLFR), low‐fluence responses (LFR) and high‐irradiance responses (HIR), were genetically dissected using phyA and phyB mutants of Arabidopsis thaliana (respectively lacking phytochrome A or phytochrome B) and a poly‐morphism between ecotypes Landsberg erecta and Columbia. Seed germination and potentiation of greening, hypocotyl growth inhibition and cotyledon unfolding in etiolated seedlings of the ecotype Landsberg erecta showed biphasic responses to the calculated proportion of active phytochrome established by one light pulse or repeated light pulses. The first phase, i.e. the VLFR, was absent in the phyA mutant, normal in the phyB mutant (both in the Landsberg erecta background) and severely deficient in Columbia. The second phase, i.e. the LFR, was present in the phyA mutant, deficient in the phyB mutant and normal in Columbia. Under continuous far‐red light, HIR of etiolated seedlings were absent in phyA and normal in phyB and Columbia. The segregation of VLFR in recombinant inbred lines derived from a cross between Landsberg erecta and Columbia was analysed by mapmaker/QTL. Two quantitative trait loci, one on chromosome 2 (VLF1) and another on chromosome 5 (VLF2), were identified as responsible for the polymorphism. Phytochrome A is proposed to initiate two transduction pathways, VLFR and HIR, involving different cells and/or different molecular steps. This is the first application of the analysis of quantitative trait loci polymorphic between ecotypes to dissect transduction chains of environmental signals.
Enhanced tolerance to bacterial pathogens caused by the transgenic expression of barley lipid transfer protein LTP2
Authors: Molina A. , García‐Olmedo F. 
Published: August 2003

SummaryPurified lipid transfer protein LTP2 from barley applied on tobacco leaves eliminated symptoms caused by infiltration of Pseudomonas syringae pv. tabaci 153. Growth of the pathogen in leaves of transgenic tobacco plants was retarded when compared with non‐transformed controls. The percentage of inoculation points that showed necrotic lesions was greatly reduced in transgenic tobacco (17–38% versus 78%) and the average size of these lesions was 61–81% that of control. The average total lesion area (necrosis and chlorosis) in the transgenic plants was also reduced (38% of control). Arabidopsis thaliana transgenic plants inoculated with P. syringae pv. tomato DC3000 also had lower percentages of necrotic lesions (22–38% versus 76%), a reduced average area for each lesion (53–67% of control), and a smaller total lesion area per inoculation (43% of control). These results further support the assignment of a defense role for LTPs and highlight their biotechnological potential.
Cloning and characterization of LRG5, a gene involved in blue light signaling in Chlamydomonas gametogenesis
Authors: Glöckner Gernot , Beck Christoph F. 
Published: August 2003

SummaryThe Chlamydomonas reinhardtii mutant lsrg2 exhibits a partial defect in the blue‐light activated signal transduction chain that controls gametogenesis. By genomic complementation, a cosmid clone that corrects the Irg2 phenotype was isolated. The smallest fragment of this cosmid that gave complementation was used in RFLP analysis, which revealed that the cloned gene was not linked to the LRG2 locus. The sequence of the genomic clone and a cDNA of this gene, which we named LRG5, was determined. Although no significant homology with sequences in the databases was found, the nuclear localization signal and the presence of negatively and positively charged domains suggests a function for the LRG5 protein in the regulation of transcription. The correction of the Irg2 phenotype is probably caused by overexpression of the LRG5 gene in the transformants, as these expressed a level of LRG5 approximately twofold higher than wild‐type or mutant strains. Southern blot analysis provided evidence for the existence of homologous sequences in other green algae as well as in higher plants, suggesting a conserved function for the LRG5 protein.
Characterization of the targeted nuclear accumulation of GFP within the cells of transgenic plants
Authors: Grebenok Robert J. , Lambert Georgina M. , Galbraith David W. 
Published: August 2003

SummaryThe soluble proteins of the nucleoplasm are synthesized on cytoplasmic ribosomes. Proteins larger than about 40 kDa are post‐translationally targeted to the nucleus via energy‐dependent processes, passing through the nuclear pore complex into the nucleoplasm. Targeting involves nuclear localization signals (NLSs) found within the primary sequences of the imported proteins. In higher plants, information has come primarily from study of proteins carrying ‘classical’ NLSs, comprising stretches of basic amino acids, and has required assays to measure nuclear uptake both in vitro and in vivo. In general, these assays are not entirely satisfactory; they are either technically demanding, are of limited accuracy and statistical rigor, or are unsuitable for in vivo applications.The green‐fluorescent protein (GFP) of Aequorea victoria has recently emerged as a versatile marker for transgenic expression in vivo. Conditions under which GFP gene fusions can be employed for the analysis of nuclear targeting in plant protoplasts have been described. This study demonstrates for the first time the nuclear targeting of chimeric GFP molecules in transgenic tobacco. This is accompanied by a description and evaluation of novel analytical methods, involving flow and image cytometry, for the quantitative temporal and spatial analysis of nuclear targeting, and these unique methods are used to provide information concerning the targeting process. Finally, the way in which the chimeric GFP molecules might be employed for the study of various important problems in plant cell and developmental biology is discussed.
A negative selection scheme based on the expression of cytosine deaminase in plastids
Authors: Serino German , Maliga Pal 
Published: August 2003

SummaryThe enzyme cytosine deaminase (CD) encoded by codA catalyzes deamination of cytosine to uracil. CD is present in prokaryotes and in many eukaryotic micro‐organisms, but is absent in higher plants. 5‐fluorocytosine (5FC) is metabolized in CD‐expressing cells, causing cellular death. A chimeric codA has been introduced into the tobacco plastid genome and 5FC was used to select against tissue culture cells and seedlings expressing CD. This negative selection scheme will be useful in identifying nuclear genes which control plastid gene expression in higher plants.
Construction of an ∼2 Mb contig in the region around 80 cM of Arabidopsis thaliana chromosome 2
Authors: Wang Ming Li , Huang Lu , Bongard‐Pierce Deverie K. , Belmonte Stephen , Zachgo Eve Ann , Morris John W. , Dolan Maureen , Goodman Howard M. 
Published: August 2003

SummaryA method for construction of bacterial artificial chromosome (BAC) contigs from a yeast artifical chromosome (YAC) physical map is described. An ∼2 Mb contig, consisting of two large BAC contigs linked by a small YAC, has been assembled in the region around 80 cM of Arabidopsis thaliana chromosome 2. Clones from this contig will facilitate gene isolation in the region and can be used directly as substrates for DNA sequencing.
Heat sensitivity of human parvovirus B19
Authors: Yunoki M. , Tsujikawa M. , Urayama T. , Sasaki Y. , Morita M. , Tanaka H. , Hattori S. , Takechi K. , Ikuta K. 
Published: August 2003

Background and Objectives To date there has been no published report on a systematic evaluation of the heat sensitivity of human parvovirus B19 (B19) and the related safety of the plasma‐derived fractionated products. In this study, we examined the heat sensitivity of B19 by using the infectivity assay with cultured cells.Materials and Methods The heat sensitivity of B19 was examined by measuring the reduction in viral infectivity titres after heating liquid containing B19 at 60 °C. Viral infectivity was assayed by detection of viral antigens or viral mRNA in infected cells. As a control, canine parvovirus (CPV) was also heat‐treated.Results B19 displayed quite different inactivation kinetics to CPV when both were heated in liquid at 60 °C. In sharp contrast to the latter, B19 was rapidly inactivated within 1 h when the virus was suspended in 5% or 25% human serum albumin solution, phosphate‐buffered saline, or complete medium. However, B19 appeared to be resistant to heat inactivation in liquid containing 60% sucrose.Conclusions The heat sensitivity of B19 in liquid was clearly different from that of CPV. Significantly, the efficiency to inactivate B19 and reduce its infectivity following heating in liquid was mainly affected by the composition of the solutions used for virus suspension.