bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification and expression analysis of two inorganic C‐ and N‐responsive genes encoding novel and distinct molecular forms of eukaryotic phosphoenolpyruvate carboxylase in the green microalga Chlamydomonas reinhardtii
Authors: Mamedov Tarlan G. , Moellering Eric R. , Chollet Raymond 
Published: June 2005

SummaryPhosphoenolpyruvate carboxylase (PEPC [Ppc]) has been previously purified and characterized in biochemical and immunological terms from two green microalgae, Chlamydomonas reinhardtii and Selenastrum minutum. The findings indicate that these algae possess at least two distinct PEPC enzyme‐forms, homotetrameric Class‐1 and heteromeric Class‐2, that differ significantly from each other and their plant and prokaryotic counterparts. Surprisingly, however, green‐algal PEPC has been unexplored to date in molecular terms. This study reports the molecular cloning of the two Ppc genes in C. reinhardtii (CrPpc1, CrPpc2), each of which is transcribed in vivo and encodes a fully active, recombinant PEPC that lacks the regulatory, N‐terminal seryl‐phosphorylation domain that typifies the vascular‐plant enzyme. These distinct catalytic subunit‐types differ with respect to their (i) predicted molecular mass (∼108.9 [CrPpc1] versus ∼131.2 kDa [CrPpc2]) and critical C‐terminal tetrapeptide; and (ii) immunoreactivity with antisera against the p102 and p130 polypeptides of S. minutum PEPC1/PEPC2 and PEPC2, respectively. Only the Ppc1 transcript encodes the p102 catalytic subunits common to both Class‐1 and Class‐2 enzyme‐forms in C. reinhardtii. The steady‐state transcript levels of both CrPpc1/2 are coordinately up‐/down‐regulated by changes in [CO2] or [NH] during growth, and generally mirror the response of cytoplasmic glutamine synthetase (Gs1) transcript abundance to changes in inorganic [N] at 5% CO2. These collective findings provide key molecular insight into the Ppc genes and corresponding PEPC catalytic subunits in the eukaryotic algae.
Cold‐inducible zinc finger‐containing glycine‐rich RNA‐binding protein contributes to the enhancement of freezing tolerance in Arabidopsis thaliana
Authors: Kim Yeon‐Ok , Kim Jin Sun , Kang Hunseung 
Published: June 2005

SummaryGlycine‐rich RNA‐binding proteins (GR‐RBPs) have been implicated to play roles in post‐transcriptional regulation of gene expression in plants under various stress conditions, but the functional roles of GR‐RBPs under stress conditions remain to be verified. Here, we examine the biological roles of a GR‐RBP, designated atRZ‐1a, in Arabidopsis thaliana under stress conditions. atRZ‐1a was expressed ubiquitously in various Arabidopsis organs including stems, roots, leaves, flowers, and siliques. The transcript level of atRZ‐1a increased markedly by cold stress, whereas its expression was marginally downregulated by drought stress or abscisic acid treatment. Germination and seedling growth of the loss‐of‐function mutants were retarded remarkably compared with those of the wild type under cold stress. In contrast, the transgenic Arabidopsis plants that overexpress atRZ‐1a displayed earlier germination and better seedling growth than the wild type under cold stress. Moreover, the atRZ‐1a‐overexpressing transgenic Arabidopsis plants were more freezing tolerant than the wild‐type plants. Heterologous expression of atRZ‐1a in Escherichia coli demonstrated that the E. coli cells expressing atRZ‐1a displayed much higher growth rate than the non‐transformed cells after cold shock. These results provide evidence that atRZ‐1a affects seed germination and seedling growth under low temperature and plays a role in the enhancement of freezing tolerance in Arabidopsis plants.
The Medicago CDKC;1‐CYCLINT;1 kinase complex phosphorylates the carboxy‐terminal domain of RNA polymerase II and promotes transcription
Authors: Fülöp Katalin , Pettkó‐Szandtner Aladàr , Magyar Zoltán , Miskolczi Pál , Kondorosi Éva , Dudits Dénes , Bakó László 
Published: June 2005

SummaryThe Ms;CDKC;1 kinase is structurally similar to those cyclin‐dependent kinases (CDKs) that are not involved directly in cell cycle regulation. The presence of a PITAIRE motif in Ms;CDKC;1 suggests that it interacts with cyclins different from known PSTAIRE/PPTALRE kinase regulatory subunits. Here we demonstrate that a Medicago CYCLINT (CYCT) protein is a specific interactor of Ms;CDKC;1 and the interaction between these two proteins gives rise to an active kinase complex that localizes to the nucleus and phosphorylates the carboxy‐terminal YSPTSPS heptapeptide repeat domain (CTD) of the largest subunit of RNA polymerase II in vitro. Mutation of Ser to Ala at position 5 within the heptapeptide repeat abolishes substrate phosphorylation by the Ms;CDKC;1 kinase complex. Furthermore, our data show that addition of the Medicago CDKC;1‐CYCT;1 heterodimer completely restored the transcriptional activity of a HeLa nuclear extract depleted of endogeneous CDK9 kinase complexes. Together, these results indicate that the Medicago CDKC;1‐CYCT;1 complex is a positive regulator of transcription in plants and has a role similar to the CDK9/cyclin T complex of human positive transcription elongation factor P‐TEFb.
The plant Mo‐hydroxylases aldehyde oxidase and xanthine dehydrogenase have distinct reactive oxygen species signatures and are induced by drought and abscisic acid
Authors: Yesbergenova Zhazira , Yang Guohua , Oron Einav , Soffer Dana , Fluhr Robert , Sagi Moshe 
Published: June 2005

SummaryThe plant molybdenum‐cofactor (Moco) and flavin‐containing enzymes, xanthine dehydrogenase (XDH; EC 1.2.1.37) and aldehyde oxidase (AO; EC 1.2.3.1) are thought to play important metabolic roles in purine metabolism and hormone biosynthesis, respectively. Their animal counterparts contribute to reactive oxygen species (ROS) production in numerous pathologies and here we examined these enzymes as potential sources of ROS in plants. Novel in‐gel assay techniques and Moco sulfurase mutants, lacking a sulfur ligand in their Moco active center, were employed to demonstrate that the native tomato and Arabidopsis XDHs are capable of producing O, but not H2O2, while the animal counterpart was shown to produce both, O and H2O2. Superoxide production was dependent on Moco sulfuration when using hypoxanthine/xanthine but not NADH as substrates. The activity was inhibited by diphenylene iodonium (DPI), a suicide inhibitor of FAD containing enzymes. Analysis of XDH in an Arabidopsis Atxdh1 T‐DNA insertion mutant and RNA interference lines revealed loss of O activity, providing direct molecular evidence that plant XDH generates superoxides. Contrary to XDH, AO activity produced only H2O2 dissimilar to native animal AO, that can produce O as well. Surprisingly, H2O2 accumulation was not sensitive to DPI. Plant ROS production and transcript levels of AO and XDH were rapidly upregulated by application of abscisic acid and in water‐stressed leaves and roots. These results, supported by in vivo measurement of ROS accumulation, indicate that plant AO and XDH are possible novel sources for ROS increase during water stress.
Seasonal and perennial allergic rhinitis: is this classification adherent to real life?
Authors: Ciprandi G. , Cirillo I. , Vizzaccaro A. , Tosca M. , Passalacqua G. , Pallestrini E. , Canonica G. W. 
Published: June 2005

Background:  Allergic rhinitis is traditionally subdivided into seasonal (SAR) and perennial (PAR), although the new definitions of persistent and intermittent were recently proposed. We assessed the validity of the traditional classification in a large group of subjects suffering from allergic rhinitis alone.Methods:  Young males referred to a Navy Military Hospital for routine fitness visit, and reporting symptoms of rhinitis alone were selected. According to the sensitization they were subdivided into (i) sensitized to pollens only (seasonal, SAR), (ii) to perennial allergens only (perennial, PAR) and (iii) to both (mixed, MAR). Spirometry, methacholine challenge, severity and characteristics of symptoms were assessed in all participants.Results:  Of 19 325 subjects, 2347 had allergic rhinitis. Seventy‐two percent of the subjects had MAR, 17% SAR and 11% PAR. Ocular involvement and irritative symptoms were more frequent in SAR (P < 0.03), whereas obstruction was predominant in PAR (P < 0.01). Nasal symptoms varied according to the period of the year in SAR (P < 0.01) and PAR (P < 0.03). An overt bronchial obstruction was detected in 12% of PAR patients, in 7.8% of MAR, and in 4.2% of SAR. forced expiratory volume/1 s was significantly lower during season in SAR patients only (P < 0.05). The FEF25–75 was impaired in 22.5% MAR patients, 21% PAR, and 14% SAR, with a seasonal change in SAR (P < 0.05) and PAR (P < 0.001). Bronchial hyperreactivity was present in 82.2% of PAR, 73.6% of MAR, and 53.5% of SAR, with a seasonal change in SAR (P < 0.001) and MAR (P < 0.05).Conclusions:  This study provides evidence that up to 80% of allergic rhinitics have a mixed form, and SAR and PAR definitions are poorly adherent to real life. Lung involvement is frequent in patients reporting nose symptoms alone.
Aerobiological and immunochemical studies on Carica papaya L. pollen: an aeroallergen from India
Authors: Chakraborty P. , Ghosh D. , Chowdhury I. , Roy I. , Chatterjee S. , Chanda S. , Gupta‐Bhattacharya S. 
Published: June 2005

Background:  Carica papaya L. is a fruit yielding tree, wildly grown or cultivated in the tropics and subtropics. Its pollen grain has been reported to be airborne and cause immunoglobulin E (IgE)‐mediated hypersensitivity.Objective:  To conduct long‐term aerobiological study on Carica pollen, along with aeroallergenic particles originating from it and to identify vis‐à‐vis characterize an important IgE‐reactive component present in this pollen.Methods:  The seasonal and diurnal periodicities of airborne C. papaya pollen were recorded in a 5‐year survey using a Burkard volumetric sampler. The allergenic potential was studied by skin prick tests, IgE‐enzyme‐linked immunosorbent assay (ELISA) and also by aeroallergen immunoblotting. The total pollen extract was fractionated by Sephacryl S‐200 column, and out of the eluted five fractions, the maximum IgE‐reactive fraction (as found in ELISA inhibition) was resolved into five major subfractions in reverse‐phase high‐performance liquid chromatography (RP‐HPLC). The subfraction with optimum IgE reactivity was studied by activity gel, native and nonreducing sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS‐PAGE). The homogeneity of the isolated protein fraction was checked by crossed immunoelectrophoresis with rabbit antisera and IgE reactivity was confirmed by ELISA inhibition and immunoblotting using individual patient sera.Results:  The Carica pollen occurred in the air round the year with peaks during January and September–October. Among a patient population of 1000, skin‐test results showed 27.8% +1 level and 5.6% +2/+3 level reactions. In aeroallergen immunoblotting of exposed Burkard tape segments, the detected allergen spots showed a significant correlation with airborne pollen count recorded. The pollen extract elicited loss of IgE reactivity when treated with reducing agent‐like β‐mercaptoethanol and heat, but showed six IgE‐reactive components in nonreducing IgE‐immunoblot. The fraction 1 eluted from Sephacryl S‐200 column showed highest IgE reactivity and resolved into five major components in RP‐HPLC. Out of these, the fraction showing optimum IgE reactivity in IgE‐ELISA inhibition and immunoblotting with patient antisera, elicited esterase activity and found to be a homogenous protein of 100 kDa.Conclusion:  Carica papaya tree contributes significantly to the aeropollen and aeroallergen load of the suburban outskirts of Calcutta metropolis, India. The pollen extract contains an important IgE‐reactive protein component of 100 kDa molecular weight with esterase activity.
Molecular cloning and characterization of full‐length cDNAs encoding a novel high‐molecular‐weight Dermatophagoides pteronyssinus mite allergen, Der p 11
Authors: Tsai L. C. , Peng H. J. , Lee C. S. , Chao P. L. , Tang R. B. , Tsai J. J. , Shen H. D. , Hung M. W. , Han S. H. 
Published: June 2005

Background:  Dermatophagoides pteronyssinus (Dp) and D. farinae (Df) mites are the most important source of indoor aeroallergens. Most Dp mite allergens identified to date have relatively low molecular weights (MWs). Identification of high‐MW mite allergens is a crucial step in characterizing the complete spectrum of mite allergens and to provide appropriate tools for diagnostic and therapeutic application.Methods:  The full‐length Der p 11 cDNA clone was isolated using cDNA library immunoscreening, the 5′–3′ rapid amplification of cDNA ends (RACE) system and polymerase chain reactions (PCR). The whole cDNA insert and its PCR‐derived DNA fragments (p1 to p4) were generated and expressed in the Escherichia coli expression system. The allergenicity of the recombinant protein and its peptide fragments was examined by IgE immunodot assays. The IgE‐binding reactivity of rDer p 11 was analyzed in the serum of 50 asthmatic children with positive reactivity to Dp mite extract. Its recombinant peptide fragments were also examined by immunodot assays in 30 mite‐allergic children.Results:  Der p 11 cDNA consists of a 2625‐bp open reading frame encoding a 103‐kDa protein with 875 amino acids. It exhibits significant homology with the paramyosin of other invertebrates. The protein sequence alignment of this newly identified Dp mite allergen (denominated as Der p 11) revealed over 89% identity with Der f 11 and Blo t 1. Among 50 Dp‐sensitive asthmatic children, rDer p 11 showed positive IgE‐binding reactivity to 39 patients (78%). Using immunodot assays, multiple human IgE‐binding activities were demonstrated in all four fragments of Der p 11. Using immunoblot assays, the dominant IgG‐binding epitope for monoclonal antibody (mAb642) was located in fragment p3 only. In immunoblot assays, cross‐inhibition between rDer p 11 and rDer f 11 was up to 73–80% at concentrations of 100 μg/ml.Conclusions:  This study confirms that the newly identified recombinant Der p 11 is a novel and important high‐MW Dp mite allergen for asthmatic children. Our data also indicates that human IgE‐binding major epitopes are scattered over the entire molecule of Der p 11.
Humoral and cellular responses to cow milk proteins in patients with milk‐induced IgE‐mediated and non‐IgE‐mediated disorders
Authors: Shek L. P. C. , Bardina L. , Castro R. , Sampson H. A. , Beyer K. 
Published: June 2005

Background:  Cow milk allergy (CMA) is one of the most common food allergies in childhood. Patients with CMA present with a wide range of immunoglobulin (Ig)E‐ and non‐IgE‐mediated clinical syndromes. Limited information is known about the specific humoral and cellular responses to cow milk proteins in these various forms of CMA.Objective:  The aim of the study was to determine IgE, IgA, IgG1 and IgG4 antibody levels and lymphocyte proliferative responses to the major cow milk allergens in patients with IgE‐ and non‐IgE‐mediated CMA.Methods:  One hundred and forty cow milk allergic patients, 6 months to 22 years of age, were included in the study. One hundred and thirteen patients had IgE‐mediated CMA, 11 had milk protein‐induced enterocolitis syndrome and 16 had allergic eosinophilic gastroenteritis. Twenty‐one patients without food allergy, 8 months to 18 years of age, served as controls. Serum IgE, IgA, IgG1 and IgG4 antibodies to α‐, β‐, and κ‐casein, α‐lactalbumin and β‐lactoglobulin were measured using enzyme‐linked immunosorbent assays. For a subset of these patients, we performed lymphocyte proliferation assays to the various milk allergens.Results:  Patients with IgE‐mediated CMA had higher specific IgE concentrations to casein compared with whey proteins (P < 0.001). In this group of patients, there was a positive correlation between IgE levels and levels of the other isotypes for all four milk proteins (P < 0.001). In general, the caseins were the more allergenic and antigenic proteins in all groups of patients. Patients with enterocolitis syndrome produced less milk protein‐specific IgG4 (P < 0.05) and had a trend for higher IgA antibody levels when compared to the control group. Lymphocyte proliferative responses in all groups with CMA were significantly higher than controls (P < 0.05), although this response was similar in patients with IgE‐ and non‐IgE‐mediated CMA.Conclusion:  There is a distinct pattern of humoral antibody response in the different forms of CMA. Patients with IgE‐mediated CMA have an elevated polyisotypic response to cow milk protein. The relative lack of specific IgG4 production in patients with enterocolitis syndrome may be involved in the pathogenesis of the disease. In general, caseins appear to be the predominant allergen in patients with CMA.
The information needs and preferred roles in treatment decision‐making of parents caring for infants with atopic dermatitis: a qualitative study
Authors: Gore C. , Johnson R. J. , Caress A. L. , Woodcock A. , Custovic A. 
Published: June 2005

Background:  Information needs and preferences in treatment decision‐making of parents caring for infants with atopic dermatitis (AD) are unknown, despite emphasis on quality information‐giving and involvement of health‐care users in treatment decisions.Objective:  To explore information needs and decisional role‐preferences of parents caring for infants with AD.Methods:  Qualitative study. Purposive sample: 31 parents caring for infants with AD. Tape‐recorded focussed conversation‐style interviews. Interview topic‐guide literature‐derived. Control Preferences Scale (5 sort‐card vignettes ‘very active’ to ‘very passive’ role) adapted for use with parents; used to facilitate discussion. Thematic analysis of verbatim transcripts.Results:  Nine core information needs identified: AD‐causation, role of diet, medication‐use, medication‐side‐effects, exacerbating factors, new/alternative medication, nonpharmacological treatments, AD‐prevention, AD and other atopic conditions. Parents desired verbal and written information. Many felt their baby's condition was not taken seriously, leading to delayed diagnosis and treatment. They had to be more active than they wished to obtain information/treatment. Parents preferred sharing decisions with their doctor.Conclusions:  Parents caring for infants with AD have clearly defined, unmet information needs, forcing them into more active roles in the treatment decision‐making process than they desire. The study‐findings may inform the development of written information specifically for these parents and improve partnership during consultations.
Oral food challenges in children in Italy
Authors: Martelli A. , Bouygue G. R. , Isoardi P. , Marelli O. , Sarratud T. , Fiocchi A. 
Published: June 2005

Background:  Epidemiological surveys identify a 900 000‐strong potential paediatric population who may self‐report with food allergy in Italy. As the reference test is the double‐blind, placebo‐controlled food challenge for diagnosis, we carried out a nation‐wide questionnaire survey of hospital‐based paediatric allergy centres between 1 January and 30 June 2003.Methods:  All registered centres received a 27‐point questionnaire polling logistics and protocols of oral food challenge. Items polled included medical facilities and personnel, selection of patients, foods tested, blinding, placebo use, evaluation criteria, number of tests and positive outcome rates in 2002.Results:  Two hundred and two of 456 responders (44.3%) carried out food challenges (a ratio of one centre to 37 400 children in the general population). 11 728 challenges were performed in 2002 (an average of 58 procedures per centre; range: 1–500) and 3350 procedures were claimed positive (28.6%). Cow's milk was tested by all centres, egg by 77.2%, beef by 19.8% and cereals, peanuts, fish or fruit collectively by 37.6%. Twenty‐nine of 202 centres performed blinded tests, 18 of which under double‐blinded conditions. In cow's milk challenges, placebos included lyophilized cow's milk capsules, soy formula, extensive whey hydrolysate or extensive casein hydrolysate.Conclusion:  Our survey reveals great differences in challenge test criteria, facilities and procedures and that many centres perform relatively few procedures. This argues in favour of greater rationalization in the allocation of tertiary‐level resources before procedure standardization can be contemplated in the diagnostic workup of food allergy.
Human hair is a potential source of cat allergen contamination of ambient air
Authors: Karlsson A.‐S. , Renström A. 
Published: June 2005

Background:  We have previously shown that airborne cat allergen levels are significantly lower in school classes using special school clothing or in classes with no pet owners. However, cat allergen is present and the levels are in fact two‐ to threefold higher on cat owners’ than noncat owners’ school clothing which is used, washed and stored at school only. This suggests that allergen is transferred to schools by routes other than clothing.Aim:  To analyse levels of cat allergen (Fel d 1) in hair from cat owners and noncat owners among children and adults.Methods:  Samples of unwashed hair (≥1 day prior to sampling) from adults and children with (n = 22) or without (n = 22) cats at home were collected at a hairdresser. In addition, samples of newly washed hair (adults only, n = 11) were collected. The hair sample was extracted and analysed for Fel d 1 content with ELISA.Results:  The geometric mean levels were more than two orders of magnitude higher in unwashed hair from cat owners, compared with noncat owners (P < 0.0001) and more than 10‐fold higher in newly washed hair from adults. The allergen contamination of unwashed hair among noncat owners appeared higher in children than in adults (P = 0.045).Conclusions:  Hair may be an important source for transfer and deposition of cat allergen in schools and may explain why cat allergen is found in environments with strict allergen avoidance measures. Although it may be unrealistic to apply allergen avoidance strategies against this allergen source, it is important to be aware of it.
Intranasal and inhaled fluticasone propionate for pollen‐induced rhinitis and asthma
Authors: Dahl R. , Nielsen L. P. , Kips J. , Foresi A. , van Cauwenberge P. , Tudoric N. , Howarth P. , Richards D. H. , Williams M. , Pauwels R. ,   the SPIRA Study Group
Published: June 2005

Background:  Studies suggest that nasal treatment might influence lower airway symptoms and function in patients with comorbid rhinitis and asthma. We investigated the effect of intranasal, inhaled corticosteroid or the combination of both in patients with both pollen‐induced rhinitis and asthma.Methods:  A total of 262 patients were randomized to 6 weeks’ treatment with intranasal fluticasone propionate (INFP) 200 μg o.d., inhaled fluticasone propionate (IHFP) 250 μg b.i.d., their combination, or intranasal or inhaled placebo, in a multicentre, double‐blind, parallel‐group study. Treatment was started 2 weeks prior to the pollen season and patients recorded their nasal and bronchial symptoms twice daily. Before and after 4 and 6 weeks’ treatment, the patients were assessed for lung function, methacholine responsiveness, and induced sputum cell counts.Results:  Intranasal fluticasone propionate significantly increased the percentages of patients reporting no nasal blockage, sneezing, or rhinorrhoea during the pollen season, compared with IHFP or intranasal or inhaled placebo. In contrast, only IHFP significantly improved morning peak‐flow, forced expiratory volume in 1 second (FEV1) and methacholine PD20, and the seasonal increase in the sputum eosinophils and methacholine responsiveness.Conclusions:  In patients with pollen‐induced rhinitis and asthma, the combination of intranasal and IHFP is needed to control the seasonal increase in nasal and asthmatic symptoms.