bluuebunny/crossref_metadata_2025_split huggingface dataset:
A treatment trade‐off based decision aid for patients with locally advanced non‐small cell lung cancer
Authors: Brundage Michael D. , Feldman‐Stewart Deb , Dixon Peter , Gregg Richard , Youssef Youssef , Davies Diane , Mackillop William J. 
Published: March 2003

Purpose To describe the structure and use of a decision aid for patients with locally advanced non‐small cell lung cancer (LA‐NSCLC) who are eligible for combined‐modality treatment (CMT) or for radiotherapy alone (RT).Methods The aid included a structured description of the treatment options and trade‐off exercises designed to help clarify the patient’s values for the relevant outcomes by determining the patient’s survival advantage threshold (the increase in survival conferred by CMT over RT that the patient deemed necessary for choosing CMT). Additional outcome measures included each patient’s strength of treatment preference, decisional conflict, objective understanding of survival information, decisional role preference, and evaluation of the aid itself.Results Twenty‐five patients met the eligibility criteria for study. Of these, seven declined the decision aid because they had a clear treatment preference (four chose CMT and three chose RT). The remaining 18 participants completed the decision aid; 16 chose CMT and two chose RT. All 18 patients wished to participate in the decision to some extent. All patients reported that using the decision support was useful to them and recommended its use for others. No patient or physician reported that the aid interfered with the physician‐patient relationship. Patients’ 3‐year survival advantage thresholds, and their median survival advantage thresholds, were each strongly correlated with their strengths of treatment preference (ρ=0.80,P < 0.001 andρ=0.77,P < 0.001, respectively). For all but one patient, either their 3‐year or median survival threshold was consistent with their final treatment choice. Eight patients reported a stronger treatment preference after using the decision aid.Conclusions We conclude that a treatment trade‐off based decision aid for patients with locally advanced non‐small cell lung cancer is feasible, that it demonstrates internal consistency and convergent validity, and that it is favourably evaluated by patients and their physicians. The aid seems to help patients understand the benefits and risks of treatment and to choose the treatment that is most consistent with their values.
Patients' attitudes to medicines and expectations for prescriptions
Authors: Britten Nicky , Ukoumunne Obioha C. , Boulton Mary G. 
Published: March 2003

AbstractBackground  Recent research has shown that patients' expectations for prescriptions influence doctors' prescribing decisions, but little is known of the antecedents of these expectations.Objectives  To test earlier qualitative research about patients' views of medicines; to describe the demographic characteristics of those holding orthodox and unorthodox views of medicines; to investigate the relationship between patients' ideal and predicted expectations for prescriptions; and to determine the relative effects of attitudinal, demographic, organizational and illness variables on these expectations.Design  Questionnaire survey of patients consulting general practitioners.Setting and participants  A total of 544 patients and 15 doctors in four general practices.Main variables studied  Patients' attitudes to medicines; patients' demographic characteristics; organizational variables; aspects of patients' presenting problems.Outcome measures  Patients' ideal and predicted expectations for prescriptions.Results  Orthodox and unorthodox attitudes to medicines can be measured quantitatively, and ethnicity was the only demographic variable associated with both. Ideal and predicted expectations for prescriptions were closely related to each other but differed in their antecedents. Both types of expectations were associated with attitudinal, demographic, organizational and illness variables. Ideal expectations were influenced by orthodox and unorthodox attitudes to medicines, while predicted expectations were only influenced by orthodox attitudes.Conclusions  Future studies of patients' expectations for health services should distinguish between ideal and predicted expectations, and should consider the range of possible influences on these expectations. In particular, the effect of the organization and context of health services should be investigated.
Excessive vitamin A toxicity in mice genetically deficient in either alcohol dehydrogenase Adh1 or Adh3
Authors: Molotkov Andrei , Fan Xiaohong , Duester Gregg 
Published: March 2003

Alcohol dehydrogenase (ADH) deficiency results in decreased retinol utilization, but it is unclear what physiological roles the several known ADHs play in retinoid signaling. Here, Adh1, Adh3, and Adh4 null mutant mice have been examined following acute and chronic vitamin A excess. Following an acute dose of retinol (50 mg·kg−1), metabolism of retinol to retinoic acid in liver was reduced 10‐fold in Adh1 mutants and 3.8‐fold in Adh3 mutants, but was not significantly reduced in Adh4 mutants. Acute retinol toxicity, assessed by determination of the LD50 value, was greatly increased in Adh1 mutants and moderately increased in Adh3 mutants, but only a minor effect was observed in Adh4 mutants. When mice were propagated for one generation on a retinol‐supplemented diet containing 10‐fold higher vitamin A than normal, Adh3 and Adh4 mutants had essentially the same postnatal survival to adulthood as wild‐type (92–95%), but only 36% of Adh1 mutants survived to adulthood with the remainder dying by postnatal day 3. Adh1 mutants surviving to adulthood on the retinol‐	supplemented diet had elevated serum retinol signifying a clearance defect and elevated aspartate aminotransferase indicative of increased liver damage. These findings indicate that ADH1 functions as the primary enzyme responsible for efficient oxidative clearance of excess retinol, thus providing protection and increased survival during vitamin A toxicity. ADH3 plays a secondary role. Our results also show that retinoic acid is not the toxic moiety during vitamin A excess, as Adh1 mutants have less retinoic acid production while experiencing increased toxicity.
Functional integration of mitochondrial and hydrogenosomal ADP/ATP carriers in the Escherichia coli membrane reveals different biochemical characteristics for plants, mammals and anaerobic chytrids
Authors: Haferkamp Ilka , Hackstein Johannes H. P. , Voncken Frank G. J. , Schmit Guillaume , Tjaden Joachim 
Published: March 2003

The expression of mitochondrial and hydrogenosomal ADP/ATP carriers (AACs) from plants, rat and the anaerobic chytridiomycete fungus Neocallimastix spec. L2 in Escherichia coli allows a functional integration of the recombinant proteins into the bacterial cytoplasmic membrane. For AAC1 and AAC2 from rat, apparent Km values of about 40 µm for ADP, and 105 µm or 140 µm, respectively, for ATP have been determined, similar to the data reported for isolated rat mitochondria. The apparent Km for ATP decreased up to 10‐fold in the presence of the protonophore m‐chlorocarbonylcyanide phenylhydrazone (CCCP). The hydrogenosomal AAC isolated from the chytrid fungus Neocallimastix spec. L2 exhibited the same characteristics, but the affinities for ADP (165 µm) and ATP (2.33 mm) were significantly lower. Notably, AAC1‐3 from Arabidopsis thaliana and AAC1 from Solanum tuberosum (potato) showed significantly higher external affinities for both nucleotides (10–22 µm); they were only slightly influenced by CCCP.Studies on intact plant mitochondria confirmed these observations. Back exchange experiments with preloaded E. coli cells expressing AACs indicate a preferential export of ATP for all AACs tested. This is the first report of a functional integration of proteins belonging to the mitochondrial carrier family (MCF) into a bacterial cytoplasmic membrane. The technique described here provides a relatively simple and highly reproducible method for functional studies of individual mitochondrial‐type carrier proteins from organisms that do not allow the application of sophisticated genetic techniques.
Biglycan isoforms with differences in polysaccharide substitution and core protein in human lung fibroblasts
Authors: Tufvesson Ellen , Malmström Johan , Marko‐Varga György , Westergren‐Thorsson Gunilla 
Published: March 2003

Biglycan is widely distributed in the extracellular matrix and is a member of the small proteoglycan family characterized by a core protein with leucine‐rich repeat motifs. We show in this paper for the first time that biglycan from human lung fibroblasts can be expressed as different isoforms. These isoforms can be separated from the predominant form of biglycan by hydrophobic interaction chromatography, where the more hydrophobic isoforms are retarded. The newly found isoforms of biglycan have a smaller core protein substituted with smaller glycosaminoglycan chains, migrating on SDS/PAGE at between 110 and 200 kDa. These molecules were identified as biglycan using MALDI‐TOF MS. Identification of C‐terminal peptides together with glycosylation of the N‐terminal glycosaminoglycan sites excludes the possibility of terminal proteolytic cleavage. The biglycan isoforms are N‐glycosylated, which demonstrates that a lack in N‐glycosylation is not the reason for a smaller core. Two components revealed by RT‐PCR indicate alternative splicing, which could be located in regions of the protein that have not been identified, with the exclusion of sites of glycosylations.Analyses of glycosaminoglycan chain length of the isoforms show that besides the normally occurring glycosaminoglycan chains, there is a mixture of shorter glycosaminoglycan chains. Structural analysis shows that these glycosaminoglycan chains contain a lower proportion of iduronic acid (61%) relative to glucuronic acid when compared to the glycosaminoglycan chain of the predominant form of biglycan (71%). We can anticipate that variation in structure of biglycan can cause changes in the connective tissue formation depending on its ability to bind matrix molecules, as well as cytokines.
Directed evolution of a glutaryl acylase into an adipyl acylase
Authors: Sio Charles F. , Riemens Anja M. , van der Laan Jan‐Metske , Verhaert Raymond M.D. , Quax Wim J. 
Published: March 2003

Semi‐synthetic cephalosporin antibiotics belong to the top 10 of most sold drugs, and are produced from 7‐aminodesacetoxycephalosporanic acid (7‐ADCA). Recently new routes have been developed which allow for the production of adipyl‐7‐ADCA by a novel fermentation process. To complete the biosynthesis of 7‐ADCA a highly active adipyl acylase is needed for deacylation of the adipyl derivative. Such an adipyl acylase can be generated from known glutaryl acylases.The glutaryl acylase of Pseudomonas SY‐77 was mutated in a first round by exploration mutagenesis. For selection the mutants were grown on an adipyl substrate. The residues that are important to the adipyl acylase activity were identified, and in a second round saturation mutagenesis of this selected stretch of residues yielded variants with a threefold increased catalytic efficiency. The effect of the mutations could be rationalized on hindsight by the 3D structure of the acylase.In conclusion, the substrate specificity of a dicarboxylic acid acylase was shifted towards adipyl‐7‐ADCA by a two‐step directed evolution strategy. Although derivatives of the substrate were used for selection, mutants retained activity on the β‐lactam substrate. The strategy herein described may be generally applicable to all β‐lactam acylases.
Structure determination of the capsular polysaccharide from Vibrio vulnificus strain 6353
Authors: Reddy G. Prabhakar , Hayat Unaiza , Xu Qiuwei , Reddy K. Vasavi , Wang Yuhui , Chiu Kou‐Wei , Morris Jr J. Glenn , Bush C. Allen 
Published: March 2003

Vibrio vulnificus is a pathogenic gram‐negative bacterium, endemic to brackish waters, which is often isolated from sediments, from the water column or from shellfish. It is associated with wound infections and septicemia in humans and the virulence of V. vulnificus has been strongly associated with encapsulation. The capsular polysaccharide purified from a virulent strain of V. vulnificus 6353 did not show cross reactivity with antibodies to the capsular polysaccharide of a related pathogenic strain of V. vulnificus (MO6‐24) the structure of which was recently reported. NMR spectroscopic analysis of the purified polysaccharide from strain 6353 showed that the polymer is composed of four sugar residues per repeating subunit including 2,6‐dideoxy‐2‐N‐acetylamino‐α‐
D‐glucose (QuiNAc), 2‐deoxy‐2‐N‐acetylamino‐α‐
D‐galactose (α‐
D‐GalNAc), 2‐deoxy‐2‐N‐acetylamino‐α‐
D‐galcturonic acid (α‐
D‐GalNAcA) and 2‐N‐acetylamino‐α‐
D‐glucuronamide (α‐
D‐GlcNAcANH2). The 1H‐ and 13C‐NMR spectra were completely assigned by homonuclear and heteronuclear NMR spectroscopy. Sugar types and anomeric configurations were determined from proton homonuclear coupling constants and glycosidic linkages were determined from 1H−13C heteronuclear multiple bond correlation spectra. Sugar identities were confirmed by high performance anion‐exchange chromatography and absolute configurations were determined by gas chromatography in combination with molecular modeling and NMR spectroscopy. The structure of the polysaccharide repeating unit is :[←4)‐α‐.cf15.D.cf15..cf1.‐Gal.cf2.p.cf2..cf1.NAc‐(1←3)‐α‐.cf15.D.cf15..cf1.‐Gal.cf2.p.cf2..cf1.NAcA‐(1←3)‐α‐.cf15.D.cf15..cf1.‐Qui.cf2.p.cf2..cf1.NAc‐(1←].cf8.n.cf8..cf1.α‐
D‐GlcpNAcANH2 (1←4)‐↗While there are some common features shared among the structures of the capsular polysaccharides of pathogenic strains of V. vulnificus, there are distinct differences in the detailed structures.
Molecular and pharmacological characterization of recombinant rat/mice N‐methyl‐	D‐aspartate receptor subtypes in the yeast Saccharomyces cerevisiae
Authors: Becker Jürgen , Li Ziyu , Noe Christian R. 
Published: March 2003

The genes encoding the ionotropic N‐methyl‐
D‐aspartate (NMDA) receptor subunits NR1a, NR2B and NR2D were cloned in the multi‐copy yeast‐Escherichia coli shuttle vectors pMB01 and pMB02. The protease‐deficient yeast Saccharomyces cerevisiae cl3‐ABYS‐86 (leu−, ura−, his−) was transformed with the recombinant plasmids pMBNR1a (leu+), pMBNR1a/pMBNR2D (ura+), pMBNR1a/pMBNR2D/pMBNR2B (his+) or pMBNR1a/pMBNR2A/pMBNR2B, respectively, and was used to express the different NMDA receptor subunit genes.Western‐blotting analysis with the specific NMDA receptor antibodies showed a clear but differently strong expression of the recombinant receptor proteins which were found to be only partially glycosylated in the cell membranes of the recombinant yeast strains.By immunofluorescence microscopy using the specific subunit antibodies and fluorescence‐labeled secondary antibodies, the distinctly expressed NR1a and NR2D subunits could be located in the plasma membrane of the transformed yeast cells.Pharmacological characterization of crude membrane preparations of the recombinant yeast cells expressing 1−3 NMDA receptor subunits showed saturable binding of the glycine antagonist [3H]MDL105,519 with different Kd values of 56.88 ± 5.38 nM (NR1a), 1365.11 ± 76 nM (NR1a/NR2D), 22.97 ± 3.37 nM for NR1a/NR2B/NR2D and 7.4 ± 1.2 nM for NR1a/NR2A/NR2B. The bound capacities were 13.07 ± 0.92 (NR1a), 14.63 ± 0.50 (NR1a/NR2D), 12.85 ± 1.68 (NR1a/NR2B/NR2D) and 8.3 ± 0.7 (NR1a/NR2A/NR2B) pmol/mg membrane protein. The [3H]MDL105,519 binding was inhibited by the glycine antagonist 5,7‐dichlorokynurenate (DCKA), ethyl‐2‐carboxy‐4.6‐dichloro‐3‐indoleacetate (ECDI) and itself, but not by glycine,
D‐serine and 1‐amino‐cyclopropanecarboxylic acid (ACPC). Each of these recombinant receptor proteins consisting both of NR1 and NR2 subunits also showed a specific binding site for the NMDA agonist glutamate when using
L‐[3H]glutamate as a radioligand. Analysis of saturation experiments revealed that this ligand binds to a specific site with Kd values of 536 ± 43, 688 ± 60, and 856 ± 48 nM for NR1a/NR2B, NR1a/NR2D, and NR1a/NR2B/NR2D respectively.
Structural information on a cecropin‐like synthetic peptide, Shiva‐3 toxic to the sporogonic development of Plasmodium berghei
Authors: Boisbouvier Jérôme , Prochnicka‐Chalufour Ada , Nieto Alejandro R. , Torres Jorge A. , Nanard Nathalie , Rodriguez Mario H. , Possani Lourival D. , Delepierre Muriel 
Published: March 2003

This study is a contribution towards the understanding of the mode of action of Shiva‐3 and more generally that of cecropin‐like peptides. Structural information on Shiva‐3 (a cecropin‐like synthetic peptide) in water and in a membrane‐mimicking environment (trifluoroethanol alcohol, SDS) were obtained using analytical centrifugation, CD and NMR spectroscopies. A total of 20 converged structures were retained on the basis of 197 non‐redundant experimental constraints, including 166 distance constraints from the nuclear Overhauser effect measurements and 31 dihedral angle restraints derived from the purged COSY experiments. Some results obtained in presence of SDS are also presented.The toxic effects of the peptides obtained by cleavage (trypsin and lysine‐C hydrolysis) of Shiva‐3 on Escherichia coli and on Plasmodium berghei sporogonic stages are reported. Biological effects are discussed in relation to the calculated structure. The antiparasite activity and the low mosquito toxicity of Shiva‐3 make this peptide a good candidate for genetic transformation of mosquito vectors which warrants further studies aimed at the improvement of the molecule.
Structural determination of the O‐antigenic polysaccharide from Escherichia coli O35 and cross‐reactivity to Salmonella arizonae O62
Authors: Rundlöf Torgny , Weintraub Andrej , Widmalm Göran 
Published: March 2003

The structure of the O‐antigenic polysaccharide from Escherichia coli O35 has been investigated with the aid of NMR spectroscopy, sugar and methylation analyses. The sequence of the sugar residues could be determined by NOESY and heteronuclear‐multiple‐bond‐connectivity NMR experiments. The polysaccharide is composed of hexasaccharide repeating units with the following structure, where Rha and GalNAcAN represent rhamnose and 2‐acetamido‐2‐deoxy‐galacturonamide, respectively :

←3)‐β‐
D‐GlcpNAc‐(1←3)‐α‐
L‐Rhap‐(1←2)‐α‐
L‐Rhap‐(1←3)‐α‐
L‐Rhap‐(1←2)‐α‐
L‐Rhap‐(1←2↑1α‐ D‐GalpNAcANThe O‐antigen of Escherichia coli O35 is similar to the O‐specific polysaccharide from Salmonella arizonae O62, which instead has a terminal 2‐acetamido‐2‐deoxy‐α‐
D‐galacturonic acid residue. Immunochemical analyses using a rabbit antiserum specific for the Salmonella arizonae O62 O‐antigen showed an identical reactivity with both lipopolysaccharides.
Studies on the formation and stability of a complex between Streptomyces proteinaceous metalloprotease inhibitor and thermolysin
Authors: Kunugi Shigeru , Yanagi Yuichi , Oda Kohei 
Published: March 2003

The effects of certain physicochemical parameters on the formation and stability of a complex between Streptomyces proteinaceous metalloprotease inhibitor (SMPI) and thermolysin were investigated. SMPI had its lowest Ki value at a pH of around 6.5 (similar to the pH dependence of the kcat/Km of thermolysin catalysis), reflecting the splitting mechanism of the SMPI inhibition of thermolysin. This Ki increased with an increase in pressure, and ln (Ki–1) was almost linear with respect to pressure. The volume of the reaction (ΔVcomp), which is the volume change accompanying enzyme–inhibitor complex formation, was calculated as + 8.1 ± 0.3 mL·mol–1, which has a sign opposite to ΔVcomp for neutral peptide inhibitors and acyl‐peptide substrates. The temperature dependence of Ki–1 gave the reaction enthalpy (ΔHcomp) and reaction entropy (ΔScomp) of the complex formation as 34.6 ± 1.4 kJ·mol–1 and 298 ± 5 J·mol–1·K–1, respectively. These positive reaction volumes and reaction entropies were related to the electrostatic interactions and ionic strength dependence of Ki which corresponded to the key ionic interaction during complex formation. Complex formation with SMPI stabilized thermolysin against pressure perturbation as observed by the changes in the Trp fluorescence of thermolysin with increasing pressure. Thermal stability, however, was affected very little by complex formation with SMPI. Phosphoramidon, Cbz‐Phe‐Gly‐NH2 and Cbz‐Phe also positively affected the pressure‐tolerance of thermolysin, in the following order: Cbz‐Gly‐Phe‐NH2 < Cbz‐Phe < < phosphoramidon. The third compound exhibited stabilizing effects comparable with those of SMPI, which suggests that the interaction between SMPI and thermolysin was localized to the reactive site.
Substrate‐specific selenoprotein B of glycine reductase from  Eubacterium acidaminophilum
Authors: Wagner Matthias , Sonntag Denise , Grimm Rudolf , Pich Andreas , Eckerskorn Christoph , Söhling Brigitte , Andreesen Jan R. 
Published: March 2003

The substrate‐specific selenoprotein B of glycine reductase (PBglycine) from Eubacterium acidaminophilum was purified and characterized. The enzyme consisted of three different subunits with molecular masses of about 22 (α), 25 (β) and 47 kDa (γ), probably in an α2β2γ2 composition. PBglycine purified from cells grown in the presence of [75Se]selenite was labeled in the 47‐kDa subunit. The 22‐kDa and 47‐kDa subunits both reacted with fluorescein thiosemicarbazide, indicating the presence of a carbonyl compound. This carbonyl residue prevented N‐terminal sequencing of the 22‐kDa (α) subunit, but it could be removed for Edman degradation by incubation with o‐phenylenediamine. A DNA fragment was isolated and sequenced which encoded β and α subunits of PBglycine (grdE), followed by a gene encoding selenoprotein A (grdA2) and the γ subunit of PBglycine (grdB2). The cloned DNA fragment represented a second GrdB‐encoding gene slightly different from a previously identified partial grdB1‐containing fragment. Both grdB genes contained an in‐frame UGA codon which confirmed the observed selenium content of the 47‐kDa (γ) subunit. Peptide sequence analyses suggest that grdE encodes a proprotein which is cleaved into the previously sequenced N‐terminal 25‐kDa (β) subunit and a 22‐kDa (α) subunit of PBglycine. Cleavage most probably occurred at an ‐Asn‐Cys‐ site concomitantly with the generation of the blocking carbonyl moiety from cysteine at the α subunit.