Aldose reductase and the importance of experimental design

Kinetic studies on the AR (aldose reductase) protein have shown that it does not behave as a classical enzyme in relation to ring aldose sugars. As with non-enzymatic glycation reactions, there is probably a free radical element involved derived from monosaccharide autoxidation. in the case of AR, there is free radical oxidation of NADPH by autoxidizing monosaccharides, which is enhanced in the presence of the NADPH-binding protein. Thus any assay for AR based on the oxidation of NADPH in the presence of autoxidizing monosaccharides is invalid, and tissue AR measurements based on this method are also invalid, and should be reassessed. AR exhibits broad specificity for both hydrophilic and hydrophobic aldehydes that suggests that the protein may be involved in detoxification. The last thing we would want to do is to inhibit it. ARIs (AR inhibitors) have a number of actions in the cell which are not specific, and which do not involve them binding to AR. These include peroxy-radical scavenging and effects of metal ion chelation. The AR/ARI story emphasizes the importance of correct experimental design in all biocatalytic experiments. Developing the use of Bayesian utility functions, we have used a systematic method to identify the optimum experimental designs for a number of kinetic model data sets. This has led to the identification of trends between kinetic model types, sets of design rules and the key conclusion that such designs should be based on some prior knowledge of K-m and/or the kinetic model. We suggest an optimal and iterative method for selecting features of the design such as the substrate range, number of measurements and choice of intermediate points. The final design collects data suitable for accurate modelling and analysis and minimizes the error in the parameters estimated, and is suitable for simple or complex steady-state models.

Antixenosis to the peach/potato aphid, myzus persicae (SuIzer) in potato cultivars

Field studies were conducted to determine the antixenotic resistance against the peach-potato aphid, Myzus persicae (Sulzer) in different potato cultivars i.e. Cardinal, Ultimus and Desiree. These cultivars were studied in the paired tests as Cardinal vs. Desiree and Cardinal vs. Desiree for antixenosis (systemic insecticide-based test). In Desiree vs. Ultimus, no significant difference was found for the preference of M. persicae, while in case of Cardinal vs. Desiree, there was a significant preference for Desiree. Cardinal clearly showed antixenosis to M. persicae.

Reclassification of Bacteroides putredinis (Weinberg et al., 1937) in a New Genus Alistipes gen. nov., as Alistipes putredinis comb. nov., and description of Alistipes finegoldii sp nov., from human sources

During studies on the bacteriology of appendicitis in children, we often isolated from inflamed and non-inflamed tissue samples, an unusual bile-resistant pigment-producing strictly anaerobic gram-negative rod. Phenotypically this organism resembles members of Bacteroides fragilis group of species, as it is resistant to bile and exhibits a special-potency-disk pattern (resistance to vancomycin, kanamycin and colistin) typical for the B. fragilis group. However, the production of brown pigment on media containing haemolysed blood and a cellular fatty acid composition dominated by iso-C15:0, suggests that the organism most closely resembles species of the genus Porphyromonas. However, the unidentified organism differs from porphyromonads by being bile-resistant and by not producing butyrate as a metabolic end-product. Comparative 16S ribosomal RNA gene sequencing studies show the unidentified organism represents a distinct sub-line, associated with but distinct from, the miss-classified species Bacteroides putredinis. The clustering of the unidentified bacterium with Bacteroides putredinis was statistically significant, but they displayed >4% sequence divergence with each other. Chromosomal DNA-DNA pairing studies further confirmed the separateness of the unidentified bacterium and Bacteroides putredinis. Based on phenotypic and phylogenetic considerations, it is proposed that Bacteroides putredinis and the unidentified bacterium from human sources be classified in a new genus Alistipes, as Alistipes putredinis comb. nov. and Alistipes finegoldii sp. nov., respectively. The type strain of Alistipes finegoldii is CCUG 46020(T) (= AHN2437(T)).

Determination of digesta flow entering the omasal canal of dairy cows using different marker systems

Four studies were conducted to compare the effect of four indigestible markers (LiCoEDTA, Yb-acetate, Cr-mordanted straw and indigestible neutral-detergent fibre (INDF)) and three marker systems on the flow of digesta entering the omasal canal of lactating dairy cows. Samples of digesta aspirated from the omasal canal were pooled and separated using filtration and high-speed centrifugation into three fractions defined as the liquid phase, small particulate and large particulate matter. Co was primarily associated with the liquid phase, Yb was concentrated in small particulate matter, whilst Cr and INDF were associated with large particles. Digesta flow was calculated based on single markers or using the reconstitution system based on combinations of two (Co + Yb, Co + Cr and Co + INDF) or three markers (Co + Yb + Cr and Co + Yb + INDF). Use of single markers resulted in large differences between estimates of organic matter (OM) flow entering the omasal canal suggesting that samples were not representative of true digesta. Digesta appeared to consist of at least three phases that tended to separate during sampling. OM was concentrated in particulate matter, whilst the liquid phase consisted mainly of volatile fatty acids and inorganic matter. Yb was intimately associated with nitrogenous compounds, whereas Cr and INDF were concentrated in fibrous material. Current data indicated that marker systems based on Yb in combination with Cr or INDF are required for the accurate determination of OM, N and neutral-detergent fibre flow. In cases where the flow of water-soluble nutrients entering the omasal canal is also required, the marker system should also include Co.

Posttraumatic stress disorder following assault: the role of cognitive processing, trauma memory, and appraisals

Two studies of assault victims examined the roles of (a) disorganized trauma memories in the development of posttraumatic stress disorder (PTSD), (b) peritraumatic cognitive processing in the development of problematic memories and PTSD, and (c) ongoing dissociation and negative appraisals of memories in maintaining symptomatology. In the cross-sectional study (n = 81), comparisons of current, past, and no-PTSD groups suggested that peritraumatic cognitive processing is related to the development of disorganized memories and PTSD. Ongoing dissociation and negative appraisals served to maintain PTSD symptoms. The prospective study (n = 73) replicated these findings longitudinally. Cognitive and memory assessments completed within 12-weeks postassault predicted 6-month symptoms. Assault severity measures explained 22% of symptom variance; measures of cognitive processing, memory disorganization, and appraisals increased prediction accuracy to 71%.

‘Trying to be a Man': J. M. Barrie and sentimental masculinity

This article discusses the treatment of sentimentality in the fiction of J.M. Barrie, focusing in particular on Tommy and Grizel (1900). It place the discussion in the context of wider debates over sentimentality in Victorian culture and explores the intersection between these and discourse of gender and sexuality in the late nineteenth century.

To 'be', and not to 'have': auxiliary selection in unaccusative verbs in Italian

This paper investigates the characteristics of unaccusative verbs in Italian with respect to the consistency with which these verbs select the auxiliaries ‘be’ (essere) and ‘have’ (avere) in compound tense forms. The study builds on the gradient approach to split intransitivity (Sorace 2000) by exploring the behaviour of 29 intransitive Italian verbs with respect to their core-peripheral features: auxiliary selection acceptability ratings and associated variance measures. Although there is clear support for the gradient approach in relation to the general order of semantic categories along the unaccusativity gradient, the results reveal that the ordering of subclasses within the Change group conflict with that currently proposed in the literature. In addition, the findings demonstrate the aspectual and lexical semantic characteristics of internally-caused change-of-state verbs in Italian require further investigation before their auxiliary selection behaviour can be properly understood. Furthermore, contrary to the gradient account, Existence verbs, the most stative and therefore the most peripheral subclass in the unaccusativity hierarchy, exhibit behaviour more characteristic of core unaccusative verbs. This study examines a wider range of semantic subclasses of unaccusative verbs than has hitherto been reported and identifies the core-peripheral boundary for Italian.1

‘Souvenirs, souvenirs': the role of memory in the work of Annie Saumont

The work of nouvelliste Annie Saumont constantly explores the phenomenon of memory, and of memories. This article identifies and nuances the various forms that this exploration takes. An introductory contextualization of author and theme is followed by the presentation of a short story, ‘Méandres’, which embodies the first quality of memory to be examined: its capacity not only to recall but also to re-evaluate a past which is thus shown to be as hypothetical as the future. Memory as guilt that moulds or puts its indelible stamp on lives is then evoked by means of examples from other stories, illustrating the gradations Saumont achieves in her investigation of the power of this complex faculty. The next section turns to her portrayal of involuntary memory. Unlike for Proust, the instances of spontaneous remembering that are experienced by her characters lunge at them down the years almost exclusively to wound or disorientate. Depictions of the memory which conserves, and is thus burdened by, secrets are then considered, and finally Saumont's evocation of characters who have different reasons to analyse the way their own and other people's memories work. The conclusion to be drawn is that for Saumont, we are our memories; the ability to master a ‘judicious interpretation’ of memory – or indeed, to forget – is, in her stories, overwhelmingly a quality to be envied.

Membrane ruffling and invasion of human and avian cell lines is reduced for aflagellate mutants of Salmonella enterica serotype Enteritidis

Independent studies have demonstrated that flagella are associated with the invasive process of Salmonella enterica serotypes, and aflagellate derivatives of Salmonella enterica serotype Enteritidis are attenuated in murine and avian models of infection. One widely held view is that the motility afforded by flagella, probably aided by chemotactic responses, mediates the initial interaction between bacterium and host cell. The adherence and invasion properties of two S. Enteritidis wild-type strains and isogenic aflagellate mutants were assessed on HEp-2 and Div-1 cells that are of human and avian epithelial origin, respectively. Both aflagellate derivatives showed a significant reduction of invasion compared with wild type over the three hours of the assays. Complementation of the defective fliC allele recovered partially the wild-type phenotype. Examination of the bacterium-host cell interaction by electron and confocal microscopy approaches showed that wild-type bacteria induced ruffle formation and significant cytoskeletal rearrangements on HEp-2 cells within 5 minutes of contact. The aflagellate derivatives induced fewer ruffles than wild type. Ruffle formation on the Div-1 cell line was less pronounced than for HEp-2 cells for wild-type S. Enteritidis. Collectively, these data support the hypothesis that flagella play an active role in the early events of the invasive process.

The acquisition of person and number morphology within the verbal domain in Early Greek

The aim of this paper is to examine the acquisition pattern of person and number verb morphology within the generative framework and to compare the results of the analyses with previous research in Greek and other European languages. The study considers previous data on the acquisition of subject-verb agreement, and thereafter, examines the acquisition of person and number morphology in a new dataset of two monolingual Greek-speaking children. The analyses present quantitative data of accuracy of person and number marking, error data, and qualitative analyses addressing the productivity of person and number marking. The results suggest that person and number morphology is used correctly and productively from a very early age in Greek speaking children. The findings provide new insight into early Greek language acquisition and are also relevant for research in early development of languages with rich inflectional morphology.