bluuebunny/crossref_metadata_2025_split huggingface dataset:
Isolation and Characterization of the Neutral Leucine Aminopeptidase (LapN) of Tomato
Authors: Tu Chao-Jung , Park Sang-Youl , Walling Linda L. 
Published: May 2003

Abstract
Tomatoes (Lycopersicon esculentum) express two forms of leucine aminopeptidase (LAP-A and LAP-N) and two LAP-like proteins. The relatedness of LAP-N and LAP-A was determined using affinity-purified antibodies to four LAP-A protein domains. Antibodies to epitopes in the most N-terminal region were able to discriminate between LAP-A and LAP-N, whereas antibodies recognizing central and COOH-terminal regions recognized both LAP polypeptides. Two-dimensional immunoblots showed that LAP-N and the LAP-like proteins were detected in all vegetative (leaves, stems, roots, and cotyledons) and reproductive (pistils, sepals, petals, stamens, and floral buds) organs examined, whereas LAP-A exhibited a distinct expression program.LapN was a single-copy gene encoding a rare-class transcript. A full-length LapN cDNA clone was isolated, and the deduced sequence had 77% peptide sequence identity with the wound-induced LAP-A. Comparison of LAP-N with other plant LAPs identified 28 signature residues that classified LAP proteins as LAP-N or LAP-A like. Overexpression of a His6-LAP-N fusion protein in Escherichia coli demonstrated distinct differences in His6-LAP-N and His6-LAP-A activities. Similar to LapA, the LapN RNA encoded a precursor protein with a molecular mass of 60 kD. The 5-kD presequence had features similar to plastid transit peptides, and processing of the LAP-N presequence could generate the mature 55-kD LAP-N. Unlike LapA, the LapN transcript contained a second in-frame ATG, and utilization of this potential initiation codon would yield a 55-kD LAP-N protein. The localization of LAP-N could be controlled by the balance of translational initiation site utilization and LAP-N preprotein processing.
Enhancement of Plant-Microbe Interactions Using a Rhizosphere Metabolomics-Driven Approach and Its Application in the Removal of Polychlorinated Biphenyls,
Authors: Narasimhan Kothandaraman , Basheer Chanbasha , Bajic Vladimir B. , Swarup Sanjay 
Published: May 2003

Abstract
Persistent organic pollutants, such as polychlorinated biphenyls (PCBs), are a global problem. We demonstrate enhanced depletion of PCBs using root-associated microbes, which can use plant secondary metabolites, such as phenylpropanoids. Using a “rhizosphere metabolomics” approach, we show that phenylpropanoids constitute 84% of the secondary metabolites exuded from Arabidopsis roots. Phenylpropanoid-utilizing microbes are more competitive and are able to grow at least 100-fold better than their auxotrophic mutants on roots of plants that are able to synthesize or overproduce phenylpropanoids, such as flavonoids. Better colonization of the phenylpropanoid-utilizing strain in a gnotobiotic system on the roots of flavonoid-producing plants leads to almost 90% removal of PCBs in a 28-d period. Our work complements previous approaches to engineer soil microbial populations based on opines produced by transgenic plants and used by microbes carrying opine metabolism genes. The current approach based on plant natural products can be applied to contaminated soils with pre-existing vegetation. This strategy is also likely to be applicable to improving the competitive abilities of biocontrol and biofertilization strains.
Nod Factor and Elicitors Activate Different Phospholipid Signaling Pathways in Suspension-Cultured Alfalfa Cells
Authors: den Hartog Martine , Verhoef Nathalie , Munnik Teun 
Published: May 2003

Abstract
Lipo-chitooligosaccharides (Nod factors) are produced by symbiotic Rhizobium sp. bacteria to elicit Nod responses on their legume hosts. One of the earliest responses is the formation of phosphatidic acid (PA), a novel second messenger in plant cells. Remarkably, pathogens have also been reported to trigger the formation of PA in nonlegume plants. To investigate how host plants can distinguish between symbionts and pathogens, the effects of Nod factor and elicitors (chitotetraose and xylanase) on the formation of PA were investigated in suspension-cultured alfalfa (Medicago sativa) cells. Theoretically, PA can be synthesized via two signaling pathways, i.e. via phospholipase D (PLD) and via phospholipase C in combination with diacylglycerol (DAG) kinase. Therefore, a strategy involving differential radiolabeling with [32P]orthophosphate was used to determine the contribution of each pathway to PA formation. In support, PLD activity was specifically measured by using the ability of the enzyme to transfer the phosphatidyl group of its substrate to a primary alcohol. In practice, Nod factor, chitotetraose, and xylanase induced the formation of PA and its phosphorylated product DAG pyrophosphate within 2 min of treatment. However, whereas phospholipase C and DAG kinase were activated during treatment with all three different compounds, PLD was only activated by Nod factor. No evidence was obtained for the activation of phospholipase A2.
Growth Ring Formation in the Starch Granules of Potato Tubers
Authors: Pilling Emma , Smith Alison M. 
Published: May 2003

Abstract
Starch granules from higher plants contain alternating zones of semicrystalline and amorphous material known as growth rings. The regulation of growth ring formation is not understood. We provide several independent lines of evidence that growth ring formation in the starch granules of potato (Solanum tuberosum) tubers is not under diurnal control. Ring formation is not abolished by growth in constant conditions, and ring periodicity and appearance are relatively unaffected by a change from a 24-h to a 40-h photoperiod, and by alterations in substrate supply to the tuber that are known to affect the diurnal pattern of tuber starch synthesis. Some, but not all, of the features of ring formation are consistent with the involvement of a circadian rhythm. Such a rhythm might operate by changing the relative activities of starch-synthesizing enzymes: Growth ring formation is disrupted in tubers with reduced activity of a major isoform of starch synthase. We suggest that physical as well as biological mechanisms may contribute to the control of ring formation, and that a complex interplay of several factors may by involved.
Solubilization of an Arabinan Arabinosyltransferase Activity from Mung Bean Hypocotyls
Authors: Nunan Kylie Joy , Scheller Henrik Vibe 
Published: May 2003

Abstract
The biosynthesis of polysaccharides destined for the plant cell wall and the subsequent assembly of the cell wall are poorly understood processes that are currently the focus of much research. Arabinan, a component of the pectic polysaccharide rhamnogalacturonan I, is composed of arabinosyl residues connected via various glycosidic linkages, and therefore, the biosynthesis of arabinan is likely to involve more than one arabinosyltransferase. We have studied the transfer of [14C]arabinose (Ara) from UDP-l-arabinopyranose onto polysaccharides using microsomal membranes isolated from mung bean (Vigna radiata) hypocotyls. [14C]arabinosyl and [14C]xylosyl residues were incorporated into endogenous products due to the presence of UDP-Xyl-4-epimerase activity. Enzymatic digestion of endogenous products with endo-arabinanase released very little radiolabeled sugars, whereas digestion with arabinofuranosidase released some [14C]Ara. Microsomal membranes solubilized with the detergent octyl glucoside were able to add a single [14C]Ara residue onto (1→5)-linked α-l-arabino-oligosaccharide acceptors. The reaction had a pH optimum of 6.5 and a requirement for manganese ions. However, enzymatic digestion of the radiolabeled oligosaccharides with endo-arabinanase and arabinofuranosidases could not fully release the radiolabeled Ara residue, indicating that the [14C]Ara residue was not a (1→2)-, (1→3)-, or (1→5)-linked α-l-arabinofuranosyl residue. Rather, mild acid treatment of the product suggested that the radiolabeled Ara residue was in a pyranose conformation, and this result was confirmed by thin-layer chromatography of radiolabeled partially methylated sugars. Using microsomal membranes separated on a discontinuous sucrose gradient, the arabinosyltransferase activity appears to be mainly localized to Golgi membranes.
Molecular Cloning and Functional Characterization of Three Distinct N-Methyltransferases Involved in the Caffeine Biosynthetic Pathway in Coffee Plants
Authors: Uefuji Hirotaka , Ogita Shinjiro , Yamaguchi Yube , Koizumi Nozomu , Sano Hiroshi 
Published: May 2003

Abstract
Caffeine is synthesized from xanthosine throughN-methylation and ribose removal steps. In the present study, three types of cDNAs encodingN-methyltransferases were isolated from immature fruits of coffee (Coffea arabica) plants, and designated asCaXMT1, CaMXMT2, andCaDXMT1, respectively. The bacterially expressed encoded proteins were characterized for their catalytic properties. CaXMT1 catalyzed formation of 7-methylxanthosine from xanthosine with aK m value of 78 μm, CaMXMT2 catalyzed formation of 3,7-dimethylxanthine (theobromine) from 7-methylxanthine with a K m of 251 μm, and CaDXMT1 catalyzed formation of 1,3,7-trimethylxanthine (caffeine) from 3,7-dimethylxanthine with aK m of 1,222 μm. The crude extract of Escherichia coli was found to catalyze removal of the ribose moiety from 7-methylxanthosine, leading to the production of 7-methylxanthine. As a consequence, when all three recombinant proteins and E. coli extract were combined, xanthosine was successfully converted into caffeine in vitro. Transcripts for CaDXMT1 were predominantly found to accumulate in immature fruits, whereas those for CaXMT1and CaMXMT2 were more broadly detected in sites encompassing the leaves, floral buds, and immature fruits. These results suggest that the presently identified threeN-methyltransferases participate in caffeine biosynthesis in coffee plants and substantiate the proposed caffeine biosynthetic pathway: xanthosine → 7-methylxanthosine → 7-methylxanthine → theobromine → caffeine.
SEMICONDUCTOR THIN FILMS AND THIN FILM DEVICES FOR ELECTROTEXTILES
Authors: SHUR MICHAEL S. , RUMYANTSEV SERGEY L. , GASKA REMIS 
Published: May 2003

We discuss the evolution from wearable electronics and conductive textiles to electrotextiles with embedded semiconducting films and semiconductor devices and review different semiconductor technologies competing for applications in electrotextiles. We also report on fabrication, characterization, and properties of nanocrystalline semiconductor and metal films and thin-film device structures chemically deposited on fibers, cloth, and large area flexible substrates at low temperatures (close to room temperature). Our approach is based on a new process of depositing polycrystalline CdSe (1.75 eV), CdS (2.4 eV), PbS (0.4 eV), PbSe (0.24 eV) and CuxS (semiconductor/metal) films on flexible substrates from the water solutions of complex-salt compounds. We have covered areas up to 8 × 10 inches but the process can be scaled up. The film properties are strongly affected by processing. We fabricated a lateral solar cell with alternating Cu2-xS and nickel contact stripes deposited on top of a view foil. These sets of contacts represented "ohmic" and "non-ohmic" contacts, respectively. Then CdS films of approximately 0.5 μm thick were deposited on top. We also fabricated a "sandwich" type photovoltaic cell, where the CdS film was sandwiched between an In2O3 layer deposited on a view foil and a Cu2-xS layer deposited on top. Both structures exhibited transient response under light, with the characteristic response time decreasing with the illumination wavelength. This is consistent with having deeper localized states in the energy gap determining the transients for shorter wavelength radiation. (Slow transients related to trapping effects are typical for polycrystalline CdS materials.) We also report on the photovoltaic effect in CdS/CuS films deposited on trylene threads and on a field effect in these films deposited on a flexible copper wire. CdS films deposited on viewfoils exhibit unique behavior under stress and UV radiation exposure with reproducible resistance changes of several orders of magnitude with bending up to 10 mm curvature. Our results clearly demonstrate the feasibility of using this technology for photovoltaic and microelectronics applications for electrotextiles and wearable electronics applications.
Reasoning, Models, and Images: Behavioral Measures and Cortical Activity
Authors: Knauff Markus , Fangmeier Thomas , Ruff Christian C. , Johnson-Laird P. N. 
Published: May 2003

Abstract
The goal of this study was to investigate the neurocognitive processes of mental imagery in deductive reasoning. Behavioral studies yielded four sorts of verbal relations: (1) visuospatial relations that are easy to envisage both visually and spatially; (2) visual relations that are easy to envisage visually but hard to envisage spatially; (3) spatial relations that are hard to envisage visually but easy to envisage spatially; and (4) control relations that are hard to envisage both visually and spatially. In three experiments, visual relations slowed the process of reasoning in comparison with control relations, whereas visuospatial and spatial relations yielded inferences comparable to those of control relations. An experiment using functional magnetic resonance imaging showed that in the absence of any correlated visual input (problems were presented acoustically via headphones), all types of reasoning problems evoked activity in the left middle temporal gyrus, in the right superior parietal cortex, and bilaterally in the precuneus. In the prefrontal cortex, increased activity was found in the middle and inferior frontal gyri. However, only the problems based on visual relations also activated areas of the visual association cortex corresponding to V2. The results indicate that cortical activity during reasoning depends on the nature of verbal relations. All relations elicit mental models that underlie reasoning, but visual relations in addition elicit visual images. This account resolves inconsistencies in the previous literature.
Human Cerebellum: Surface-Assisted Cortical Parcellation and Volumetry with Magnetic Resonance Imaging
Authors: Makris Nikos , Hodge Steven M. , Haselgrove Christian , Kennedy David N. , Dale Anders , Fischl Bruce , Rosen Bruce R. , Harris Gordon , Caviness Verne S. , Schmahmann Jeremy D. 
Published: May 2003

Abstract
We describe a system of surface-assisted parcellation (SAP) of the human cerebellar cortex derived from neural systems functional and behavioral anatomy. This system is based on MRI and preserves the unique morphologic and topographic features of the individual cerebellum. All major fissures of the cerebellum were identified and traced in the flattened representation of the cerebellar cortex using the program “Free Surfer.” Parcellation of the cerebellar cortex followed using the fissure information in conjunction with landmarks using the program “Cardviews” to create 64 gyral-based cerebellar parcellation units. Computer-assisted algorithms enable the execution of the cerebellar parcellation procedure as well as volumetric measurements and topographic localization. The SAP technique makes it possible to represent multimodal structural and functional imaging data on the flattened surface of the cerebellar cortex as illustrated in one functional MRI experiment.
Drug-induced thrombocytopenia: localization of the binding site of GPIX-specific quinine-dependent antibodies
Authors: Asvadi Parisa , Ahmadi Zohra , Chong Beng H. 
Published: May 2003

AbstractImmune thrombocytopenia is a common complication of therapy with a large number of drugs. The most widely studied drug-induced immune thrombocytopenia (DIT) is caused by quinine. In most cases of DIT, antibodies bind to the platelet membrane glycoprotein (GP) Ib-IX complex in a drug-dependent fashion and bring about increased platelet clearance by the reticuloendothelial system resulting in thrombocytopenia. Here, we report the characterization of the quinine-dependent antibody activity of sera from 13 patients with quinine-induced thrombocytopenia. In our series of patients, GPIX was the most prevalent target of quinine-dependent antibodies. To identify the structural determinants of GPIX recognized by quinine-dependent antibodies, 4 chimeric mouse/human GPIX constructs and stable Chinese hamster ovary (CHO) cell lines that expressed the chimeras in association with GPIbα and GPIbβ were produced. The analysis of 6 patient sera with the chimeric cell lines provided evidence for localization of the anti-GPIX quinine-dependent antibody binding site to the C-ext region (amino acid [aa] 64-135) of human GPIX. Further characterization of the C-ext region of the GPIX indicated that replacement of the Arg110 and Gln115 of the human GPIX with the corresponding residues from mouse (Gln and Glu, respectively) resulted in a significant reduction in the binding of GPIX antibodies in our series of patients, with Arg110Gln, giving a more pronounced effect than Gln115Glu. Hence, these 2 residues, particularly Arg110, play an important role in the structure of the antigenic site on GPIX recognized by anti-GPIX antibodies.
Mithramycin induces fetal hemoglobin production in normal and thalassemic human erythroid precursor cells
Authors: Fibach Eitan , Bianchi Nicoletta , Borgatti Monica , Prus Eugenia , Gambari Roberto 
Published: May 2003

AbstractWe report in this paper that the DNA-binding drug mithramycin is a potent inducer of γ-globin mRNA accumulation and fetal hemoglobin (HbF) production in erythroid cells from healthy human subjects and β-thalassemia patients. Erythroid precursors derived from peripheral blood were grown in 2-phase liquid culture. In this procedure, early erythroid progenitors proliferate and differentiate during phase 1 (in the absence of erythropoietin) into late progenitors. In phase 2, in the presence of erythropoietin, the latter cells continue their proliferation and mature into Hb-containing orthochromatic normoblasts. Compounds were added on days 4 to 5 of phase 2 (when cells started to synthesize Hb), and cells were harvested on day 12. Accumulation of mRNAs for γ-globin, β-globin, α-globin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and β-actin were measured by real-time quantitative reverse transcription–polymerase chain reaction (RT-PCR); induction of HbF was analyzed by high-performance liquid chromatography (HPLC) and, at cellular level, by flow cytometry. We demonstrated that mithramycin was able to up-regulate preferentially γ-globin mRNA production and to increase HbF accumulation, the percentage of HbF-containing cells, and their HbF content. Mithramycin was more effective than hydroxyurea, being, in addition, not cytotoxic. This was shown by the lack of cytotoxicity on erythroid and myeloid in vitro primary cell cultures treated with mithramycin at concentrations effective for HbF induction. These results are of potential clinical significance because an increase of HbF alleviates the symptoms underlying β-thalassemia and sickle cell anemia. The results of this report suggest that mithramycin and its analogs warrant further evaluation as potential therapeutic drugs.
Sustained phenotypic correction of canine hemophilia A using an adeno-associated viral vector
Authors: Scallan Ciaran D. , Lillicrap David , Jiang Haiyan , Qian Xiaobing , Patarroyo-White Susannah L. , Parker Amy E. , Liu Tongyao , Vargas Joseph , Nagy Dea , Powell Sharon K. , Wright J. Fraser , Turner Patricia V. , Tinlin Shawn J. , Webster Sandra E. , McClelland Alan , Couto Linda B. 
Published: May 2003

Abstract
Gene therapy for hemophilia A requires efficient delivery of the factor VIII gene and sustained protein expression at circulating levels of at least 1% to 2% of normal. Adeno-associated viral type 2 (AAV2) vectors have a number of advantages over other viral vectors, including an excellent safety profile and persistent gene expression. However, a major disadvantage is their small packaging capacity, which has hampered their use in treating diseases such as hemophilia A, cystic fibrosis, and muscular dystrophy, which are caused by mutations in large genes. Here we demonstrate that this can be overcome by using small regulatory elements to drive expression of a B-domain–deleted form of FVIII. The use of this vector for hepatic gene transfer in a canine model of hemophilia A resulted in the sustained (&gt; 14 months) expression of biologically active FVIII. FVIII activity levels of 2% to 4% were achieved. These levels correlated with a partial correction in the whole-blood clotting time and cuticle bleeding time. In addition, immunoprecipitation analysis demonstrated the expression of canine FVIII of the predicted size in the plasma of injected animals. These data support the use of AAV2 vectors in human clinical trials to treat hemophilia A patients.
Conventional or physicochemical approach in intensive care unit patients with metabolic acidosis
Authors: Moviat MAM , van Haren FMP , van der Hoeven JG 
Published: May 2003

Abstract
Introduction
Metabolic acidosis is the most frequent acid–base disorder in the intensive care unit. The optimal analysis of the underlying mechanisms is unknown.

Aim
To compare the conventional approach with the physicochemical approach in quantifying complicated metabolic acidosis in patients in the intensive care unit

Patients and methods
We included 50 consecutive patients with a metabolic acidosis (standard base excess ≤ -5). We measured sodium, potassium, calcium, magnesium, chloride, lactate, creatinine, urea, phosphate, albumin, pH, and arterial carbon dioxide and oxygen tensions in every patient. We then calculated HCO3-, the base excess, the anion gap, the albumin-corrected anion gap, the apparent strong ion difference, the effective strong ion difference and the strong ion gap.

Results
Most patients had multiple underlying mechanisms explaining the metabolic acidosis. Unmeasured strong anions were present in 98%, hyperchloremia was present in 80% and elevated lactate levels were present in 62% of patients. Calculation of the anion gap was not useful for the detection of hyperlactatemia. There was an excellent relation between the strong ion gap and the albumin-corrected and lactate-corrected anion gap (r2 = 0.934), with a bias of 1.86 and a precision of 0.96.

Conclusion
Multiple underlying mechanisms are present in most intensive care unit patients with a metabolic acidosis. These mechanisms are reliably determined by measuring the lactate-corrected and albumin-corrected anion gap. Calculation of the more time-consuming strong ion gap according to Stewart is therefore unnecessary.