Isolation of AF2 (KHEYLRFamide) from Caenorhabditis elegans: Evidence for the presence of more than one FMRFamide related peptide-encoding gene

Numerous FMRF amide-related peptides (FaRPs) have been isolated and sequenced from extracts of free-living and parasitic nematodes. The most abundant FaRP identified in ethanolic/methanolic extracts of the parasitic forms, Ascaris suum and Haemonchus contortus and from the free-living nematode, Panagrellus redivivus, was KHEYLRF amide (AF2). Analysis of the nucleotide sequences of cloned FaRP-precursor genes from C. elegans and, more recently, Caenorhabditis vulgaris identified a series of related FaRPs which did not include AF2. An acid-ethanol extract of Caenorhabditis elegans was screened radioimmunometrically for the presence of FaRPs using a C-terminally directed FaRP antiserum. Approximately 300 pmols of the most abundant immunoreactive peptide was purified to homogeneity and 30 pmols was subjected to Edman degradation analysis and gas-phase sequencing. The unequivocal primary structure of the heptapeptide, Lys-His-Glu-Tyr-Leu-Arg-Phe-NH2 (AF2) was determined following a single gas-phase sequencing run. The molecular mass of the peptide was determined using a time-of-flight mass spectrometer and was found to be 920 (MH(+))(-), which was consistent with the theoretical mass of C-terminally amidated AF2. These results indicate that C. elegans possesses more than one FaRP gene. (C) 1995 Academic Press, Inc.

Characterization of the highly conserved VFMGD motif in a bacterial polyisoprenyl-phosphate N-acetylaminosugar-1-phosphate transferase

Polyisoprenyl-phosphate N-acetylaminosugar-1-phosphate transferases (PNPTs) constitute a family of eukaryotic and prokaryotic membrane proteins that catalyze the transfer of a sugar-1-phosphate to a phosphoisoprenyl lipid carrier. All PNPT members share a highly conserved 213-Valine-Phenylalanine-Methionine-Glycine-Aspartic acid-217 (VFMGD) motif. Previous studies using the MraY protein suggested that the aspartic acid residue in this motif, D267, is a nucleophile for a proposed double-displacement mechanism involving the cleavage of the phosphoanhydride bond of the nucleoside. Here, we demonstrate that the corresponding residue in the E. coli WecA, D217, is not directly involved in catalysis, as its replacement by asparagine results in a more active enzyme. Kinetic data indicate that the D217N replacement leads to more than twofold increase in V(max) without significant change in the K(m) for the nucleoside sugar substrate. Furthermore, no differences in the binding of the reaction intermediate analog tunicamycin were found in D217N as well as in other replacement mutants at the same position. We also found that alanine substitutions in various residues of the VFMGD motif affect to various degrees the enzymatic activity of WecA in vivo and in vitro. Together, our data suggest that the highly conserved VFMGD motif defines a common region in PNPT proteins that contributes to the active site and is likely involved in the release of the reaction product.

A calcium-dependent interaction between calmodulin and the calponin homology domain of human IQGAP1

IQGAPs are cytoskeletal scaffolding proteins which collect information from a variety of signalling pathways and pass it on to the microfilaments and microtubules. There is a well-characterised interaction between IQGAP and calmodulin through a series of IQ-motifs towards the middle of the primary sequence. However, it has been shown previously that the calponin homology domain (CHD), located at the N-terminus of the protein, can also interact weakly with calmodulin. Using a recombinant fragment of human IQGAP1 which encompasses the CHD, we have demonstrated that the CHD undergoes a calcium ion-dependent interaction with calmodulin. The CHD can also displace the hydrophobic fluorescent probe 1-anilinonaphthalene-8-sulphonate from calcium-calmodulin, suggesting that the interaction involves non-polar residues on the surface of calmodulin. Molecular modelling identified a possible site on the CHD for calmodulin interaction. The physiological significance of this interaction remains to be discovered.

Understanding Racial Tensions in Northern Ireland: Threat, Paranoia and the Fear of Loss

This paper will explore the development of increased group tensions in Northern Ireland
over the past decade with a special emphasis being placed upon rising racial tensions in cities such
as Belfast and Lisburn. The paper will analyse why Northern Ireland has been described as the new
race-hate capital of Europe and, through a case-study of Loyalism, will argue that if this growth in
racist sentiment is to be prevented, more needs to be done to understand the causes of such feeling,
particularly within loyalist working-class areas. I will argue that society as a whole needs to address
the fears and anxieties of those that perceive themselves to be under threat from the recent increase
in immigration or else we risk creating a new cause célèbre for those that would seek to extend the
lifetime of our paramilitary organisations. Moreover, at a time when loyalist communities feel politically
alienated and lacking representation, there is a real danger of British far-right groups exploiting the
situation and making long-term political capital.

Aminopeptidases of malaria parasites:new targets for chemotherapy

Novel targets for new drug development are urgently required to combat malaria, a disease that puts half of the world's population at risk. One group of enzymes identified within the genome of the most lethal of the causative agents of malaria, Plasmodium falciparum, that may have the potential to become new targets for antimalarial drug development are the aminopeptidases. These enzymes catalyse the cleavage of the N-terminal amino acids from proteins and peptides. P. falciparum appears to encode for at least nine aminopeptidases, two neutral aminopeptidases, one aspartyl aminopeptidase, one aminopeptidase P, one prolyl aminopeptidase and four methionine aminopeptidases. Recent advances in our understanding of these genes and their protein products are outlined in this review, including their potential for antimalarial drug development.

Constraint rule-based programming of norms for electronic institutions

Norms constitute a powerful coordination mechanism among heterogeneous agents. In this paper, we propose a rule language to specify and explicitly manage the normative positions of agents (permissions, prohibitions and obligations), with which distinct deontic notions and their relationships can be captured. Our rule-based formalism includes constraints for more expressiveness and precision and allows to supplement (and implement) electronic institutions with norms. We also show how some normative aspects are given computational interpretation. © 2008 Springer Science+Business Media, LLC.

Hypercyclic operators on countably dimensional spaces

According to Grivaux, the group GL(X) of invertible linear operators on a separable infinite dimensional Banach space X acts transitively on the set s (X) of countable dense linearly independent subsets of X. As a consequence, each A? s (X) is an orbit of a hypercyclic operator on X. Furthermore, every countably dimensional normed space supports a hypercyclic operator. Recently Albanese extended this result to Fréchet spaces supporting a continuous norm. We show that for a separable infinite dimensional Fréchet space X, GL(X) acts transitively on s (X) if and only if X possesses a continuous norm. We also prove that every countably dimensional metrizable locally convex space supports a hypercyclic operator.

The systematic development of an innovative DVD to raise awareness of preconception care

The purpose of this article is to describe the design, development and process evaluation of a preconception counselling resource (a DVD) for women with pre-gestational diabetes. DVD design and development centred on two key stakeholders ('DVD user group' and 'professional advisory group') working alongside a professional multimedia company. The DVD user group provided feedback on preferred DVD style, and informed modifications and improvements. The professional advisory group prepared the script, and ensured content and face validity. Evaluation of the DVD's acceptability and usefulness was assessed among women with diabetes via a postal questionnaire. Development phase: the resulting DVD is a 45-minute programme with three parts, featuring eight women with diabetes sharing their views and experiences, alongside an evidence-based commentary. The programme focuses on the importance of preventing an unplanned pregnancy (highlighting contraception) and on essential planning advice. Evaluation phase: 97 women (89 with type 1 and 8 with type 2 diabetes) evaluated the DVD using a rating scale of 0-10. Mean (SD) scores were: 9.1 (1.3) for quality; 9.0 (1.4) for content; 8.8 (1.5) for interest; 8.7 (1.8) for usefulness; 7.8 (2.2) for knowledge acquisition; and 8.0 (2.1) for knowledge confirmation. This combined user and multi-professional advisory group approach has produced an innovative and highly acceptable preconception counselling resource for women with diabetes. The development process and outcome evaluation are an important point of reference for future educational programmes. Future research will evaluate the impact of this preconception counselling resource on pregnancy planning indicators and pregnancy outcome.

Autobiographical memory specificity, depression, and trauma in bipolar disorder

Objectives. In a bipolar disorder (BD) sample, the present study investigated: (i) the prevalence of trauma; (ii) the specificity of autobiographical memory (AM); (iii) the influence of childhood trauma on AM specificity, current inter-episode depressive mood, and BD severity; (iv) if AM specificity moderates the influence of childhood trauma on current inter-episode depressive mood and BD severity.

Methods. Fifty-two participants were recruited from a geographically well-defined mental health service in Northern Ireland. The AM test, self-report measures of lifetime experience of trauma, childhood trauma, and depression were administered. Severity of BD was estimated utilizing a systematic tool for reviewing all available clinical data of participants.

Results. A high prevalence of trauma was found. A total of 94.2% (49/52) of participants reported experiencing a traumatic event in either childhood or adulthood. AM specificity was significantly lower than previous reports of such in major depression. However, whilst childhood trauma predicted current inter-episode depressive mood, childhood trauma was not predictive of BD severity or AM specificity. Moreover, the association between childhood trauma and depressed mood was not moderated by AM specificity.

Conclusions. The findings of this study suggest a relationship between early psychosocial adversity and current inter-episode depressive mood in BD. In addition, levels of overgeneral AM are similar to that reported for depression, but are unrelated to childhood trauma, current inter-episode depressive mood, or BD severity. Clinical implications include the importance of routine assessment of trauma in BD and the need for adjunctive evidenced-based psychological therapies.

Endocrine disrupting effects of ochratoxin A at the level of nuclear receptor activation and steroidogenesis

Ochratoxin A (OTA) is a mycotoxin and extrolite of fungi which has been reported in a range of foods. This study uses mammalian reporter gene assays (RGAs) with natural steroid receptors and the H295R steroidogenesis assay to assess the endocrine disrupting activity of OTA.

At the receptor level, OTA (within a concentration range of 0.25–2500 ng/ml) did not induce an agonistic response in an oestrogen, androgen, progestagen or glucocorticoid RGA. An antagonistic effect was observed in all of the RGAs at the highest concentration tested (2500 ng/ml). However, while there was no significant cytotoxic effect observed in the MTT (thiazolyl blue tetrazolium bromide) cell viability assay at this concentration, there was a corresponding change in cell morphology which may be related to the resulting antagonistic effect.

At the hormone production level, H295R cells were used as a steroidogenesis model and exposed to OTA (within a concentration range of 0.1–1000 ng/ml). Treatment of the cells with 1000 ng/ml OTA increased the production of estradiol (117 ± 14 ng/ml) over 3 times that of the solvent control (36 ± 9 pg/ml). Western blotting confirmed an increase in aromatase protein.

Overall the results indicate that OTA does not appear to interact with steroid receptors but has the potential to cause endocrine disruption by interfering with steroidogenesis. This is the first study identifying the effect OTA may have on production of the steroid hormone estradiol.