Isolation, pharmacology and gene organization of KPSFVRFamide: A neuropeptide from Caenorhabditis elegans

To date, 53 peptides with C-terminal RFamides have been identified by the genome sequencing project in the nematode, Caenorhabditis elegans. In this study the FMRFamide-related peptide (FaRP) KPSFVRFamide (879.90 Da [MH](+)) was structurally characterized from extracts of the nematode, Caenorhabditis elegans. Two copies of KPSFVRFamide are encoded by a gene designated flp-9. RT-PCR identified a single cDNA product which was confirmed as flp-9 by sequence determination. Flp-9 cDNA was isolated from larval stages of C. elegans but was not detected-in adult worms, indicating that its expression is may be developmentally regulated. KPSFVRFamide displays sequence homology to the nematode peptide, KPNFIRFamide (PF4). The physiological effects of KPSFVRFamide, PF4 and the chimeras, KPNFVRFamide and KPSFIRFamide, were measured on body wall muscle and the vagina vera of the parasitic nematode, Ascaris suum. KPNFVRFamide and KPNFIRFamide had Cl--dependent inhibitory activity on innervated and denervated muscle-preparations, whereas KPSFVRFamide and KPSFIRFamide did not elicit a detectable physiological effect. Although all 4 peptides had inhibitory effects on the vagina vera, KPSFVRFamide and KPSFIRFamide (threshold, greater than or equal to 0.1 mu M) were less potent than KPNFVRFamide and KPNFIRFamide (threshold, greater than or equal to 10 nM). (C) 1999 Academic Press.

THE ORGANIZATION OF THE NERVOUS-SYSTEM IN PLATHELMINTHES - THE NEUROPEPTIDE F-IMMUNOREACTIVE PATTERN IN CATENULIDA, MACROSTOMIDA, PROSERIATA

The organization of the nervous system of Archilopsis unipunctata Promonotus schultzei and Paramonotus hamatus (Monocelididae, Proseriata) and Stenostomum leucops (Catenulida) and Microstomum lineare (Macrostomida) was studied by immunocytochemistry, using antibodies to the authentic flatworm neuropeptide F (NPF) (Moniezia expansa). The organization of the nervous system of the Monocelididae was compared to that of the nervous system of Bothriomolus balticus (Otoplanidae), a previously studied species of another family of the Proseriata. The results show that the main nerve cords (MCs), independent of lateral or ventral position in the Monocelididae and the Otoplanidae, correspond to each other. The study also confirms the status of the lateral cords as main cords (MCs) in S. leucops and M. lineare. Common for MCs in the members of the investigated taxa are the following features: MCs consist of many fibres, originate from the brain and are adjoined to 5-HT-positive neurons. In Monocelididae and Otoplanidae, the MCs additionally have the same type of contact to the pharyngeal nervous system. Also common for both proseriate families is the organization of the two lateral nerve cords, with weaker connections to the brain, and the pair of dorsal cords running above the brain. The organization of the minor cords differs. The Monocelididae have a pair of thin ventral cords forming a mirror image of the dorsal pair. Furthermore, an unpaired ventral medial cord connecting medial commissural cells was observed in P. schultzei. Marginal nerve cords, observed in Otoplanidae, are absent in Monocelididae. All minor nerve cords are closely connected to the peripheral nerve plexus. The postulated trends of condensation of plexal fibres to cords and/or the flexibility of the peripheral nerve plexus are discussed. In addition, the immunoreactivity (IR) pattern of NPF was compared to the IR patterns of the neuropeptide RFamide and the indoleamine, 5-HT (serotonin). Significant differences between the distribution of IR to NPF and to 5-HT occur. 5-HT-IR dominates in the submuscular and subepidermal plexuses. In the stomatogastric plexus of M. lineare, only peptidergic IR is observed in the intestinal nerve net. The distribution of NPF-IR in fibres and cells of the intestinal wall in M. lineare indicates a regulatory function for this peptide in the gut, while a relationship with ciliary and muscular locomotion is suggested for the 5-HT-IR occurring in the subepidermal and submuscular nerve plexuses. In M. lineare, the study revealed an NPF- and RFamide-positive cell pair, marking the finished development of new zooids. This finding indicates that constancy of these cells is maintained in this asexually reproducing and regenerating species.

Organization of mammary epithelial cells into 3D acinar structures requires glucocorticoid and JNK signaling

Mammary epithelial cells cultured on a concentrated laminin-rich extracellular matrix formed 3D acinar structures that matured to polarized monolayers surrounding a lumen. In the absence of glucocorticoids mature acinus formation failed and the expression of an acinus-associated, activator protein 1 (AP1) and nuclear factor kappaB transcription factor DNA-binding profile was lost. Treatment with the JNK inhibitor, SP600125, caused similar effects, whereas normal organization of the mammary epithelial cells as acini caused JNK activation in a glucocorticoid-dependent manner. The forming acini expressed BRCA1, GADD45beta, MEKK4, and the JNK activating complex GADD 45beta-MEKK4 in a glucocorticoid-dependent fashion. JNK catalyzed phosphorylation of c-Jun was also detected in the acini. In addition, expression of beta4 integrin and in situ occupation of its promoter by AP1 components, c-Jun and Fos, was glucocorticoid dependent. These results suggest that glucocortocoid signaling regulates acinar integrity through a pathway involving JNK regulation of AP1 transcription factors and beta4 integrin expression.

Measuring party organization: The vertical dimension of the multilevel organization of statewide parties in Spain and the UK

Over the past few years, attention to the role of state-wide political parties in multi-level polities has increased in recognition of their linkage function between levels of government, as these parties compete in both state-wide and regional elections across their countries. This article presents a coding scheme designed to describe the relationship between central and regional levels of state-wide parties. It evaluates the involvement of the regional branches in central decision-making and their degree of autonomy in the management of regional party affairs. This coding scheme is applied to state-wide parties in Spain (the socialist PSOE and the conservative Partido Popular) and in the UK (Labour, the Conservatives and the Liberal Democrats). It is an additional tool with which to analyse party organization and it facilitates the comparison of parties across regions and in different countries.

Re-organisation of the cytoskeleton during developmental programmed cell death in Picea abies embryos

Cell and tissue patterning in plant embryo development is well documented. Moreover, it has recently been shown that successful embryogenesis is reliant on programmed cell death (PCD). The cytoskeleton governs cell morphogenesis. However, surprisingly little is known about the role of the cytoskeleton in plant embryogenesis and associated PCD. We have used the gymnosperm, Picea abies , somatic embryogenesis model system to address this question. Formation of the apical-basal embryonic pattern in P. abies proceeds through the establishment of three major cell types: the meristematic cells of the embryonal mass on one pole and the terminally differentiated suspensor cells on the other, separated by the embryonal tube cells. The organisation of microtubules and F-actin changes successively from the embryonal mass towards the distal end of the embryo suspensor. The microtubule arrays appear normal in the embryonal mass cells, but the microtubule network is partially disorganised in the embryonal tube cells and the microtubules disrupted in the suspensor cells. In the same embryos, the microtubule-associated protein, MAP-65, is bound only to organised microtubules. In contrast, in a developmentally arrested cell line, which is incapable of normal embryonic pattern formation, MAP-65 does not bind the cortical microtubules and we suggest that this is a criterion for proembryogenic masses (PEMs) to passage into early embryogeny. In embryos, the organisation of F-actin gradually changes from a fine network in the embryonal mass cells to thick cables in the suspensor cells in which the microtubule network is completely degraded. F-actin de-polymerisation drugs abolish normal embryonic pattern formation and associated PCD in the suspensor, strongly suggesting that the actin network is vital in this PCD pathway.

Re-analysis of microarray data reveals insights into altered transcriptional activity of TH17 and TReg signalling in psoriasis

Identifying differential expression of genes in psoriatic and healthy skin by microarray data analysis is a key approach to understand the pathogenesis of psoriasis. Analysis of more than one dataset to identify genes commonly upregulated reduces the likelihood of false positives and narrows down the possible signature genes. Genes controlling the critical balance between T helper 17 and regulatory T cells are of special interest in psoriasis. Our objectives were to identify genes that are consistently upregulated in lesional skin from three published microarray datasets. We carried out a reanalysis of gene expression data extracted from three experiments on samples from psoriatic and nonlesional skin using the same stringency threshold and software and further compared the expression levels of 92 genes related to the T helper 17 and regulatory T cell signaling pathways. We found 73 probe sets representing 57 genes commonly upregulated in lesional skin from all datasets. These included 26 probe sets representing 20 genes that have no previous link to the etiopathogenesis of psoriasis. These genes may represent novel therapeutic targets and surely need more rigorous experimental testing to be validated. Our analysis also identified 12 of 92 genes known to be related to the T helper 17 and regulatory T cell signaling pathways, and these were found to be differentially expressed in the lesional skin samples.

Ages of 24 widespread tephras erupted since 30,000 years ago in New Zealand, with re-evaluation of the timing and palaeoclimatic implications of the Lateglacial cool episode recorded at Kaipo bog

Tephras are important for the NZ-INTIMATE project because they link all three records comprising the composite inter-regional stratotype developed for the New Zealand climate event stratigraphy (NZ-CES). Here we firstly report new calendar ages for 24 widespread marker tephras erupted since 30,000 calendar (cal.) years ago in New Zealand to help facilitate their use as chronostratigraphic dating tools for the NZ-CES and for other palaeoenvironmental and geological applications. The selected tephras comprise 12 rhyolitic tephras from Taupo, nine rhyolitic tephras from Okataina, one peralkaline rhyolitic tephra from Tuhua, and one andesitic tephra each from Tongariro and Egmont/Taranaki volcanic centres. Age models for the tephras were obtained using three methods: (i) C-based wiggle-match dating of wood from trees killed by volcanic eruptions (these dates published previously); (ii) flexible depositional modelling of a high-resolution C-dated age-depth sequence at Kaipo bog using two Bayesian-based modelling programs, Bacon and OxCal's P_Sequence function, and the IntCal09 data set (with SH offset correction-44±17yr); and (iii) calibration of C ages using OxCal's Tau_Boundary function and the SHCal04 and IntCal09 data sets. Our preferred dates or calibrated ages for the 24 tephras are as follows (youngest to oldest, all mid-point or mean ages of 95% probability ranges): Kaharoa AD 1314±12; Taupo (Unit Y) AD 232±10; Mapara (Unit X) 2059±118cal.yrBP; Whakaipo (Unit V) 2800±60cal.yrBP; Waimihia (Unit S) 3401±108cal.yrBP; Stent (Unit Q) 4322±112cal.yrBP; Unit K 5111±210cal.yrBP; Whakatane 5526±145cal.yrBP; Tuhua 6577±547cal.yrBP; Mamaku 7940±257cal.yrBP; Rotoma 9423±120cal.yrBP; Opepe (Unit E) 9991±160cal.yrBP; Poronui (Unit C) 11,170±115cal.yrBP; Karapiti (Unit B) 11,460±172cal.yrBP; Okupata 11,767±192cal.yrBP; Konini (bed b) 11,880±183cal.yrBP; Waiohau 14,009±155cal.yrBP; Rotorua 15,635±412cal.yrBP; Rerewhakaaitu 17,496±462cal.yrBP; Okareka 21,858±290cal.yrBP; Te Rere 25,171±964cal.yrBP; Kawakawa/Oruanui 25,358±162cal.yrBP; Poihipi 28,446±670cal.yrBP; and Okaia 28,621±1428cal.yrBP.Secondly, we have re-dated the start and end of the Lateglacial cool episode (climate event NZce-3 in theNZ-CES), previously referred to as the Lateglacial climate reversal, as defined at Kaipo bog in eastern North Island, New Zealand, using both Bacon and OxCal P_Sequence modelling with the IntCal09 data set. The ca1200-yr-long cool episode, indicated by a lithostratigraphic change in the Kaipo peat sequence to grey mudwith lowered carbon content, and a high-resolution pollen-derived cooling signal, began 13,739±125cal.yrBP and ended 12,550±140cal.yrBP (mid-point ages of the 95% highest posterior density regions, Bacon modelling). The OxCal modelling, generating almost identical ages, confirmed these ages. The Lateglacial cool episode (ca 13.8-12.6cal.kaBP) thus overlaps a large part of the entire Antarctic Cold Reversal chronozone (ca 14.1-12.4cal.kaBP or ca 14.6-12.8cal.kaBP), and an early part of the Greenland Stadial-1 (Younger Dryas) chronozone (ca 12.9-11.7cal.kaBP). The timing of the Lateglacial cool episode at Kaipo is broadly consistent with the latitudinal patterns in the Antarctic Cold Reversal signal suggested for the New Zealand archipelago from marine and terrestrial records, and with records from southern South America. © 2012 Elsevier Ltd.

Risk and Reward: Safer systems of work practice.

To develop and implement a risk assessment process for all unlicenced medicinal products in use within the Belfast City Hospital.
Over half 65% of the unlicenced medicinal products currently in use were rated low or minor risk and therefore required no recording upon supply.
This has greatly improved the way unlicensed medicines are prescribed, procured, supplied and administered within the Belfast City Hospital.