bluuebunny/crossref_metadata_2025_split huggingface dataset:
Is ‘shared decision‐making’ feasible in consultations for upper respiratory tract infections? Assessing the influence of antibiotic expectations using discourse analysis
Authors: Elwyn Glyn , Gwyn Richard , Edwards Adrian , Grol Richard 
Published: March 2003

ObjectivesTo examine the discourse of consultations in which conflict occurs between parents and clinicians about the necessity of antibiotics to treat an upper respiratory tract infection. To appraise the feasibility of shared decision‐making in such consultations.DesignA qualitative study using discourse analysis techniques.SettingA general practice with 12 500 patients in an urban area of Cardiff, Wales.ParticipantsTwo consultations were purposively selected from a number of audiotaped sessions. The consultations took place during normal clinics in which appointments are booked at 7‐minute intervals. The practitioner is known to be interested in involving patients in treatment decisions.MethodDiscourse analysis was employed to examine the consultation transcripts. This analysis was then compared with the theoretical competencies proposed for ‘shared decision‐making’.ResultsThe consultations exhibit less rational strategies than those suggested by the shared decision‐making model. Strong parental views are expressed (overtly and covertly) which seem derived from prior experiences of similar illnesses and prescribing behaviours. The clinician responds by emphasizing the ‘normality’ of upper respiratory tract infections and their recurrence, accompanied by expressions that antibiotic treatment is ineffective in ‘viral’ illness – the suggested diagnosis. The competencies of ‘shared decision‐making’ are not exhibited.ConclusionsThe current understanding of shared decision‐making needs to be developed for those situations where there are dis‐agreements due to the strongly held views of the participants. Clinicians have limited strategies in situations where patient treatment preferences are opposed to professional views. Dispelling ‘misconceptions’ by sharing information and negotiating agreed management plans are recommended. But it seems that communication skills, information content and consultation length have to receive attention if such strategies are to be employed successfully.
A qualitative study of women’s views about how health professionals communicate about infant feeding
Authors: Hoddinott Pat , Pill Roisin 
Published: March 2003

ObjectiveTo look at how communication by health professionals about infant feeding is perceived by first time mothers.DesignQualitative semi‐structured interviews early in pregnancy and 6–10 weeks after birth.Subjects and settingTwenty‐one white, low income women expecting their first baby were interviewed mostly at home, often with their partner or a relative.ResultsThe personal and practical aspects of infant feeding which were important to women were seldom discussed in detail in ante‐natal interviews. In post‐natal interviews women described how words alone encouraging them to breastfeed were insufficient. Apprenticeship style learning of practical skills was valued, particularly time patiently spent watching them feed their baby. Women preferred to be shown skills rather than be told how to do them. Some felt pressure to breastfeed and bottle feeding mothers on post‐natal wards felt neglected in comparison. Women preferred their own decision‐making to be facilitated rather than being advised what to do. Some women experienced distress exposing their breasts and being touched by health professionals. Continuity of care and forming a personal relationship with a health professional who could reassure them were key factors associated with satisfaction with infant feeding communication.ConclusionsThe infant feeding goal for many women is a contented, thriving baby. In contrast, women perceive that the goal for health professionals is the continuation of breastfeeding. These differing goals can give rise to dissatisfaction with communication which is often seen as ‘breastfeeding centred’ rather than ‘woman centred.’ Words alone offering support for breastfeeding were often inadequate and women valued practical demonstrations and being shown how to feed their baby. Spending time with a caring midwife with whom the woman had developed a personal, continuing relationship was highly valued. Women were keen to maintain ownership, control and responsibility for their own decision‐making about infant feeding.
Development and implementation of a science training course for breast cancer activists: Project LEAD (leadership, education and advocacy development)
Authors: Dickersin Kay , Braun Lundy , Mead Margaret , Millikan Robert , Wu Anna M. , Pietenpol Jennifer , Troyan Susan , Anderson Benjamin , Visco Frances 
Published: March 2003

Objective To develop and implement Project LEAD (leadership, education, and advocacy development), a science course for breast cancer activists. Population Students were breast cancer activists and other consumers, mainly affiliated with advocacy organizations in the United States of America. Setting Project LEAD is offered by the National Breast Cancer Coalition; the course takes place over 5 days and is offered 4 times a year, in various cities in the United States of America. Results The Project LEAD curriculum has developed over 5 years to include lectures, problem‐based study groups, case studies, interactive critical appraisal sessions, a seminar by an ‘expert’ scientist, role play, and homework components. A core faculty has been valuable for evaluating and revising the course and has proved necessary to provide consistent high quality teaching. Course evaluations indicated that students gained critical appraisal skills, enhanced their knowledge and developed confidence in selected areas of basic science and epidemiology. Conclusions Project LEAD comprises a unique curriculum for training breast cancer activists in science and critical appraisal. Course evaluations indicate that students gain confidence and skills from the course.
Synthesis of phosphoenol pyruvate (PEP) analogues and evaluation as inhibitors of PEP‐utilizing enzymes
Authors: García‐Alles Luis Fernando , Erni Bernhard 
Published: March 2003

The synthesis of 10 new phosphoenolpyruvate (PEP) analogues with modifications in the phosphate and the carboxylate function is described. Included are two potential irreversible inhibitors of PEP‐utilizing enzymes. One incorporates a reactive chloromethylphosphonate function replacing the phosphate group of PEP. The second contains a chloromethyl group substituting for the carboxylate function of PEP. An improved procedure for the preparation of the known (Z)‐ and (E)‐3‐chloro‐PEP is also given. The isomers were obtained as a 4 : 1 mixture, resolved by anion‐exchange chromatography after the last reaction step. The stereochemistry of the two isomers was unequivocally assigned from the 3JH‐C coupling constants between the carboxylate carbons and the vinyl protons.All of these and other known PEP‐analogues were tested as reversible and irreversible inhibitors of Mg2+‐ and Mn2+‐ activated PEP‐utilizing enzymes: enzyme I of the phosphoenolpyruvate:sugar phosphotransferase system (PTS), pyruvate kinase, PEP carboxylase and enolase. Without exception, the most potent inhibitors were those with substitution of a vinyl proton. Modification of the phosphate and the carboxylate groups resulted in less effective compounds. Enzyme I was the least tolerant to such modifications. Among the carboxylate‐modified analogues, only those replaced by a negatively charged group inhibited pyruvate kinase and enolase. Remarkably, the activity of PEP carboxylase was stimulated by derivatives with neutral groups at this position in the presence of Mg2+, but not with Mn2+. For the irreversible inhibition of these enzymes, (Z)‐3‐Cl‐PEP was found to be a very fast‐acting and efficient suicide inhibitor of enzyme I (t1/2 = 0.7 min).
Structural analysis of deacylated lipopolysaccharide of Escherichia coli strains 2513 (R4 core‐type) and F653 (R3 core‐type)
Authors: Müller‐Loennies Sven , Lindner Buko , Brade Helmut 
Published: March 2003

Lipopolysaccharide (LPS) of Escherichia coli strain 2513 (R4 core‐type) yielded after alkaline deacylation one major oligosaccharide by high‐performance anion‐exchange chromatography (HPAEC) which had a molecular mass of 2486.59 Da as determined by electrospray ionization mass spectrometry. This was in accordance with the calculated molecular mass of a tetraphosphorylated dodecasaccharide of the composition shown below. NMR‐analyses identified the chemical structure aswhere l‐α‐d‐Hep is l‐glycero‐α‐d‐manno‐heptopyranose and Kdo is 3‐deoxy‐α‐d‐manno‐oct‐2‐ulopyranosylonic acid and all hexoses are present as d‐pyranoses.We have also isolated the complete core‐oligosaccharides of E. coli F653 LPS for which only preliminary data were available and investigated the deacylated LPS by NMR and MS. The proposed structure determined previously by methylation analysis was confirmed and is shown below.In addition we have quantified the side‐chain heptose substitution of the inner core with GlcpN (≈ 30%) and confirmed that this sugar is only present when the phosphate at the second l,d‐Hepp residue is absent.
Thermoanaerobacterium thermosulfurigenes cyclodextrin glycosyltransferase
Authors: Leemhuis Hans , Dijkstra Bauke W. , Dijkhuizen Lubbert 
Published: March 2003

Cyclodextrin glycosyltransferase (CGTase) uses an α‐retaining double displacement mechanism to catalyze three distinct transglycosylation reactions. To investigate these reactions as catalyzed by the CGTase from Thermoanaerobacterium thermosulfurigenes the enzyme was overproduced (8 mg·L−1 culture) using Bacillus subtilis as a host. Detailed analysis revealed that the three reactions proceed via different kinetic mechanisms. The cyclization reaction (cyclodextrin formation from starch) is a one‐substrate reaction, whereas the other two transglycosylation reactions are two‐substrate reactions, which obey substituted enzyme mechanism kinetics (disproportionation reaction) or ternary complex mechanism kinetics (coupling reaction).Analysis of the effects of acarbose and cyclodextrins on the disproportionation reaction revealed that cyclodextrins are competitive inhibitors, whereas acarbose is a mixed type of inhibitor. Our results show that one molecule of acarbose binds either in the active site of the free enzyme, or at a secondary site of the enzyme–substrate complex. The mixed inhibition thus indicates the existence of a secondary sugar binding site near the active site of T. thermosulfurigenes CGTase.
Structural and functional divergence of class II alcohol dehydrogenase
Authors: Svensson Stefan , Hedberg Jesper J. , Höög Jan‐Olov 
Published: March 2003

cDNAs coding for class II alcohol dehydrogenase were isolated from a rabbit‐liver cDNA library. Deduced amino acid sequences show that isozymic forms of rabbit class II alcohol dehydrogenase exist, with a positional identity of 88.4 %. A high variability in structure of class II alcohol dehydrogenase between the species is also reflected in function. The rabbit II‐1 isozyme shows common characteristics with the human enzyme, but has a lower Km value for ethanol, 4.2 mM. The II‐2 isozyme shows restriction for aliphatic alcohols longer than pentanol. For shorter alcohols the II‐2 form has similar Km values as the II‐1 isozyme, 5.5 mM for ethanol, but is a low activity variant with a 10‐fold decrease in kcat values compared with II‐1. Nevertheless, II‐2 has a higher specificity for benzoquinone than II‐1 due to a lower Km value, 80 μM compared with 1 mM, and is in this sense more like the human class II enzyme. In addition a rabbit class III alcohol dehydrogenase cDNA was isolated that encodes a typical class III enzyme/glutathione‐dependent formaldehyde dehydrogenase.The finding of isozymic forms of class II alcohol dehydrogenase is in line with the evolution of the system of medium‐chain alcohol dehydrogenases with different enzymes, different classes and different isozymes and further underline the complexity of the entire mammalian alcohol dehydrogenase system.
The mechanism of porcine pancreatic α‐amylase
Authors: Al Kazaz Modhafar , Desseaux Véronique , Marchis‐Mouren Guy , Prodanov Eugenio , Santimone Marius 
Published: March 2003

A kinetic study was carried out on the inhibitory effects of acarbose, maltose, and maltotriose on porcine pancreatic α‐amylase (PPA), using maltopentaose as the substrate.Lineweaver‐Burk plots showed that the inhibitory action of acarbose is of the mixed non‐competitive type. The secondary plots gave straight lines. A model involving abortive complexes accounts for these results. Dixon plot analysis led to the same conclusion. According to the proposed model, one molecule of acarbose per amylase molecule binds either directly to free enzyme at the active site or to the enzyme‐substrate complex at a secondary carbohydrate‐binding site, which becomes functional after the substrate has bound to the enzyme molecule at the active site.Kinetic analysis of the inhibition exerted by either the maltose or maltotriose reaction products of maltopentaose hydrolysis were then performed. The inhibitory effect of maltose was found to be of the non‐competitive type, while that of maltotriose was competitive. It can therefore be concluded that the first reaction product to be released upon maltopentaose hydrolysis is maltose, and that the second product is maltotriose. This indicates that after hydrolysis of the maltopentaose chain, the reducing side fragment is released first.
Labeling of Torpedo californica nicotinic acetylcholine receptor subunits by cobratoxin derivatives with photoactivatable groups of different chemical nature at Lys23
Authors: Utkin Yuri N. , Krivoshein Arcadius V. , Davydov Vasily L. , Kasheverov Igor E. , Franke Peter , Maslennikov Innokenty V. , Arseniev Alexander S. , Hucho Ferdinand , Tsetlin Victor I. 
Published: March 2003

Different photoactivatable derivatives of toxin 3 (CTX) Naja naja siamensis were obtained after CTX reaction with N‐hydroxysuccinimide esters of p‐azidobenzoic, p‐azidotetraflourobenzoic, p‐benzoylbenzoic and p‐[3‐(trifluoromethyl)‐3H‐diazirin‐3‐yl]benzoic acids. The ion‐exchange HPLC profiles for the reaction products were very similar in four cases, with one predominant peak corresponding to the derivative containing the label at Lys23. After [125I]iodination, CTX photoactivatable derivatives were cross‐linked to the nicotinic acetylcholine receptor from Torpedo californica under optimized conditions. The highest cross‐linking yield (up to 16 % of the bound toxin) was observed for azidobenzoyl‐Lys23‐CTX. Different receptor subunits were found to be labelled depending on the nature of the photoactivatable group : the azido derivatives labelled the γ and δ subunits, benzoylbenzoyl derivative labelled the α and δ subunits, while p‐[3‐(trifluoromethyl)‐3H‐diazirin‐3‐yl]benzoyl derivative reacted with α, γ and δ subunits.The cross‐linking experiments in the presence of varying concentrations of (+)‐tubocurarine demonstrated that the Lys23‐attached diazirinyl group contacts the δ and α subunits in one ligand‐binding site, whereas at the other site, for another CTX molecule, the contacts of the Lys23‐diazirinyl are with γ and α subunits. This means that the central loop in the two CTX molecules binds at the α/γ and α/δ interfaces. Calculation of the sterically possible displacement of diazirinyl nitrogen, basing on the known X‐ray structure of CTX, showed that this value does not exceed 13 Å. The results obtained favor the disposition of the ligand‐binding sites at the subunit interfaces, with the distance between α and δ, or α and γ subunits at these sites being not more than 13 Å.
Oligonucleotide facilitators enable a hammerhead ribozyme to cleave long RNA substrates with multiple‐turnover activity
Authors: Jankowsky Eckhard , Schwenzer Bernd 
Published: March 2003

Trans‐acting hammerhead ribozymes are usually efficient in cleaving short RNA model substrates under both single‐turnover and multiple‐turnover conditions. In contrast, when long RNAs are the substrates, the cleavage efficiency of these ribozymes decreases, including a loss of multiple‐turnover activity in many cases. Since target substrates for potential therapeutical purposes are mostly long RNAs, a multiple‐turnover cleavage of long RNAs would essentially increase the efficiency of hammerhead ribozymes. Therefore, we explored if oligonucleotide facilitators, capable of enhancing multiple‐turnover activity with short substrates, can also affect or cause multiple turnover with long substrates. We examined the effects of 12‐base and 24‐base oligonucleotide facilitators on the multiple‐turnover activity with substrates of different length containing 39‐, 452‐ and 942‐base sequences of the human tissue factor (HTF) mRNA. In the absence of facilitator, the ribozyme cleaved only the 39‐base substrate with multiple‐turnover activity, but not the long 452‐base and 942‐base substrates. However, facilitator addition enabled the ribozyme to cleave even the 452‐base and the 942‐base substrates with multiple‐turnover activity. All facilitators tested showed a remarkable activating effect with the long substrates.The data demonstrate that a hammerhead ribozyme which, by itself, can only act as a single‐turnover catalyst with long substrates, can be switched by facilitators into a multiple‐turnover catalyst. Thus, the inactivation of long target RNAs in multiple‐turnover reactions may be achieved by addition of oligonucleotide facilitators.
Calcitriol and transforming growth factor‐β upregulate 5‐lipoxygenase mRNA expression by increasing gene transcription and mRNA maturation
Authors: Härle Damaris , Rådmark Olof , Samuelsson Bengt , Steinhilber Dieter 
Published: March 2003

Differentiation of the human monocytic cell line Mono Mac 6 with calcitriol plus transforming growth factor‐β (TGFβ) strongly induces 5‐lipoxygenase (5‐LO) mRNA and protein expression. The mechanism of 5‐LO mRNA induction by these agents was investigated. Analysis of mature 5‐LO mRNA by reverse‐transcriptase‐PCR gave a 42‐fold induction which was not due to alterations in 5‐LO half life and which was only in part due to an induction of gene transcription. There was an up to fivefold increase in 5‐LO primary transcripts by TGFβ and calcitriol, which could be inhibited by cycloheximide. No significant effects on 5‐LO transcription were observed with TGFβ or calcitriol alone. However, treatment of the cells with either calcitriol or TGFβ and addition of the corresponding second inducer lead to an about fourfold induction of primary transcript levels. Addition of cycloheximide together with the second inducer inhibited only the TGFβ but not the calcitriol effects, which indicated that there is a direct stimulation of 5‐LO transcription by calcitriol in the presence of TGFβ‐induced proteins.In order to investigate the effects of TGFβ/calcitriol on 5‐LO transcript, elongation and maturation, the relative changes in immature and mature 5‐LO RNA species were analyzed by reverse‐transcription‐PCR. Analysis of exons 1−5 indicated an about threefold induction of 5‐LO transcripts by calcitriol/TGFβ, respectively. However, when exons 6−14 were determined, more pronounced increments were found (3.6−12‐fold). Selective analysis of polyadenylated and spliced 5‐LO mRNA species gave a 42‐fold induction. The effects of both TGFβ and calcitriol on transcript elongation and maturation were inhibited by cycloheximide.Our results show that induction of 5‐LO mRNA by calcitriol and TGFβ is due to a modest increase in 5‐LO gene transcription and to the stimulation of transcript elongation and maturation.
Purification and characterization of an alcohol dehydrogenase from the Antarctic psychrophile Moraxella sp. TAE123
Authors: Tsigos Iason , Velonia Kelly , Smonou Ioulia , Bouriotis Vassilis 
Published: March 2003

An NAD+‐dependent alcohol dehydrogenase (ADH) of the Antarctic psychrophile Moraxella sp. TAE123 was purified to homogeneity with an overall yield of 16.7  % and further characterized. The native enzyme had an apparent molecular mass of 240 kDa and consisted of four identical 52‐kDa subunits. The pI of the enzyme was determined to be 5.5, while its optimum pH is 7.5. The enzyme contained 1 zinc atom/subunit and exhibited a remarkable thermal lability. Moraxella sp. TAE123 ADH exhibited a wide range of substrate specificity similar to its mammalian counterparts and in contrast to other microbial ADHs. It oxidized mainly primary and secondary aliphatic alcohols. The highest reaction rate was observed when ethanol was used as substrate. A gradual decrease in rate was observed by increasing the length and branching of the carbon chain of the alcohol. This enzyme oxidized effectively large bulky alcohols, such as diphenylmethanol. Reduction of aldehydes and ketones was also observed.N‐terminal amino acid sequence analysis of the enzyme did not reveal any similarity with the amino termini of all other ADHs, while an unexpected significant similarity was observed with the amino terminal sequence of four prokaryotic aldehyde dehydrogenases.
Characterization of three transcriptional repressor sites within the 3′ untranslated region of the rat serine protease inhibitor 2.3 gene
Authors: Paul Conception , Simar‐Blanchet Anne‐Emmanuelle , Ro Hyo‐Sung , Le Cam Alphonse 
Published: March 2003

The activity of the rat serine protease inhibitor 2.3 gene (spi 2.3) is controlled by several positive promoter elements [Simar‐Blanchet, A.‐E., Paul, C., Mercier, L. & Le Cam, A. (1996) Eur. J. Biochem. 236, 638−648] and a negative element located in the 3′ untranslated gene region (3′ UTR) [Le Cam, A. & Legraverend, C. (1996) Eur. J. Biochem. 231, 620−627]. In the present studies, we dissected the 348‐bp spi 2.3 3′ UTR silencer to precisely define repressor sites and look for specifically interacting proteins. Three short elements referred to as A (nucleotides 1751−1776 in the cDNA), B (nucleotides 1812−1827) and C (nucleotides 1958−1974) sites repressed transcription from the homologous spi 2.3 promoter as well as from a heterologous minimal promoter containing the spi GAGA box enhancer. All three sites harbor a (TTTC) motif whose mutation affected silencer activity that was also dependent on flanking sequences. Those sites share the (TTTC) motif and a CCAAT/enhancer‐binding‐protein(C/EBP)−binding site with a fatty‐acid‐binding‐protein gene promoter element shown to interact specifically with a transcriptional repressor [He, G. P., Muise, A., Wu Li, A. & Ro, H.‐S. (1995) Nature 378, 92−96]. This repressor is however unlikely to mediate spi 2.3 3′ UTR silencer action since it was not detected in rat hepatocytes. In vitro footprinting of the spi 2.3 3′ UTR silencer region revealed a strong interaction with liver nuclear proteins. Among the six identified footprints, three of them (F‐II, FIII and F‐IV) bound C/EBPs and mapped in regions harboring the repressor function. Binding of C/EBPs to all three spi 2.3 3′ UTR repressor sites, although rather weak, was confirmed by electrophoretic mobility shift assays that otherwise failed to reveal specific interactions with other liver nuclear proteins in vitro. However, none of the most largely liver expressed C/EBP species (i.e. α, β and δ) activated the spi 2.3 3′ UTR silencer function in NIH 3T3 cells, suggesting that binding of those transcription factors did not mediate the transcriptional repression.