bluuebunny/crossref_metadata_2025_split huggingface dataset:
Leghaemoglobin gene transcription is triggered in a single cell layer in the indeterminate nitrogen‐fixing root nodule of alfalfa
Authors: de Billy Françoise , Barker David George , Gallusci Philippe , Truchet Georges 
Published: February 2005

SummaryThe pattern of leghaemoglobin (Lb) gene expression within the indeterminate nitrogen‐fixing root nodule of Medicago species such as alfalfa has been examined by means of ‘in‐situ’ hybridization. Hybridization of alfalfa nodule sections with radiolabelled Lb RNA probes clearly shows that Lb gene expression is triggered in a single cell layer just prior to the start of the nitrogen fixation zone. This cell layer is part of a unique region (interzone II‐III) in which invaded cells are rich in amyloplasts. Lb transcripts can only be detected in cells which have been invaded by the microsymbiont Rhizobium. The conclusion that Lb gene activation precedes the start of nitrogen fixation is further supported by the observation that Lb transcripts are already present in immature non‐nitrogen‐fixing nodules of alfalfa. The addition of 5 mM KNO3 to the growth medium of nodulated M. truncatula (a diploid autogamous species of Medicago) results in a significant drop in both nitrogen‐fixing activity and nodular Lb mRNA levels. Short periods of exposure to nitrate (24 h) lead to the disappearance of Lb transcripts from the proximal region of the hybridizing zone, whilst longer exposures (72 h) lead to a marked decrease in the intensity of the hybridization signal within the amyloplast‐rich interzonal region.
An improved procedure for the isolation of plasmodesmata embedded in clean maize cell walls
Authors: Kotlizky Guy , Shurtz Shomrat , Yahalom Avital , Malik Zvi , Traub Otto , Epel Bernard L. 
Published: February 2005

SummaryA cell wall preparation of high purity was obtained using a procedure which involved repeated grindings of etiolated maize mesocotyl tissue and filtration through 200 mesh nylon cloth, followed by cell disruption via a nitrogen disruption bomb, and recovery of the cell walls via filtration. The cell wall fraction was free of particulate contaminants as determined both by phase‐contrast and electron microscopy. The only membrane components found associated with the wall fraction as determined by electron microscopy were pladmodesmata embedded in the cell wall. The specific concentration of PAP26, a plasmodesmatal‐associated polypeptide, was greatly increased in the cleanest cell wall fraction. A second plasmodesmatal‐associated protein, PAP27, which was previously shown to be associated with the neck region of the plasmodesmata, was diminished as a result of passage through the nitrogen disruption bomb suggesting a partial fragmentation of the plasmodesmata. In addition to PAP26, the specific concentrations of at least three other cell wall‐associated polypeptides with molecular weights of 80, 21 and 18 kDa, as revealed by SDS‐PAGE, were also increased greatly in the cleanest cell wall fraction.
Indole‐3‐acetic acid homeostasis in transgenic tobacco plants expressing the Agrobacterium rhizogenes rolB gene
Authors: Nilsson Ove , Crozier Alan , Schmülling Thomas , Sandberg Göran , Olsson Olof 
Published: February 2005

SummaryThe rolB gene of the plant pathogen Agrobacterium rhizogenes has an important role in the establishment of hairy root disease in infected plant tissues. When expressed as a single gene in transgenic plants the RolB protein gives rise to effects indicative of increased auxin activity. It has been reported that the RolB product is a β‐glucosidase and proposed that the physiological and developmental alterations in transgenic plants expressing the rolB gene are the result of this enzyme hydrolysing bound auxins, in particular (indole‐3‐acetyl)‐β‐D‐glucoside (IAGluc), and thereby bringing about an increase in the intracellular concentration of indole‐3‐acetic acid (IAA). Using tobacco plants as a test system, this proposal has been investigated in detail. Comparisons have been made between the RolB phenotype and that of IaaM/iaaH transformed plants overproducing IAA. In addition, the levels of IAA and IAA amide and IAA ester conjugates were determined in wild‐type and transgenic 35S‐rolB tobacco plants and metabolic studies were carried out with [13C6]IAA [2′‐14C]IAA, [14C]IAGluc, [5‐3H]‐2‐o‐(indole‐3‐acetyl)‐myo‐inositol and [14C]indole‐3‐acetylaspartic acid. The data obtained demonstrate that expression of the rolB encoded protein in transgenic tobacco does not produce a phenotype that resembles that of IAA over producing plants, does not alter the size of the free IAA pool, has no significant effect on the rate of IAA metabolism, and, by implication, appears not to influence the overall rate of IAA biosynthesis. Furthermore, the in vivo hydrolysis of IAGluc, and that of the other IAA conjugates that were tested, is not affected. On the basis of these findings, it is concluded that the RolB phenotype is not the consequence of an increase in the size of the free IAA pool mediated by an enhanced rate of hydrolysis of IAA conjugates.
Action of a pure xyloglucan endo‐transglycosylase (formerly called xyloglucan‐specific endo‐(1‐4)‐β‐d‐glucanase) from the cotyledons of germinated nasturtium seeds
Authors: Fanutti Cristina , Gidley Michael J. , Reid J.S. Grant 
Published: February 2005

SummaryThe action on tamarind seed xyloglucan of the pure, xyloglucan‐specific endo‐(1→4)‐β‐D‐glucanase from nasturtium (Tropaeolum majus L.) cotyledons has been compared with that of a pure endo‐(1→4)‐β‐D‐glucanase (‘cellulase’) of fungal origin. The fungal enzyme hydrolysed the polysaccharide almost completely to a mixture of the four xyloglucan oligosaccharides:image
Exhaustive digestion with the nasturtium enzyme gave the same four oligosaccharides plus large amounts of higher oligosaccharides and higher‐polymeric material. Five of the product oligosaccharides (D,E,F,G,H) were purified and shown to be dimers of oligosaccharides A to C. D (glc8xyl6) had the structure A→A, H (glc8xyl6gal4) was C→C, whereas E (glc8xyl6gal), F (glc8xyl6gal2) and G (glc8xyl6gal3) were mixtures of structural isomers with the appropriate composition. For example, F contained B2→B2 (30%), A→C (30%), C→A (20%), B2B1 (15%) and others (about 5%). At moderate concentration (about 3 mM) oligosaccharides D to H were not further hydrolysed by the nasturtium enzyme, but underwent transglycosylation to give oligosaccharides from the group A, B, C, plus higher oligomeric structures. At lower substrate concentrations, hydrolysis was observed. Similarly, tamarind seed xyloglucan was hydrolysed to a greater extent at lower concentrations. It is concluded that the xyloglucan‐specific nasturtium‐seed endo‐(1→4)‐β‐D‐glucanase has a powerful xyloglucan‐xyloglucan endo‐transglycosylase activity in addition to its known xyloglucan‐specific hydrolytic action. It would be more appropriately classified as a xyloglucan endo‐transglycosylase. The action and specificity of the nasturtium enzyme are discussed in the context of xyloglucan metabolism in the cell walls of seeds and in other plant tissues.
Light‐regulated expression of the Arabidopsis thaliana ferredoxin A gene involves both transcriptional and post‐transcriptional processes
Authors: Vorst Oscar , van Dam Frans , Weisbeek Peter , Smeekens Sjef 
Published: February 2005

SummaryFerredoxin is part of the photosynthetic apparatus of the chloroplast and is encoded in the nucleus. In Arabidopsis thaliana expression of the ferredoxin A gene is influenced by both the presence of chloroplasts and light. Tobacco plants transformed with a ferredoxin promoter–GUS fusion gene showed a tissue‐specific and light‐dependent expression pattern. The effect of light on the expression of the fusion gene in transgenic seedlings was only two‐ to fourfold, which is less pronounced than the 20‐fold effect in Arabidopsis itself. Run‐on transcription assays with nuclei isolated from Arabidopsis revealed a twofold modulation of transcriptional activity of the ferredoxin A gene under the influence of light. These results suggest the involvement of post‐transcriptional processes in light‐regulated gene expression.A ferredoxin promoter deletion series ranging from ‐1205 to ‐143 was studied. All but the smallest deletion construct (at position ‐143 relative to the translation start site) showed comparable expression levels in mature leaves, suggesting the presence of a positive regulating element between ‐269 and ‐143. The same pattern of tissue specificity was found in all promoter deletions studied. Expression of the fusion genes is high in all chloroplast‐containing cells: mesophyll, chlorenchyma, paravascular tissue, epidermal and stomatal guard cells and trichomes. Transgenic seedlings treated with norflurazon, which blocks the development of green chloroplasts, showed a two‐ to fourfold reduction in GUS expression for all constructs. In Arabidopsis seedlings the effect of norflurazon on the expression of the ferredoxin A was eightfold. This again can be explained by the need for post‐transcriptional processes of the regulated gene expression of Arabidopsis ferredoxin A.
Developmentally regulated expression of the wound‐ and pathogen‐responsive tomato anionic peroxidase in green fruits
Authors: Sherf Bruce A. , Kolattukudy P.E. 
Published: February 2005

SummaryA highly anionic peroxidase (TAP) isolated from tomato was thought to be induced only upon wounding or fungal attack as this protein and its mRNA could not be detected in the vegetative tissues of healthy tomato but accumulated to very high levels in tissues responding to wounding or fungal attack. We now demonstrate a pattern of developmentally regulated expression of the tap gene in the exocarp of green, but not red, tomato fruits. Detaching green fruits from the vine, however, leads to the complete loss of the tap transcript within 72 h. Administration of various growth regulators through the pedicel of detached green fruits revealed that cytokinin was able to sustain a level of tap transcripts in 72 h detached fruits comparable with that found in fruits attached to the vine. Wound‐induced expression of tap was confined to the healing periderm of green fruits but was not detectable in the wound‐tissue of post‐climacteric ripening fruits. These results suggest that the developmental and wound regulated expression of tap may occur by separate or overlapping signal transduction pathways. Treatment of tomato plants and green fruits with 2‐chloro‐ethylphosphonic acid (ethephon) demonstrated that ethylene does not effect the developmental suppression of tap in post‐climacteric fruits, nor does it alter the normal levels of tap transcripts in either healthy or wound‐healing tomato tissues.
Localization of vicilin genes via polymerase chain reaction on microisolated field bean chromosomes
Authors: Macas Jiri , Weschke Winfriede , Bäumlein Helmut , Pich Uta , Houben Andreas , Wobus Ulrich , Schubert Ingo 
Published: February 2005

SummaryA new technique is reported for the physical mapping of low copy DNA sequences on plant chromosomes. Individual chromosomes were microisolated and their DNA used as the target for the polymerase chain reaction in order to identify the chromosome carrying a specific gene sequence. The use of defined translocation chromosomes further refined the resolution of the method to a subchromosomal level. To demonstrate the applicability of the procedure genes have been localized coding for vicilin seed storage proteins on the field bean Vicia faba L. in a region which includes the centromere and the proximal parts of the short and the long arms of chromosome II.
Probiotics in the treatment of atopic eczema/dermatitis syndrome in infants: a double‐blind placebo‐controlled trial
Authors: Viljanen M. , Savilahti E. , Haahtela T. , Juntunen‐Backman K. , Korpela R. , Poussa T. , Tuure T. , Kuitunen M. 
Published: February 2005

Background:  Probiotic bacteria are suggested to reduce symptoms of the atopic eczema/dermatitis syndrome (AEDS) in food‐allergic infants. We aimed to investigate whether probiotic bacteria have any beneficial effect on AEDS.Methods:  Follow‐up of severity of AEDS by the Severity Scoring of Atopic Dermatitis (SCORAD) index in 230 infants with suspected cow's milk allergy (CMA) receiving, in a randomized double‐blinded manner, concomitant with elimination diet and skin treatment, Lactobacillus GG (LGG), a mixture of four probiotic strains, or placebo for 4 weeks. Four weeks after the treatment, CMA was diagnosed with a double‐blind placebo‐controlled (DBPC) milk challenge in 120 infants.Results:  In the whole group, mean SCORAD (at baseline 32.5) decreased by 65%, but with no differences between treatment groups immediately or 4 weeks after the treatment. No treatment differences were observed in infants with CMA either. In IgE‐sensitized infants, however, the LGG group showed a greater reduction in SCORAD than did the placebo group, −26.1 vs−19.8 (P = 0.036), from baseline to 4 weeks after the treatment. Exclusion of infants who had received antibiotics during the study reinforced the findings in the IgE‐sensitized subgroup.Conclusion:  Treatment with LGG may alleviate AEDS symptoms in IgE‐sensitized infants but not in non‐IgE‐sensitized infants.
IgE antibodies to ω‐5 gliadin in children with wheat‐induced anaphylaxis
Authors: Daengsuwan T. , Palosuo K. , Phankingthongkum S. , Visitsunthorn N. , Jirapongsananuruk O. , Alenius H. , Vichyanond P. , Reunala T. 
Published: February 2005

Background:  Wheat can cause severe immunoglobulin E (IgE)‐mediated systemic reactions including anaphylaxis but knowledge on relevant wheat allergens at the molecular level is scanty.Methods:  Seven children (aged from 6 months to 13 years) experiencing from 2 to 10 anaphylactic reactions in a year after eating food‐containing wheat were examined. Purified ω‐5 gliadin was used as an allergen in IgE enzyme‐linked immunosorbent assay (ELISA) and in skin prick testing (SPT). Wheat CAP radioallergosorbent test (RAST) and SPT were also examined.Results:  All seven anaphylactic children, but none of 15 control subjects had IgE antibodies to ω‐5 gliadin in ELISA. Five of the six tested anaphylactic children showed positive SPT to ω‐5 and crude gliadin, and all seven had positive wheat CAP RAST and SPT. One child was challenged with wheat, which caused anaphylaxis. After adherence to a wheat‐free diet four children remained symptomless and three experienced one to two anaphylactic reactions.Conclusion:  The present results show that wheat ω‐5 gliadin is a major sensitizing allergen in children with wheat‐induced anaphylaxis. They also suggest that ω‐5 gliadin IgE ELISA could be used as a diagnostic test for this severe allergy.
Improvement of chronic idiopathic urticaria with l‐thyroxine: a new TSH role in immune response?
Authors: Aversano M. , Caiazzo P. , Iorio G. , Ponticiello L. , Laganá B. , Leccese F. 
Published: February 2005

Background:  The association between chronic idiopathic urticaria (CIU) and autoimmune thyroiditis (AT) is known, as well as major prevalence of antithyroid antibodies in the allergical subjects and other autoimmune diseases. We have evaluated the effects of l‐thyroxine on clinical symptoms of CIU in AT patients suggesting the hypothesis of a new thyroid‐stimulating hormone (TSH) role in immune system.Methods:  In 20 female patients with CIU + AT, both hypothyroid and euthyroid, we have investigated the therapeutic effects of l‐thyroxine dosed to suppress the TSH. Free‐T3, Free‐T4, TSH, antithyroperoxidase and antithyroglobulin antibodies, total immunoglobulin (Ig)E, Rheuma test and eritro‐sedimentation rate were monitored during treatment.Results:  In 16 patients a strong decrease of urticaria symptoms has happened after 12 weeks. The TPO Ab and HTG Ab clearly decreased in 14 patients. Furthermore, in two patients with rheumatoid arthritis and in two patients with pollen allergy a strong decrease of rheuma test titer and total IgE has happened.Conclusion:  The reason of AT is associated to CIU and others allergical and autoimmune diseases is poorly known. The exclusive hormonal therapy reduces the symptoms of CIU and inflammatory response in many chronic diseases associated to AT. We suggest a stimulatory effect of TSH able to produce considerable changes of the immune response and immune tolerance in patients with AT causing target organs damage.The causal mechanism involves immune, nervous and endocrine system, sharing a common set of hormones, cytokines and receptors, in a unique totally integrated loop (the neuro–immuno–endocrine axis).
The reduction of rhinitis symptoms by nasal filters during natural exposure to ragweed and grass pollen
Authors: O'Meara T. J. , Sercombe J. K. , Morgan G. , Reddel H. K. , Xuan W. , Tovey E. R. 
Published: February 2005

Background:  Prototype nasal filters were developed to collect inhaled pollen. This study evaluated the efficacy of the filters for prevention of rhinitis symptoms during acute outdoor pollen exposure.Methods:  A randomized double‐blind design was used. Subjects (n = 46) with a history of autumn exacerbation of rhinitis and positive skin test to ragweed, Bermuda and/or Bahia grass wore either active or placebo nasal filters for 2 h in autumn in a park containing these species. Major and Total Symptoms scores were recorded at 0, 30, 60, 90 and 120 min.Results:  Subjects wearing active nasal filters had significantly reduced scores, at all time‐points compared with placebo group (all P < 0.05). Of 14 individual symptoms measured, seven were significantly reduced (number of sneezes, runny nose, itchy nose, sniffles, itchy throat; itchy eyes and watery eyes) and another three showed a trend towards lower severity. The nasal filters also enabled the resolution of existing symptoms. Maximal difference in symptoms was seen immediately after subjects had spent 20 min sitting beside a large patch of ragweed.Conclusion:  This is the first clinical trial of a nasal filter. The results suggest it has potential for enhancing rhinitis management during acute allergen exposure.
Incidental heterotopic pregnancy demonstrated on magnetic resonance imaging
Authors: Tan PL , Ridley LJ 
Published: February 2005

SummaryWe report a case of an assisted pregnancy in an asymptomatic woman who was found to have an extrauterine mass on ultrasound and MRI. This complex mass had equivocal imaging features and was found to be a ruptured ovarian ectopic pregnancy at surgery. This case illustrates that vigilance is required regarding the possibility of coexisting ectopic and intrauterine pregnancy following assisted conception, even in entirely asymptomatic cases.
11 Thrombolysis of Permanent Hemodialysis Catheters
Authors: Premru V 
Published: February 2005

Outline  Occlusion of hemodialysis (HD) catheter and infection are by far the most frequent reasons for removal of the catheter. As an increasing number of patients depend on a catheter as a vascular access for an extended period, maintenance of HD catheter patency is extremely important. One approach in case of catheter occlusion is thrombolysis. We describe our experience with HD catheter thrombolysis in rare patients with extreme access problems.Patients and methods  In four patients (aged 77, 69, 64 and 28 years, all female) hemodialyzed via permanent HD catheters, occlusion occurred and thrombolysis was attempted. Primary patency periods were: 112 months for one surgically implanted subclavian vein silastic catheter, 74 and 46 days for two percutaneously inserted jugular vein catheters and 30 days for one surgically implanted right intra‐atrial catheter. Recombinant human tissue plasminogen activator (TPA) or urokinase was used 13 times each as a thrombolytic agent in recommended doses for catheter lysis. Heparin lock was also used three times.Results  In the young patient with the intra‐atrial catheter, r‐TPA was used 13 times with modest success even in the short‐term: restoration of flow for average 11 (range, 1–23) days. Urokinase was used 13 times in the other three patients: average secondary patency time was 79 (3–109) days. Heparin was effective in restoring patency for 29 (22–34) days. No complications of thrombolytic therapy were noted.Discussion  Thrombolysis of occluded HD catheter is sometimes the only way to extend its functioning time. Especially in difficult‐to‐manage cases with exhausted vascular access sites, even for catheter insertion, this might be of vital importance. In our experience, in rare patients thrombolysis is needed frequently. It can be repeated safely. Unfortunately, the effects of each treatment seem to be only short‐lived. Beside its allergogenic potential and limited effectiveness, this therapy is also very expensive.
12 Genotype and Transmission of Hepatitis C Virus in Hemodialysis Units
Authors: Meši´ E , Tihić N , Resić H 
Published: February 2005

Objectives  Hepatitis C virus (HCV) infection is spread worldwide with significant geographical differences in the prevalence and genotypes. The aim of this study was to analyse HCV infection in hemodialysis (HD) units in Bosnia and Herzegovina (BH), through genotypes and routes of transmission.Design and Methods  We analysed data from Renal Registry of BH for the years 2002 and 2003 in order to estimate prevalence and incidence of anti‐HCV positive patients in 23 HD units in BH (ELISA III). Then we tested 88 anti‐HCV positive patients from Tuzla for HCV RNA using reverse transcription polymerase chain reaction (Amplicor Roche diagnostic commercial test) and genotype with method of reverse hibridization of amplified sample material (Innolipa HCV II commercial test).Results  In 2002 we registered a prevalence of 39.1 and an incidence of 3.1% of anti‐HCV positive HD patients in 23 HD units, and in 2003 a prevalence of 29.3 and an incidence of 2.04% in 24 HD units. Prevalence of anti‐HCV positive patients was significantly different in HD units, from 14.3 to 69.1%. From 88 tested anti‐HCV positive HD patients in Tuzla HD unit, we found 76 HCV RNA positive patients (87.36%). The genotype of HCV was analysed in 59 of these patients and we found, unusual for Europe, genotype 4 in 37 (62.7%), genotype 1b in 17 (28.8%) and 1a in five patients (8.5%). A total of 61% of seroconversions happened in 1999, 2000 and 2001. In 2001, we separated dialysis machines for anti‐HCV positive and anti‐HCV negative patients, and in 2003 we had nine seroconversions (genotype 4 in five patients). These patients had no blood transfusions.Conclusions  Prevalence and incidence of anti‐HCV positive hemodialysis patients is still high in BH and origin of genotype 4 HCV is not clear. Nosocomial spreading of HCV plays a significant and probably the most important role in transmission of HCV in HD units.