bluuebunny/crossref_metadata_2025 huggingface dataset:
Mononuclear cells from patients recovered from cutaneous leishmaniasis respond toLeishmania majoramastigote class I nuclease with a predominant Th1-like response
Authors: Farajnia S , Mahboudi F , Ajdari S , Reiner N E , Kariminia A , Alimohammadian M H
Published: February 2005

The Leishmania major amastigote class I nuclease (LmaCIN) is a developmentally regulated protein that is highly expressed in the amastigote stage of L. major. This protein is homologous to the P4 nuclease of L. pifanoi, which has been shown to induce protective immune response in a murine model. To evaluate LmaCIN as a potential human vaccine candidate, cellular immune responses to recombinant LmaCIN were examined in individuals recovered from Old World cutaneous leishmaniasis. Peripheral blood mononuclear cells (PBMC) from patients recovered from L. major infection were cultured either with recombinant LmaCIN or autoclaved L. major (ALM) as control. rLmaCIN induced significant proliferation of PBMC from 90% of recovered patients. Phenotypic analysis of proliferating cells showed that CD8+ cells were the predominant cell type proliferating in response to rLmaC1N. Screening of culture supernatants for cytokines showed that rLmaCIN induced high levels of interferon (IFN)-γ (mean ± s.e.m.: 1398 ± 179 pg/ml) associated with little interleukin (IL)-10 and little or no IL-5 production. These findings show that LmaCIN is immunogenic in humans during L. major infection and that it can elicit immunological responses relevant to immunoprophylaxis of leishmaniasis.
Increased lymphocyte trafficking to colonic microvessels is dependent on MAdCAM-1 and C-C chemokine mLARC/CCL20 in DSS-induced mice colitis
Authors: Teramoto K , Miura S , Tsuzuki Y , Hokari R , Watanabe C , Inamura T , Ogawa T , Hosoe N , Nagata H , Ishii H , Hibi T
Published: February 2005

Although enhanced lymphocyte trafficking is associated with colitis formation, little information about its regulation is available. The aim of this study was to examine how the murine liver and activation-regulated chemokine (mLARC/CCL20) contributes to lymphocyte recruitment in concert with vascular adhesion molecules in murine chronic experimental colitis. T and B lymphocytes isolated from the spleen were fluorescence-labelled and administered to recipient mice. Lymphocyte adhesion to microvessels of the colonic mucosa and submucosa was observed with an intravital microscope. To induce colitis, the mice received two cycles of treatment with 2% dextran sodium sulphate (DSS). In some of the experiments antibodies against the adhesion molecules or anti-mLARC/CCL20 were administered, or CC chemokine receptor 6 (CCR6) of the lymphocytes was desensitized with excess amounts of mLARC/CCL20. Significant increases in T and B cell adhesion to the microvessels of the DSS-treated mucosa and submucosa were observed. In chronic colitis, the accumulation of lymphocytes was significantly inhibited by anti-mucosal addressin cell adhesion molecule (MAdCAM)-1 mAb, but not by anti-vascular cell adhesion molecule-1. In DSS-treated colonic tissue, the expression of mLARC/CCL20 was significantly increased, the blocking of mLARC/CCL20 by monoclonal antibody or the desensitization of CCR6 with mLARC/CCL20 significantly attenuated the DSS-induced T and B cell accumulation. However, the combination of blocking CCR6 with MAdCAM-1 did not further inhibit these accumulations. These results suggest that in chronic DSS-induced colitis, both MAdCAM-1 and mLARC/CCL20 may play important roles in T and B lymphocyte adhesion in the inflamed colon under flow conditions.
Characterization of human peritoneal dendritic cell precursors and their involvement in peritonitis
Authors: McCully M L , Chau T A , Luke P , Blake P G , Madrenas J
Published: February 2005

Scattered evidence suggests that the human peritoneal cavity contains cells of the dendritic cell (DC) lineage but their characterization is missing. Here, we report that the peritoneal cavity of normal subjects and of stable patients on peritoneal dialysis (PD) contains a population of CD14+ cells that can differentiate into DCs or macrophages. Within this pool, we characterized a CD14+CD4+ cell subset (2.2% of the peritoneal cells) fulfilling the definition of myeloid DC precursors or pre-DC1 cells. These cells expressed high levels of HLA-DR, CD13, CD33, and CD86, and low levels of CD40, CD80, CD83, CD123, CD209, TLR-2 and TLR-4. These cells retained CD14 expression until late stages of differentiation, despite concomitant up-regulation of DC-SIGN (CD209), CD1a, CD80 and CD40. Peritoneal pre-DC1 cells had endocytic capacity that was down-regulated upon LPS/IFN-γ stimulation, were more potent allo-stimulators than peritoneal CD14+CD4–/lo cells and monocyte-derived macrophages, and induced Th1 cytokine responses. More importantly, the number of peritoneal pre-DC1 cells increased during PD-associated peritonitis, with a different profile for Gram positive and Gram negative peritonitis, suggesting that these cells participate in the induction of peritoneal adaptive immune responses, and may be responsible for the bias towards Th1 responses during peritonitis.
Rituxan (anti-CD20 antibody)-induced translocation of CD20 into lipid rafts is crucial for calcium influx and apoptosis
Authors: Janas E , Priest R , Wilde J I , White J H , Malhotra R
Published: February 2005

Rituxan, a chimeric anti-CD20 antibody, is the first antibody approved for immunotherapy in non-Hodgkin's B-cell lymphoma and other B-cell lymphoproliferative disorders. Additionally, efficacy of Rituxan treatment has been reported in nonmalignant autoimmune diseases such as rheumatoid arthritis. Crosslinking of CD20 molecules by Rituxan induces therapeutic B-cell depletion. CD20 is a B-lymphocyte specific integral membrane protein, proposed to function as a store-operated calcium channel, which is activated upon receptor-stimulated calcium depletion of intracellular stores. Crosslinking of CD20 by antibodies has been reported to induce a redistribution of CD20 molecules to specialized microdomains at the plasma membrane known as lipid rafts. Here, we report that in the absence of Rituxan, CD20 exhibits a low affinity to lipid rafts. However, binding of Rituxan significantly increases the affinity of CD20 for lipid rafts resulting in its redistribution to a fraction resistant to Triton X-100 solubilization. Furthermore, we demonstrate that disturbing the raft integrity by cholesterol extraction results in dissociation of CD20 from a Triton X-100 resistant fraction followed by complete inhibition of Rituxan-induced calcium entry and apoptosis. The integrity of lipid rafts seems to play a crucial role for CD20-induced caspase activation. These data show, for the first time, that Rituxan-induced translocation of CD20 to lipid rafts is important for increased intracellular Ca2+ levels and downstream apoptotic signalling.
Striatal Dopamine D1 receptors in narcolepsy: A PET study with [11C]NNC 756
Authors: RINNE J. O. , HUBLIN C. , PARTINEN M. , RUOTTINEN H. , NÅGREN K. , LEHIKOINEN P. , RUOTSALAINEN U. , LAIHINEN A.
Published: February 2005

SUMMARY We investigated dopamine D, receptors in the putamen and caudate nucleus with positron emission tomography in six patients with narcolepsy and five healthy controls using [11C]NNC 756 as ligand. The caudate‐to‐cerebellum and putamen‐to‐cerebellum ratios of [11C]NNC 756 were within normal limits in patients with narcolepsy. No evidence of increased D1, receptor binding in narcolepsy was found.
Work hours, sleep patterns and fatigue among merchant marine personnel
Authors: SANQUIST THOMAS F. , RABY MIREILLE , FORSYTHE ALICE , CARVALHAIS ANTONIO B.
Published: February 2005

SUMMARY A field study of work and sleep patterns among commercial merchant marine personnel is reported. Data collected over a 10‐30‐d period from 141 subjects aboard eight ships included information concerning work‐rest schedules, sleep timing, alertness on the job and critical fatigue. The data indicate that watchstanders on the 4‐on, 8‐off schedule show considerable disruption in their sleep. The average sleep duration for all mariners is 6.6 h; watchstanders obtain their sleep in fragmented periods that are frequently less than 5 h in duration. Analysis of critical fatigue shows an incidence of 1‐24% across personnel and measures. Of particular concern are the watchstanders on the 04.00‐08.00 schedule, who sleep less than 4 h per 24‐h period 22% of the time. Potential countermeasures, including changes in scheduling and staffing are proposed.
Organ‐dependent regulation of a plant promoter isolated from rice by ‘promoter‐trapping’ in tobacco
Authors: Claes B. , Smalle J. , Dekeyser R. , Van Montagu M. , Caplan A.
Published: February 2005

A vector containing a transcriptionally inactive neomycin phosphotransferase II gene was used to select promoter sequences from a pool of random genomic DNA fragments. This paper describes how one such sequence (P4.7) isolated from Oryza sativa acts as a hormonally regulated promoter in Nicotiana tabacum. Relative expression ratios in leaf, root, midrib, callus, and stem tissue of tobacco plants are 1:5:4:10:17. Histochemical assays show that P4.7 activates the uidA reporter gene throughout the phloem and cortex of tobacco stems. Transcription from the P4.7 fragment is inducible in leaf tissue by low levels of a‐naphthalene acetic acid or 6–benzyl‐aminopurine, even when cell proliferation is inhibited by colchicine or hydroxyurea. Conversely, 1 % DMSO was found to inhibit activation of P4.7 without interfering with callus formation. The fragment contains TATA and CAAT sequences normally found at the 5′ end of many plant genes, and an additional region homologous to sequences located in similar positions in a variety of similarly regulated promoters. Promoter deletion and fusion experiments have indicated the location of a stem enhancer element in P4.7. The promoter trap system we have described may potentially be used to characterize transcriptional factors common to monocot and dicot species.
T‐DNA insertion mutagenesis in Arabidopsis: mutational spectrum
Authors: Feldmann Kenneth A.
Published: February 2005

More than 8000 transformants of Arabidopsis have been generated by treating germinating seeds with cultures of Agrobacterium tumefaciens. Genetic characterization of a subset of the transformants indicates that they contain an average of 1.4 inserts each, as assayed by kanamycin resistance. Molecular analysis shows that the inserts are predominantly concatamers of T‐DNAs arranged as direct and inverted repeats. More recently these 8000 lines have been screened under a variety of growth conditions for visible alterations in phenotype. More than 1000 putative mutants were observed during the application of these screening procedures. These mutants fall into several general classes: seedling‐lethals, size variants, pigment, embryo‐defective, reduced‐fertility, dramatic (morphological), and physiological. The majority of the mutants (88%) segregate in a Mendelian manner for the mutant phenotype. An analysis of approximately 50 mutants in this group shows that > 80% are tagged with a functional insert. The wide spectrum of mutants observed suggests that it may be feasible to develop a comprehensive collection of mutant lines in which each gene is tagged by a T‐DNA insertion.
Identification of a novel S‐phase‐specific gene during the cell cycle in synchronous cultures of Catharanthus roseus cells
Authors: Ito Masaki , Kodama Hiroaki , Komamine Atsushi
Published: February 2005

The cell‐cycle specific cDNAs were isolated from a cDNA library prepared from cells in the S phase in the synchronous cultures of Catharanthus roseus. One of the isolated genes, which we refer to as cyc07, was analyzed in detail. The full‐length cDNA of cyc07 contains an open reading frame of 735 nucleotides, encoding a protein of 245 amino acids with a molecular weight of 28356 Da. The protein predicted from the nucleotide sequence is highly basic, as are mammalian histones. cyc07 mRNA was detected specifically in cells at the S phase in synchronous cultures. The induction and accumulation of mRNA in the S phase were suppressed when DMA synthesis was inhibited by aphidicolin. In the intact plant, cyc07 mRNA was found preferentially in root tips that contained meristematic tissue. A databank search revealed that a sequence homologous to the nucleotide sequence of cyc07 cDNA is present in the downstream region of the SIR3 gene in the yeast genome. The amino acid sequence predicted from the corresponding region of the yeast genome exhibited significant homology with that of cyc07 protein. These similarities between cyc07 and the corresponding region in yeast suggest that the homologous sequence in yeast is a novel gene that is functionally homologous to cyc07. Our results presented here suggest the possibility that cyc07 may play a role in the proliferation of higher plant cells, in particular in the entry into or progression of the S phase of the cell cycle.
Mitochondrial genome organization and expression associated with cytoplasmic male sterility in sunflower (Helianthus annuus)
Authors: Laver H.K. , Reynolds S.J. , Moneger F. , Leaver C.J.
Published: February 2005

A comparative investigation of the organization and expression of the mitochondrial genome in fertile and cytoplasmic male sterile (CMS) sunflower (Helianthus annuus) has been undertaken. A region of mitochondrial genome variation between the two phenotypes has been located in the 3′ flanking region of the gene encoding the α subunit of the F1 ATPase (atpA). Physical mapping and sequence analysis have been used to show that a rearrangement involving an inversion and an insertion has occurred immediately downstream of the atpA coding region in the mitochondrial DMA from sterile sunflower. This rearrangement has resulted in the creation of a new open reading frame (ORFc) which is co‐transcribed with atpA in sterile sunflower. In organello labelling of mitochondrial translation products from the two types of sunflower shows that a 15 kDa protein is synthesized by the mitochondria from sterile sunflower but not by those from fertile plants. The ORFc sequence could encode this 15 kDa protein which may be causally related to the CMS phenotype.
A promoter sequence involved in cell‐specific expression of the pea glutamine synthetase GS3A gene in organs of transgenic tobacco and alfalfa
Authors: Brears Timothy , Walker Elsbeth L. , Coruzzi Gloria M.
Published: February 2005

The DMA sequence of the pea cytosolic glutamine synthetase GS3A gene promoter has been determined and the start of transcription mapped using S1 nuclease. The full‐length promoter and a series of 5′ deletions were fused to β‐glucuronidase (GUS) and introduced into transgenic tobacco and alfalfa. In transgenic tobacco the GS3A promoter directed GUS expression in the phloem cells of the vasculature in leaves, stems and roots. GUS expression was also detected in the vasculature of cotyledons and the root tips of germinating T1 seedlings. The promoter conferred a similar expression pattern in transgenic alfalfa, and expression was also observed in root nodules. Nodule expression was located in nodule primordia, as well as the meristem, symbiotic zone, and vasculature of mature nodules. The promoter was found to be active even when deleted to ‐132 relative to the start of transcription. DMA mobility‐shift analysis identified a protein present in nuclear and whole‐cell plant extracts which bound to a 17 bp DNA element contained within the minimal ‐132 promoter required for expression.
The MADS box gene family in tomato: temporal expression during floral development, conserved secondary structures and homology with homeotic genes from Antirrhinum and Arabidopsis
Authors: Pnueli L. , Abu‐Abeid M. , Zamir D. , Nacken W. , Schwarz‐Sommer Zs. , Lifschitz E.
Published: February 2005

Five genes with homology to the floral homeotic genes deficiens of Antirrhinum and agamous of Arabidopsis were isolated from tomato. Each of the five genes is unique in the genome and could be localized to a different chromosome by RFLP mapping. Four of the tomato genes (hereafter TM) are flower‐specific with distinguishable temporal expression. TM4 and TM8 are ‘early’, while TM5 and TM6 are ‘late’ genes. TM4 is homologous to squamosa and TM6 is similar to deficiens, which are, respectively, ‘early’ and ‘late’ bona fide homeotic genes in Antirrhinum. The proteins encoded by the five tomato genes, like several known homeotic genes from other plants, contain within their N‐terminus a highly conserved DMA‐binding domain, the MADS box. All known plant MADS box genes also share, however, other properties. They all contain a central, moderately conserved, and rather basic domain, and a highly divergent or even missing C‐terminal domain. Furthermore, molecular modelling predicts the presence of a conserved amphipatic alpha helix, at a constant distance from the MADS box in each of these proteins. The common properties of eight MADS box proteins from three plant families indicate that all their domains were coded for by the same ancestor gene. The sequence homology between pairs of MADS genes from different species indicates that the MADS ancestor gene multiplied and diverged in an ancestor plant common to several dicotyledon families.
In‐situ enzyme histochemistry on plastic‐embedded plant material. The development of an artefact‐free β‐glucuronidase assay
Authors: De Block Marc , Debrouwer Dirk
Published: February 2005

An in‐situ enzyme histochemical method is described that preserves the tissue and cell structure as well as the enzyme activities. Flower buds at different developmental stages from wild‐type and transgenic Brassica napus plants, the latter containing the GUS gene under the control of a tapetum‐specific promoter, were used as starting material. The method is based on the following principles: processing of the tissue on crushed ice, no fixation but a pretreatment with spermidine, partial dehydration with acetone, and a final embedding in a water‐miscible glycol methacrylate resin at 5°C. This method was used to set up a sensitive p‐glucuronidase histochemical assay with a high resolution. A succinate dehydrogenase assay was included as a control for tissue and cell viability as well as a standard for the relative metabolic activities between different tissues and cell types.
A 126 bp fragment of a plant histone gene promoter confers preferential expression in meristems of transgenic Arabidopsis
Authors: Atanassova Rossitza , Chaubet Nicole , Gigot Claude
Published: February 2005

The tissue‐specific pattern of expression directed by the H4A748 Arabidopsis histone promoter was investigated by analysis of β‐glucuronidase (GUS) activity in transgenic Arabidopsis containing H4A748‐GUS gene fusions. As determined by fluorimetric and histochemical tests, the H4A748 promoter directs preferential expression in meristems of young seedlings and adult plants. The low activity found in non‐proliferating tissues may relate to basal constitutive expression of the histone promoter and/or to endo‐reduplication occurring in some tissues. The endogenous histone mRNA levels parallel the GUS activity found in different tissues.

Analysis of the regulatory properties of 5′ deleted promoters showed that multiple positive elements exist between −900 and −219 and that the proximal region of the promoter to −219 is sufficient to establish the full tissue‐specific pattern of expression. Further deletion to −93 nearly abolished the promoter activity thus suggesting that the 126 bp fragment located between −219 and −93 contains the elements responsible for the specific expression pattern. The presence of several remarkable sequences within this fragment is discussed.
Artifacts in the localization of GUS activity in anothers of petunia transformed with a CaMV 35S‐GUS construct
Authors: Mascarenhas Joseph P. , Hamilton Douglas A.
Published: February 2005

Blue staining of pollen in transgenic petunia plants containing a β‐glucuronidase gene under the control of a cauliflower mosaic virus 35S promoter, is an artifact caused by the diffusion of reaction products from sites in the another and their dimerization into indigo in pollen. The CaMV 35S promoter is active in various tissues of the another. When pollens from Tradescantia and snapdragon are mixed with transgenic petunia anothers during staining they stain blue, whereas they are colorless when stained alone. In assays for promoter activity in pollen it is essential that pollen is isolated from the another or another section prior to staining for GUS activity, to prevent diffusion artifacts and false staining of pollen.
Temporal and spatial control of expression of anthocyanin biosynthetic genes in developing flowers of Antirrhinum majus
Authors: Jackson David , Roberts Keith , Martin Cathie
Published: February 2005

The co‐ordination of expression of anthocyanin biosynthetic genes was studied in developing flowers. Four genes encoding enzymes operating late in the anthocyanin biosynthetic pathway are induced together during flower development but the early steps appear to be induced more rapidly. Co‐ordination of expression could imply a common regulatory mechanism controlling the expression of metabolically related genes. The data presented here show that while four genes may share such a mechanism for the control of their expression during flower development, different control processes regulate the early steps of the pathway. Spatially, gene expression is patterned across the flower and appears to be very similar for all the biosynthetic genes. However, the observed influence of the regulatory gene Delila shows that the spatial co‐ordination of gene expression must involve more than one regulatory system. Delila itself appears to have a dual function, being required for activation of expression of the later genes in the flower tube but repressing chalcone synthase gene expression in the mesophyll of the corolla lobes. It is postulated that common signals induce the expression of genes in the pathway during flower development. The data presented here suggest that the same regulatory mechanism interprets these signals for four of the genes encoding the later biosynthetic enzymes, but that different or modified mechanisms interpret the signals to control expression of chalcone synthase and chalcone isomerase genes in Antirrhinum flowers.
Tobacco lines with high copy number of replicating recombinant geminivirus vectors after biolistic DNA delivery
Authors: Kanevski Ivan F. , Thakur Sanjay , Cosowsky Laurey , Sunter Garry , Brough Clare , Bisaro David , Maliga Pal
Published: February 2005

The feasibility of obtaining clonal lines with replicating, multicopy geminivirus vectors by direct DNA trans‐formation of cultured tobacco cells was studied. The replicating vectors pTGA32 and pST31 are based on the tomato golden mosaic virus (TGMV) A genome and encode the neomycin phosphotransferase type II (NPT‐II) enzyme that confers kanamycin resistance to plant cells. Following introduction into plant cells, unit‐length viral genomes were released from the tandem repeats and replicated. In protoplasts, replication of unit‐length pTGA32 and pST31 was about as efficient as replication of unit‐length DNA A from plasmid pTGA26, which contains 1.5 copies of wild‐type DNA A. Tobacco suspension culture cells were bombarded with the recombinant DNA A constructs and selected for kanamycin resistance. The number of kanamycin‐resistant clones per bombardment was about the same when the TGMV DNA A vectors or a non‐replicating plasmid (pLC14) which also encodes NPT‐II was used. Replicating, unit‐length DNA A in up to approximately 1000 copies per cell was found in about 10% of the kanamycin‐resistant clones selected following bombardment of cells with TGMV vectors. The results suggest that geminiviruses may serve as useful multicopy vectors in cultured cells.
A genetically definedtrans‐acting locus regulatesS‐locus function inBrassica
Authors: Nasrallah Mikhail E. , Kandasamy Muthugapatti K. , Nasrallah June B.
Published: February 2005

A new locus,SCF1, which regulatesS‐locus function has been identified inBrassica campestris. The spontaneousscf1mutation described in this paper causes the stigma to be receptive to all pollen including selfpollen, but does not affect the pollination phenotype of the male gametophyte. In the stigmas ofscf1homozygotes, theS‐locus‐linkedSLGgene and two other unlinked members of theS‐multigene family, all of which encode secreted glycoproteins, are coordinately down‐regulated at the RNA level, while the expression of theS‐locus‐derivedSRKgene, which encodes a putative receptor protein kinase, is unaffected. It is proposed that theSCF1locus encodes a positivetrans‐acting factor required for the enhanced transcription of several genes in the stigmatic papillar cells during pistil maturation. The effects of thescf1mutation provide strong support for the involvement of the clonedS‐genes in the operation of the pollen‐stigma interaction of incompatibility inB. campestrisand related crucifer species.
Developmental control of the β‐phaseolin gene requires positive, negative, and temporal seed‐specific transcriptional regulatory elements and a negative element for stem and root expression
Authors: Burow Mark D. , Sen Partha , Chlan Caryl A. , Murai Norimoto
Published: February 2005

To identify cis‐acting regulatory elements responsible for developmental control of the common bean seed storage protein β‐phaseolin, a series of 5′‐deletion mutants of the 782 bp upstream sequence together with the coding and 3′‐regions of the β‐phaseolin gene were used to transform tobacco. One major positive regulatory element (−295/−228) and a putative minimal promoter (−64/−14) were indicated by large reductions in phaseolin mRNA and protein concentrations in seeds of plants deficient for these regions. In addition, minor negative (−422/−296) and positive (−782/−423) elements also influenced the level of phaseolin mRNA expression in seeds. One temporal element (−295/−107) governed late expression of phaseolin mRNA in seeds, and possibly a second (−64/−14) regulated early expression. The −64/−14 promoter containing two TATA boxes conferred spatially regulated gene expression, and was sufficient for a low level of expression in root and stem. Significant levels of phaseolin mRNA and protein were detected in stem cortices and secondary roots of plants lacking the −295/−107 negative element. No significant expression in leaf tissue was detected. Results demonstrate that developmental control of β‐phaseolin requires a minimal promoter, one element for the suppression of expression in root and stem tissue, three elements governing quantitative expression in seeds, and at least one temporal element controlling expression in seeds.
Characterization of a new class of oleosins suggests a male gametophyte specific lipid storage pathway
Authors: Roberts Michael R. , Hodge Rachel , Ross Jo H.E. , Sorensen Anna , Murphy Denis J. , Draper John , Scott Rod
Published: February 2005

The expression and sequences of two related anther‐specific cDNAs (13 and C98) of Brassica napus (oilseed rape) were examined. These cDNAs were found to exhibit significant predicted amino acid sequence similarity with a group of seed‐specific proteins, the oleosins, which are involved in oil body membrane structure. Pollen of B. napus also contains oil bodies, and the synthesis and accumulation of these organelles correlates with expression of the 13 and C98 transcripts. The protein content of purified pollen oil bodies was therefore examined in order to determine the presence or absence of possible oleosins. One major protein species of 14 kDa was identified and subjected to N‐terminal amino acid sequence determination. The sequence of this pollen oil body associated protein is homologous to the predicted sequence of one of the anther‐specific cDNAs, 13, implying that this cDNA represents an anther‐specific oleosin gene expressed in developing pollen. The expression of other genes active during seed oil body formation was also examined, and these were found to be specific to the sporophyte, inferring the existence of specific components of the pathway of lipid synthesis and storage in the male gametophyte. These possibilities and the relationship between different oleosin genes are discussed.
Seed‐specific expression of a bacterial desensitized aspartate kinase increases the production of seed threonine and methionine in transgenic tobacco
Authors: Karchi Hagai , Shaul Orit , Galili Gad
Published: February 2005

In order to study the regulation of threonine and methionine synthesis in plant seeds, tobacco plants were transformed with a chimeric gene containing the coding DNA sequence of a mutant lysC gene from Escherichia coli fused to a promoter from a phaseolin seed storage protein gene. The bacterial mutant lysC gene codes for aspartate kinase (AK) which is desensitized to feedback inhibition by lysine and threonine. Increased AK activity, compared with control non‐transformed plants, was detected in seeds but not in leaves, roots and flowers of the transgenic plants. This expression was accompanied by a significant increase in the levels of free threonine and methionine in the seed. The level of these amino acids also correlated positively with the levels of the bacterial enzyme. No alteration in plant phenotype and ‘average seed weight’ was observed in any of the transgenic plants, indicating that plant growth and seed development were normal. This study demonstrates, for the first time, that the threonine and methionine biosynthetic pathways are active in plant seeds. Thus, targeting of the production of favorable biosynthetic enzymes to plant seeds may represent a desirable molecular approach for production of crop plants with a more balanced nutritional quality.
Purification and biochemical characterization of proteins which bind to the H‐box cis‐elementimplicated in transcriptional activation of plant defense genes
Authors: Yu Lloyd M. , Lamb Christopher J. , Dixon Richard A.
Published: February 2005

The H‐box (CCTACC(N)7CT(N)4A), which occurs three times within the ‐154 to ‐42 region of the bean chal‐cone synthase chs15 promoter, is important for developmental regulation of chs15, and induction of chs15 and coordinately regulated defense genes by elicitors and other stress stimuli. Two protein factors, KAP‐1 and KAP‐2, which recognize conserved features in the H‐box motif, were purified from bean cell suspension cultures by a combination of ion exchange chromatography and DNA affinity chromatography. KAP‐1 is a 97 kDa polypeptide, whereas KAP‐2 comprises two polypeptides of 76 and 56 kDa. KAP‐1 and KAP‐2 also differ in the sensitivity of their DNA‐bound forms to trypsin. Dephosphorylation of KAP‐1 or KAP‐2 affects the mobility of the protein/H‐box binding complex in gel shift assays but does not inhibit DNA binding. Elicitation of bean cell suspensions with glutathione does not affect the total cellular activities of KAP‐1 or KAP‐2, but causes a rapid increase in the specific activities of both factors in the nuclear fraction, consistent with a role for these factors in the signal pathway for elicitor induction of chs15 and related defense genes.
Transcription of two members of a gene family encoding phenylalanine ammonia‐lyase leads to remarkably different cell specificities and induction patterns
Authors: Shufflebottom Diane , Edwards Keith , Schuch Wolfgang , Bevan Michael
Published: February 2005

Phenylalanine ammonia‐lyase (PAL) catalyses the first committed step in the biosynthesis of phenyl‐propanoids, which perform a variety of functions in plant development and in their interactions with the environment. French bean contains a small family of genes encoding PAL and two of these genes, PAL2 and PAL3, have been shown to be differentially expressed at the mRNA level in bean tissues. The transcriptional activities of the PAL2 and PAL3 genes have been investigated by fusing their promoters to the reporter gene β‐glucuronidase (GUS) and transforming these constructs into Arabidopsis, potato and tobacco. The PAL2‐ and PAL3‐GUS constructs exhibited different spatial and temporal patterns of expression during development and in response to environmental stimuli. The consistency of these data with previous mRNA analysis in bean suggests that the differential expression of these two PAL genes is, at least in part, a function of their promoter activities. New patterns of PAL2 and PAL3 promoter activities were also characterized. Some species‐specific differences in GUS expression were observed and these may reflect differences in phenylpropanoid metabolism or the signals that modulate PAL gene transcription.
Altered response to viral infection by tobacco plants perturbed in ubiquitin system.
Authors: Becker Frank , Buschfeld Ellen , Schell Jeff , Bachmair Andreas
Published: February 2005

Tobacco plants can be perturbed in their ubiquitin system by expression of a ubiquitin variant with a lys to arg change at position 48. Plants expressing this ubiquitin variant have a tendency to form necrotic lesions resembling those occurring as defense reactions against some pathogens. Using the interaction between tobacco and tobacco mosaic virus (TMV), it was investigated whether the ubiquitin variant influences plant responses to pathogen infection. Expression of the ubiquitin variant leads to changes in the response of both resistant and susceptible tobacco varieties. Resistant plants form fewer, but darker lesions. In sensitive plants, TMV replication is inhibited, but not completely abolished. Moreover, susceptible plants expressing the ubiquitin variant induce PR‐1 mRNA in response to virus, whereas controls do not express PR‐1 under these conditions.
IgE‐sensitization to predatory mites and respiratory symptoms in Swedish greenhouse workers
Authors: Kronqvist M. , Johansson E. , Kolmodin‐Hedman B. , Öman H. , Svartengren M. , van Hage‐Hamsten M.
Published: February 2005

Background:  Predatory mites are used as biological pesticides worldwide for control of spider mites and other pests in greenhouses. The aim of this study was to evaluate the impact of occupational exposure to Phytoseiulus persimilis and Hypoaspis miles on IgE sensitization among a large group of Swedish greenhouse workers and to examine the relationship between exposure and allergic asthma and rhinoconjunctivitis.

Methods:  A total of 96 greenhouse workers from the southern part of Sweden, who were using the predatory mites for control of pests, were investigated with a questionnaire and a medical examination including lung function test. Blood samples were taken to test for allergen‐specific IgE antibodies to Phytoseiulus persimilis and Hypoaspis miles as well as to Tetranychus urticae, Dermatophagoides pteronyssinus/farinae and Tyrophagus putrescentiae.

Results:  Seventeen of the 96 workers were positive in ImmunoCAP to predatory mites: 17 to P. persimilis (17.7%) and 14 to H. miles (14.6%). Subjects sensitized to predatory mites were significantly more often atopic (13/17), defined as a positive Phadiatop, than those who lacked IgE against these mite species (17/79) (P < 0.01). IgE antibodies to the red spider mite T. urticae were present among 23 subjects. Thirty‐five of the investigated subjects displayed a positive ImmunoCAP to at least one of the investigated mite species. Furthermore, sensitization to any of the mites tested was significantly associated with asthma (OR = 9.3) and rhinoconjunctivitis (OR = 4.3).

Conclusions:  IgE sensitization to predatory mites, P. persimilis and H. miles, is common among greenhouse workers. The findings stress the importance of improved allergen avoidance in greenhouse environments.
Acid–base equilibrium in exhaled breath condensate of allergic asthmatic children
Authors: Carraro S. , Folesani G. , Corradi M. , Zanconato S. , Gaston B. , Baraldi E.
Published: February 2005

Background:  The dysregulation of airway pH control may have a role in asthma pathophysiology. The measurement of exhaled breath condensate (EBC) pH and ammonia levels may be used as a noninvasive method to study acid–base status in the airway of asthmatics.

Methods:  Exhaled breath condensate from 29 allergic stable asthmatic children and 13 healthy controls was collected by cooling exhaled air during tidal breathing. Ammonia was measured by high‐performance liquid chromatography with fluorescence detection. pH was measured after deaeration of EBC samples by bubbling with argon. The children also underwent FENO measurement.

Results:  Both pH and ammonia values in EBC were significantly lower in the asthmatics than in the control group [pH: ICS‐treated (median and interquartile range) 7.70 (7.62–7.74), steroid‐naïve 7.53 (7.41–7.68), controls 7.85 (7.80–7.90), P < 0.01 and P < 0.001, respectively; ammonia: ICS‐treated 476.17 μM (282.50–594.80), steroid‐naïve 253.24 μM (173.43–416.08), controls 788.30 μM (587.29–1310.39), P < 0.05 and P < 0.001, respectively]. Both pH and ammonia values were higher in ICS‐treated than in steroid‐naïve asthmatic children. There was a significant correlation between EBC pH and ammonia concentrations.

Conclusions:  These data show that EBC pH values of stable asthmatic children are lower compared with those of healthy controls and positively correlated with ammonia concentrations, supporting the hypothesis that airway acidification may have a role in the pathobiology of allergic asthma.
Number of offspring and maternal allergy
Authors: Forastiere F. , Sunyer J. , Farchi S. , Corbo G. , Pistelli R. , Baldacci S. , Simoni M. , Agabiti N. , Perucci C. A. , Viegi G.
Published: February 2005

The consistent association seen between family size and childhood allergy has led to the ‘hygiene hypothesis’, namely that a lower frequency of infections in early childhood is associated with an increased risk of asthma and hay fever. Maternal atopy, however, is a strong predictor of childhood asthma and hay fever. If maternal atopy is inversely related to the number of siblings then the role of siblings in the development of childhood atopy, the basic tenet of the ‘hygiene hypothesis’, is challenged. We evaluated the association between number of pregnancies and number of live births with lifetime occurrence of maternal wheeze, asthma, allergic rhinitis, and allergic conjunctivitis in a cross‐sectional study in four areas in Italy. A total of 1755 (35–74 year old) nonsmoking women filled a questionnaire on reproductive history as well as on lifetime occurrence of symptoms/diseases. The number of live births was inversely related to lifetime allergic rhinitis (P‐value for trend = 0.031) and allergic conjunctivitis (P‐value for trend = 0.011). The odds ratios for those with 4+ children (in comparison with those having 0–1) were: 0.53 (95% CI: 0.27–1.04) and 0.42 (95% CI: 0.22–0.81), respectively. A similar trend was seen for number of pregnancies, although not statistically significant. No association was found between number of pregnancies and number of live births with wheeze or asthma. The results may be interpreted as an indication that maternal atopy influences pregnancy outcomes or that pregnancy itself has an effect on maternal atopy.
Bone scintigraphy appearances of incidentally diagnosed soft tissue disorders in musculoskeletal imaging
Authors: Dayanandan A , Ho YY
Published: February 2005

Bone scintigraphy in clinical practice is primarily used for the evaluation of bony pathology. Incidental uptake in soft tissue is likely to be overlooked because it is generally believed to lack specificity. Although bone scintigraphy is not used as a first line investigation for musculoskeletal injuries, abnormal soft tissue uptake should prompt clinicians to reassess patient symptoms. The onus falls on the radiologist and/or nuclear physician to comment on the findings. Several interesting cases are presented here with findings that have been confirmed by other imaging modalities.
Diagnosis of pulmonary embolism: Ventilation perfusion scintigraphy versus helical computed tomography pulmonary angiography
Authors: Macdonald WBG , Patrikeos AP , Thompson RI , Adler BD , Van Der Schaaf AA
Published: February 2005

The present study compared the accuracy of ventilation perfusion scintigraphy (VQS) and CT pulmonary angiography (CTPA) for the diagnosis of pulmonary embolism. This was a prospective observational study of 112 patients with suspected pulmonary embolism (PE) who could be studied with both investigations within 24 h. Results were compared to final diagnosis at completion of 6‐month follow up, using receiver operating characteristic (ROC) analysis. Pulmonary embolism was diagnosed in 27 referred patients (24%). The sensitivity and specificity of VQS and CTPA were similar to that reported from the literature. A normal VQ scan had the highest negative predictive value (100%), while a high‐probability VQ scan had the highest positive predictive value (92%). There was no overall difference (area under the ROC curve (AUC)) between VQS (AUC (95% CI) = 0.82 (0.75,0.89)) and CTPA (AUC = 0.88 (0.81,0.94)) for the diagnosis of PE. Among patients with abnormal chest X‐rays, CTPA (AUC 0.90 (0.83,0.97)) appeared somewhat better than VQS (AUC 0.78 (0.68,0.88)) but this difference did not reach statistical significance. In this instance, CTPA is at least as accurate as VQS and may provide an opportunity to make alternative diagnoses.
Changes in practice of breast cancer radiotherapy 1998–2002: An Australasian survey
Authors: Drummond R , Power A , Evans A , Luxford K , Blakey D , Delaney G , Rodger A
Published: February 2005

In order to assess the impact on radiation oncology practice of the publication of evidence‐based guidelines for technical aspects of therapeutic radiation for breast cancer, the Radiation Oncology Expert Advisory Group of the National Breast Cancer Centre conducted two postal surveys of radiation oncologists practising in Australia and New Zealand. Results from a survey conducted in 1998, prior to distribution of the guidelines, have been published previously. This article reports on results from a survey undertaken in 2002 and contains data from 102 respondents who manage women with breast cancer. The results show several important changes in practice since 1998, including increased use of CT scanning in breast cancer treatment planning and increased use of immobilization devices for patient treatment. There is also evidence of increased attention to technical aspects of treatment planning that reduce the potential risk of treatment toxicity. The influence of the guidelines, the wider availability of modern equipment and results from landmark clinical trials on change in radiation therapy practice is discussed.
Magnetic resonance imaging findings of a foregut duplication cyst of the floor of the mouth in a fetus
Authors: Puvaneswary M , Cassey J
Published: February 2005

The MRI findings are described in a case of foregut duplication cyst arising from the floor of the mouth in a fetus.
Mesenteric panniculitis versus pancreatitis: A computed tomography diagnostic dilemma
Authors: McMenamin DS , Bhuta SS
Published: February 2005

A case of mesenteric panniculitis is described in a patient with CT features mimicking pancreatitis. Emphasis is on the important CT features for accurate diagnosis.
In Defense of the Body: The Effect of Mortality Salience on Female Body Objectification
Authors: Grabe Shelly , Routledge Clay , Cook Alison , Andersen Christie , Arndt Jamie
Published: February 2005

Previous research has illustrated the negative psychological consequences of female body objectification. The present study explores how female body objectification may serve as a defense against unconscious existential fears. Drawing from terror management theory, an experiment was designed to test the potential functionality of female body objectification. Men and women were primed to think about either their own mortality or an aversive control topic, and levels of body objectification were then assessed for both self- and other (women)-objectification. Findings supported the hypothesis that priming mortality would increase both self- and other-objectification among women, and self-objectification among those who derive self-esteem from their body. Implications for this research are discussed.
How Investment in Gender Ideals Affects Well-Being: The Role of External Contingencies of Self-Worth
Authors: Sanchez Diana T. , Crocker Jennifer
Published: February 2005

The present study examined the relationship between investment in gender ideals and well-being and the role of external contingencies of self-worth in a longitudinal survey of 677 college freshmen. We propose a model of how investment in gender ideals affects external contingencies and the consequences for self-esteem, depression, and symptoms of disordered eating. Specifically, we find that the negative relationship between investment in gender ideals and well-being is mediated through externally contingent self-worth. The model showed a good fit for the overall sample. Comparative model testing revealed a good fit for men and women as well as White Americans, Asian Americans, and African Americans.
14 Tuberculosis Skin Test in Chronic Hemodialysis Patients
Authors: Urbančič A , Premru V , Knap B , Benedik M , Ponikvar‐Buturović J , Ponikvar R , Bren A F
Published: February 2005

The incidence of tuberculosis (TB) is increasing worldwide due to increasing number of immigrants during recent years. This increases the risk of TB in high risk groups of patients. Cellular immunity has been demonstrated to be impaired in chronic hemodialysis (CHD) patients. Accordingly the incidence of TB in dialysis patients was presumably higher than in the general population. In an attempt to detect patients at risk for TB among CHD patients a crossover evaluation of tuberculosis skin test was performed in our dialysis center. Some preliminary results are presented here. Among 203 tested CHD patients 139 (68.5%) did not respond. In 64 (31.5%) patients, infiltrate was detected. The diameter of the infiltrate was 1 to < 5 mm in 4/64 patients (6.25%); it was 5 to < 10 mm in 18/64 pts (28.1%) and > 10 mm in 36/64 patients (56.3%). Mean age of tested patients on CHD for more than a year was similar in infiltrate positive subgroups of patients: 59.7, 58.7, and 55.9 years respectively. There are questions whether younger CHD patients have a better preserved cellular immunity; whether markedly positive skin test reaction for TB is connected with triggered cellular immune response to TB or is it a sign of subclinical TB disease. Chronic hemodialysis patients with highly positive skin TB test should be actively examined for TB disease.
16 Treating of Nasal Carriers with Mupirocin and Prevention of Staphylococcus aureus Peritonitis and Esi/Ti in Peritoneal Dialysis Patients
Authors: Guček A , Karas B , Pajek J , Benedik M
Published: February 2005

Several studies found an increased risk of Staphylococcus aureus (SA) exit‐site and/or tunnel infections (ESI/TI) or peritonitis (P) in SA nasal carriers. The aim of our study was to determine efficacy of local preventive therapy with mupirocin in nasal carriers of SA. In this prospective study, from January 1997 to December 2003, 127 patients (pts) were included and observed for 4696 pt‐months. The ones with one or more positive smears were treated with mupirocin for five consecutive days, twice daily, every month. In acute or chronic ESI/TI with SA, the mupirocin was applied during daily exit‐site care. The pts with good or perfect exit‐sites were not treated. For statistic analysis, the proportion test and Pearson¢s correlation test were used. Among our pts the number of nasal carriers has decreased significantly (P < 0.05) from 1997 (49.3%) to 2003 (18.0%). Simultaneously, the rates of SA peritonitis dropped from 0.032 episode/pt year in 1997 to 0.022 in 2003 (P = 0.25). Significantly less (P = 0.005) new SA ESI/TI were registered, from 0.113 episode/pt‐year in 1997 to 0.018 in 2001 and 0.022 in 2003. Two cases of mupirocin‐resistant SA were isolated, both in 2003, present in nasal as well as in ESI/TI swab. Diagnosis and treatment of SA nasal carriers among PD pts has proved to be important. The decrease in incidence of new ESI/TI was statistically significant, while peritonitis rates decreased less significantly. Mupirocin‐resistance in our study is low. When peritoneal catheter exit‐site is good or perfect, treatment with mupirocin is not necessary.