bluuebunny/crossref_metadata_2025_split huggingface dataset:
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

SummaryThe 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

SummaryA 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

SummaryTo 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

SummaryThe 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

SummaryIn 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

SummaryThe 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

SummaryPhenylalanine 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

SummaryTobacco 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.
Bone scintigraphy appearances of incidentally diagnosed soft tissue disorders in musculoskeletal imaging
Authors: Dayanandan A , Ho YY 
Published: February 2005

SummaryBone 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

SummaryThe 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

SummaryIn 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.
24 Response on Epoetin Treatment is Poor in Hemodialysed Patients with Low Creatinine and Albumin Values
Authors: Knap B , Lainščak M , Varl J , Bren AF 
Published: February 2005

Background Anemia frequently accompanies chronic renal failure in patients treated with hemodialysis (HD). Despite treatment with erythropoetin up to 10% of patients still present with pronounced anemia. In the present study we wanted to identify simple laboratory parameters or clinical characteristics, associated to anemia in patients treated with erythropoetin.Methods During May 2003 we screened all patients at our hemodialysis center for the presence of severe anemia (Hemoglobin (Hb) 100 or lower, Group 1) and compared them to the patients without anemia (Hb 120 or higher, Group 2). Patients with chronic bleeding or the ones receiving transfusion were excluded. Basic demographic characteristics, blood pressure, and several biochemical markers were obtained for the enrolled patients. All data were tested for significance using Student's t‐test.Results Out of 177 patients, 34 (Group 1) and 38 (Group 2) met the inclusion criteria. There was no difference in age, gender, time on HD and blood pressure (148 ± 22/81 ± 13 mm Hg vs. 151 ± 33/81 ± 15 mm Hg). Next to significant difference all hematological parameters (Hb 92 ± 8 vs. 127 ± 6; Ht 0.29 ± 0.03 vs. 0.40 ± 0.02; Ery 3.04 ± 0.32 vs. 4.17 ± 0.32; P < 0.001 for all) patients with anemia received a higher weekly dose of erythropoetin (12000 ± 5205 IE vs. 7825 ± 4829 IE, P < 0.001), had lower albumin (35.6 ± 3.4 vs. 38.1 ± 2.8, P < 0.001) and creatinine concentration (798 ± 218 vs. 932 ± 266, P < 0.005). For other markers such as ferritin, iPTH, folic acid, and vitamin B12 we observed a trend to higher values in anemic patients.Conclusions Our data suggest that malnutrition and low muscle mass are associated to poor response of treatment with erythropoetin in patients with chronic renal failure treated with HD. Larger sample and prospective follow up could identify further simple markers of inadequate response to the treatment of anemia.
27 Citrate Anticoagulation for Single‐Needle Hemodialysis: Safety and Efficacy
Authors: Buturović‐Ponikvar J , Gubenšek J , Ponikvar R 
Published: February 2005

Background  Single‐needle (SN) hemodialysis can be the only option for selected patients with difficulties with vascular access. Full heparinization is required to avoid clotting in the circuit. In patients at risk of bleeding, citrate could be the alternative anticoagulant. The aim of our retrospective clinical study was to evaluate safety and efficacy of regional citrate anticoagulation (RCA) during SN hemodialysis.Patients and methods  Regional citrate anticoagulation was performed using 4% trisodium citrate, 1 m CaCl2 and dialysate with 0 mmol/L Ca, 0.5 mmol/L Mg and 0 g/L glucose, Na was set at 138 mmol/L and HCO3 at 28 mmol/L. Safety was assessed by percent of procedures that were terminated prematurely or changed to another modality due to RCA‐related complications and by incidence of important hypocalcemia, defined as decrease of iCa from start of dialysis > 0.2 mmol/L or decrease > 0.05 mmol/L to a value of < 0.8 mmol/L. Efficacy was evaluated by visual assessing of clot formation in arterial and venous bubble traps and dialyzer, after completing dialysis, by scoring: grade 5 (<10 fibers clotted) to 0 (>20% fibers clotted). It was considered insufficient if the grade was ≤ 3 or a significant clot in bubble traps occurred.Results  In retrospective analysis of 41 protocols, important hypocalcemia was recorded in 34% of cases. 5% of procedures were terminated prematurely. Median dialyzer grade was 5 (interquartile range 4–5, N = 36). Anticoagulation was insufficient in 17% (6/36) of procedures, in these cases neither citrate infusion rate was lower nor coagulation time shorter. None of the systems clotted. Thirty protocols were filled in completely; average parameters are shown in the table below. In the first hour iCa decreased in 67% of procedures for 0.08 ± 0.05 mmol/L, those procedures had significantly lower starting calcium rate (6.4 ± 0.9 vs. 7.3 ± 1.1 mmol/L, P = 0.02) and higher starting iCa (1.02 ± 0.13 vs. 0.89 ± 0.14 mmol/L, P = 0.02). Over the entire procedure iCa decreased in 80% of cases for 0.17 ± 0.09 mmol/L, there was significant but small increase in Na (135 ± 4 vs. 137 ± 4 mmol/L, P ≤ 0.01) and no increase in HCO3 at the end of hemodialysis (Table A27).

Parameter
Value

blood flow (mL/min)
244 ± 27

starting rate of 4% Na citrate (mL/h)
191 ± 19

citrate rate (percent of mean blood flow)
2.6% ± 0.4%

starting rate of 1 m CaCl2 (mL/h)
6.7 ± 1.1

Conclusions  Citrate anticoagulation during single‐needle hemodialysis is a safe and effective procedure in the majority of patients. CaCl2 infusion rate should often be increased to correct hypocalcemia. Close monitoring of iCa is mandatory. Increased starting dose of CaCl2 should be evaluated.