THE CORRECTED PRIMARY STRUCTURE OF CHICKEN (AVIAN) PANCREATIC-POLYPEPTIDE

Chicken (avian) pancreatic polypeptide was the first member of the pancreatic polypeptide (PP)/neuropeptide Y (NPY) superfamily to be discovered and structurally-characterised. In this 36 amino acid residue, C-terminally amidated peptide, residues 22 and 23 were identified as Asp and Asn, respectively. However, sequencing of chicken PP using modem automated gas-phase sequencing technology has revealed that the original primary structure is incorrect in that residue 22 is Asn and that residue 23 is Asp. After digestion of chicken PP with endoproteinase Asp-N, fragments of chicken PP corresponding in molecular mass to residues 16-22 and 23-36, were unequivocally identified. The corrected primary structure of chicken PP is therefore: Gly-Pro-Ser-Gln-Pro-Thr-Tyr-Pro-Gly-Asp-Asp-Ala-Pro-Val-Glu-Asp-Leu-Ile-Arg-Phe-Tyr-Asn-Asp-Leu-Gln-Gln-Tyr-Leu-Asn-Val-Val-Thr-Arg-His-Arg-Tyr-NH2.

RABBIT PANCREATIC-POLYPEPTIDE

1. In almost all studies involving localization or quantitation of regulatory peptides, an essential prerequisite is the generation of specific antisera in rabbits. Despite this almost universal practice, the primary structures of some established regulatory peptides, such as pancreatic polypeptide (PP), of the rabbit, remain unknown.

A CYTOCHEMICAL STUDY OF THE NERVOUS-SYSTEM OF THE PROTEOCEPHALIDEAN CESTODE, PROTEOCEPHALUS-POLLANICOLA

The localization and distribution of cholinergic, serotoninergic and peptidergic nerve elements in the proteocephalidean tapeworm, Proteocephalus pollanicola, have been investigated by enzyme histochemistry, and by an indirect immunofluorescence technique interfaced with confocal scanning laser microscopy. Cholinesterase (ChE) activity was localized in the major components of the central nervous system (CNS) and the peripheral nervous system (PNS), including the innervation of the reproductive structures of the worm. Serotoninergic (5-HT) nerves were found in the paired cerebral ganglia, transverse commissure and in the 10 longitudinal nerve cords. Antisera to 17 mammalian regulatory peptides and the invertebrate peptide FMRFamide have been used to explore the peptidergic nervous system of the worm. The most extensive immunostaining occurred with antisera raised to members of the neuropeptide Y superfamily, namely neuropeptide Y (NPY), peptide YY (PYY) and pancreatic polypeptide (PP). In all cases, intense immunoreactivity was found in numerous cell bodies and fibres of both the CNS and PNS, including the innervation of the reproductive apparatus. FMRFamide antisera stained the same structures to a comparable degree as those raised to the NPY superfamily. Cholinergic and peptidergic elements were much more prevalent within the CNS, while the serotoninergic nerve fibres tended to dominate in the PNS. The overlap obtained in staining patterns for the peptidergic and cholinergic components suggests that there may be a certain amount of co-localization of peptides with small-molecule transmitter substances in the same neurone. Weak staining for the tachykinin, substance P and for calcitonin gene-related peptide(CGRP) was confined to the major longitudinal nerve cords.

THE PRIMARY STRUCTURE OF PANCREATIC-POLYPEPTIDE FROM A PRIMITIVE INSECTIVOROUS MAMMAL, THE EUROPEAN HEDGEHOG (ERINACEOUS-EUROPAEUS)

Pancreatic polypeptide (PP) has been isolated from extracts of the pancreas of the European hedgehog (Erinaceous europaeus) which is a representative of the order Insectivora, deemed to be the most primitive group of placental mammals. Pancreatic tissues were extracted in acidified ethanol and the peptide was purified chromatographically using a PP C-terminal hexapeptide amide specific radioimmunoassay to monitor purification. Two major PP-immunoreactive peptides were baseline-resolved following the final analytical reverse phase HPLC fractionation. Each was separately subjected to plasma desorption mass spectroscopy (PDMS) and gas-phase sequencing. The molecular masses of each peptide were similar: (I) 4237.6 +/- 4 Da and (II) 4238.2 +/- 4 Da. The full primary structures of each peptide were deduced and these were identical: VPLEPVYPGDNATPEQMAHYAAELRRYINMLTRPRY. The peptides were deemed to be amidated due to their full molar cross-reactivity with the amide-requiring PP antiserum employed in radioimmunoassay. The molecular mass (4233.8 Da) calculated from the sequence was in close agreemeent with PDMS estimates and the reason for the different retention times of each peptide is unknown at present. Hedgehog PP exhibits only 2 unique amino acid substitutions, at positions 1 (Val) and 19 (His), when compared with other mammalian analogues.

Genome-wide profiling of methylation identifies novel targets with aberrant hyper-methylation and reduced expression in low-risk myelodysplastic syndromes

Gene expression profiling signatures may be used to classify the subtypes of Myelodysplastic syndrome (MDS) patients. However, there are few reports on the global methylation status in MDS. The integration of genome-wide epigenetic regulatory marks with gene expression levels would provide additional information regarding the biological differences between MDS and healthy controls. Gene expression and methylation status were measured using high-density microarrays. A total of 552 differentially methylated CpG loci were identified as being present in low-risk MDS; hypermethylated genes were more frequent than hypomethylated genes. In addition, mRNA expression profiling identified 1005 genes that significantly differed between low-risk MDS and the control group. Integrative analysis of the epigenetic and expression profiles revealed that 66.7% of the hypermethylated genes were underexpressed in low-risk MDS cases. Gene network analysis revealed molecular mechanisms associated with the low-risk MDS group, including altered apoptosis pathways. The two key apoptotic genes BCL2 and ETS1 were identified as silenced genes. In addition, the immune response and micro RNA biogenesis were affected by the hypermethylation and underexpression of IL27RA and DICER1. Our integrative analysis revealed that aberrant epigenetic regulation is a hallmark of low-risk MDS patients and could have a central role in these diseases.

Architecture and Construction Management at Queens University Belfast:Regional Cultural Centre

The Regional Cultural Centre in Letterkenny is a new 2000sqm arts center containing theatre, galleries, workshops and ancillary offices. The site is set back from the street, on high ground with good views. The form and envelope of the building was derived from geometrically connecting the site with the town’s two other main public buildings, the Cathedral (1901) and new Civic Offices (2002, also designed by MacGabhann Architects). This geometrical connection or vectors informed the geometry and shape of the building. This urban matrix of geometrically connecting three corner stones of society, namely the ecclesiastical headquarters, the administrative head quarters and the art centre helps to improve the town planning and urban design of the disparate and chaotic development that Letterkenny has become.
The large cantilever, which houses a 300sqm gallery, is aligned towards the Civic Offices, marks the entrance, and signifies a change of direction of the pedestrian route past the building, like a modern day obelisk.
The circulation routes and stairs internally provide views towards the civic offices and cathedral, thus reinforcing the connection between the three buildings and helps visitors make some sense of Letterkenny as an urban center. The main stairs and vertical circulation are contained behind the large glazed foyer, which is framed to be viewed externally like a proscenium stage, with visitors to the building passively acting their routes through the building.

Efficacy, safety and effect on biomarkers of AZD9668 in cystic fibrosis

The aim of this study was to evaluate the safety and effect on clinical outcomes and biomarkers of inflammation and tissue damage of the neutrophil elastase inhibitor AZD9668 (60 mg twice daily orally for 4 weeks) in cystic fibrosis. This was a randomised, double-blind, placebo-controlled study. Primary outcome measures were sputum neutrophil count, lung function, 24-h sputum weight, BronkoTest® diary card data and health-related quality-of-life (revised cystic fibrosis quality-of-life questionnaire). Secondary end-points included sputum neutrophil elastase activity, inflammatory biomarkers in sputum and blood, urine and plasma desmosine (an elastin degradation marker), AZD9668 levels and safety parameters (adverse events, routine haematology, biochemistry, electrocardiogram and sputum bacteriology). 56 patients were randomised, of which 27 received AZD9668. There was no effect for AZD9668 on sputum neutrophil counts, neutrophil elastase activity, lung function or clinical outcomes, including quality of life. In the AZD9668 group, there was a trend towards reduction in sputum inflammatory biomarkers with statistically significant changes in interleukin-6, RANTES and urinary desmosine. The pattern of adverse events was similar between groups. Consistent reductions in sputum inflammatory biomarkers were seen in the AZD9668 group, and reduction in urinary desmosine suggests that AZD9668 impacts elastin cleavage by neutrophil elastase.

flp-32 Ligand/Receptor Silencing Phenocopy Faster Plant Pathogenic Nematodes

Restrictions on nematicide usage underscore the need for novel control strategies for plant pathogenic nematodes such as Globodera pallida (potato cyst nematode) that impose a significant economic burden on plant cultivation activities. The nematode neuropeptide signalling system is an attractive resource for novel control targets as it plays a critical role in sensory and motor functions. The FMRFamide-like peptides (FLPs) form the largest and most diverse family of neuropeptides in invertebrates, and are structurally conserved across nematode species, highlighting the utility of the FLPergic system as a broad-spectrum control target. flp-32 is expressed widely across nematode species. This study investigates the role of flp-32 in G. pallida and shows that: (i) Gp-flp-32 encodes the peptide AMRNALVRFamide; (ii) Gp-flp-32 is expressed in the brain and ventral nerve cord of G. pallida; (iii) migration rate increases in Gp-flp-32-silenced worms; (iv) the ability of G. pallida to infect potato plant root systems is enhanced in Gp-flp-32-silenced worms; (v) a novel putative Gp-flp-32 receptor (Gp-flp-32R) is expressed in G. pallida; and, (vi) Gp-flp-32R silenced worms also display an increase in migration rate. This work demonstrates that Gp30 flp-32 plays an intrinsic role in the modulation of locomotory behaviour in G. pallida, and putatively interacts with at least one novel G-protein coupled receptor (Gp-flp-32R). This is the first functional characterisation of a parasitic nematode FLP-GPCR. © 2013 Atkinson et al.

Towards a virtual testing environment for composite aerostructures

The emergence of an all-composite passenger airframe marks a major advance in the development of aerostructures. Underpinning this milestone is over four decades of intensive research in this area. Nonetheless, the first-generation of composite aerostructures is very conservative. This paper will discuss the need for the development of a virtual testing capability to enable better exploitation of the material's full potential in future designs. Recent progress, by the author, in this area is presented followed by a discussion of current limitations and opportunities for further research.

Glacial to paraglacial history and forest recovery in the Oglio glacier system (Italian Alps) between 26 and 15 ka cal BP

The integrated stratigraphic, radiocarbon and palynological record from an end-moraine system of the Oglio valley glacier (Italian Alps), propagating a lobe upstream in a lateral reach, provided evidence for a complete cycle of glacial advance, culmination and withdrawal during the Last Glacial Maximum and early Lateglacial. The glacier culminated in the end moraine shortly after 25.8 +/- 0.8 ka cal BP, and cleared the valley floor 18.3-17.2 +/- 0.3 ka cal BP. A primary paraglacial phase is then recorded by fast progradation of the valley floor.

As early as 16.7 +/- 0.3 ka cal BP, early stabilization of alluvial fans and lake filling promoted expansion of cembran pine. This is an unprecedented evidence of direct tree response to depletion of paraglacial activity during the early Lateglacial, and also documents the cembran pine survival in the mountain belt of the Italian Alps during the last glaciation. Between 16.1 and 14.6 +/- 0.5 ka cal BP, debris cones emplacement points to a moisture increase favouring tree Betula and Pinus sylvestris-mugo. A climate perturbation renewed paraglacial activity. According to cosmogenic ages on glacial deposits and AMS radiocarbon ages from lake records in South-Eastern Alps such phase compares favourably with the Gschnitz stadial and with the oscillations recorded at lakes Ragogna. Langsee and Jeserzersee, most probably forced by the latest freshening phases of the Heinrich Event 1.

A further sharp pine rise marks the subsequent onset of Bolling interstadial. The chronology of the Oglio glacier compares closely with major piedmont glaciers on the Central and Eastern Alpine forelands. On the other hand, the results of the present study imply a chronostratigraphic re-assessment of the recent geological mapping of the Central Italian Alps. (C) 2012 Elsevier Ltd. All rights reserved.

How standardised patients award global scores in OSCEs: a qualitative study

Standardized patients (SPs) are often asked to award global scores on the humanistic aspects of a candidate’s performance in an OSCE. However, little is known about the process by which SPs arrive at their mark.
Five focus groups of SPs, using a convenience sample, were used to collect data until saturation. Thematic analysis was carried out independently by three researchers using a grounded theory approach.

Four major themes contributed to their decision-making process: environment, relationships within the exam, preparedness for the task and expectations of the student’s performance. Environmental factors included the station itself, the rating scale and examiner fatigue. Relationship factors included first impressions, the sense of purpose derived from examining and a tendency to mirror the examiner’s reaction. Factors relating to preparedness for task involved experience as an SP and technical aspects, such as the need for calibration. Lastly, expectations of performance were related to preconceptions about what makes a ‘good’ student, including their level of studies, appearance and technical performance.

In assessing students, SPs drew on their wider attitudes and experiences. SPs did not limit their assessment to humanistic traits but often included technical performance. Thus, SPs to some extent assessed a similar construct to examiners and this may help to explain the increased reliability associated with using SP scores. SP global scores are a useful adjunct but the process by which SPs award marks is complex and provides a challenge for training and standardisation.

Cooperativity of imprinted genes inactivated by acquired chromosome 20q deletions

Large regions of recurrent genomic loss are common in cancers; however, with a few well-characterized exceptions, how they contribute to tumor pathogenesis remains largely obscure. Here we identified primate-restricted imprinting of a gene cluster on chromosome 20 in the region commonly deleted in chronic myeloid malignancies. We showed that a single heterozygous 20q deletion consistently resulted in the complete loss of expression of the imprinted genes L3MBTL1 and SGK2, indicative of a pathogenetic role for loss of the active paternally inherited locus. Concomitant loss of both L3MBTL1 and SGK2 dysregulated erythropoiesis and megakaryopoiesis, 2 lineages commonly affected in chronic myeloid malignancies, with distinct consequences in each lineage. We demonstrated that L3MBTL1 and SGK2 collaborated in the transcriptional regulation of MYC by influencing different aspects of chromatin structure. L3MBTL1 is known to regulate nucleosomal compaction, and we here showed that SGK2 inactivated BRG1, a key ATP-dependent helicase within the SWI/SNF complex that regulates nucleosomal positioning. These results demonstrate a link between an imprinted gene cluster and malignancy, reveal a new pathogenetic mechanism associated with acquired regions of genomic loss, and underline the complex molecular and cellular consequences of "simple" cancer-associated chromosome deletions.

Molecular diagnosis of the myeloproliferative neoplasms: UK guidelines for the detection of JAK2 V617F and other relevant mutations

Molecular genetic assays for the detection of the JAK2 V617F (c.1849G>T) and other pathogenetic mutations within JAK2 exon 12 and MPL exon 10 are part of the routine diagnostic workup for patients presenting with erythrocytosis, thrombocytosis or otherwise suspected to have a myeloproliferative neoplasm. A wide choice of techniques are available for the detection of these mutations, leading to potential difficulties for clinical laboratories in deciding upon the most appropriate assay, which can lead to problems with inter-laboratory standardization. Here, we discuss the most important issues for a clinical diagnostic laboratory in choosing a technique, particularly for detection of the JAK2 V617F mutation at diagnosis. The JAK2 V617F detection assay should be both specific and sensitive enough to detect a mutant allele burden as low as 13%. Indeed, the use of sensitive assays increases the detection rate of the JAK2 V617F mutation within myeloproliferative neoplasms. Given their diagnostic relevance, it is also beneficial and relatively straightforward to screen JAK2 V617F negative patients for JAK2 exon 12 mutations (in the case of erythrocytosis) or MPL exon 10 mutations (thrombocytosis or myelofibrosis) using appropriate assays. Molecular results should be considered in the context of clinical findings and other haematological or laboratory results.

Thinking Sexual Difference Through the Law of Rape

2013 marks 10 years since the Sexual Offences Act 2003 was passed. That Act made significant changes to the law of rape which appear now to have made very little difference to reporting, prosecution or conviction rates. This article argues that the Act has failed against its own measures because it remains enmeshed within a conceptual framework of sexual indifference in which woman continues to be constructed as man’s (defective) other. This construction both constricts the frame in which women’s sexuality can be thought and distorts the harm of rape for women. It also continues woman’s historic alienation from her own nature and denies her entitlement to a becoming in line with her own sexuate identity. Using Luce Irigaray’s critical and constructive frameworks, the article seeks to imagine how law might ‘cognize’ sexual difference and thus take the preliminary steps to a juridical environment in which women can more adequately understand and articulate the harm of rape.