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In strong collaboration with the oil company consortium, contractor and their subcontractors successfully designed and installed a novel Water Intake Riser (WIR) on Prelude FLNG, the world's largest FLNG facility. The scope involved transforming a concept into reality by close collaboration between the contractor and client team at various offices and worksites from project start to offshore installation. Preparation for this operation included activities at various locations around the world such as Factory Acceptance Testing in Norway, USA, UK and France. The final Site Integration Testing was performed at the FLNG in South Korea before Prelude FLNG left the yard.Designing a Water Intake Riser (WIR) at this scale capable of surviving 10,000 years cyclone with a design life of 25 years has raised a variety of engineering challenges including; riser head-loss, loadings under survival events, novel flying leads and umbilicals, fatigue of key element such as chains, spacer-frames and riser girth welds. Full-scale tests were performed on critical components of the WIR for example the rubber hose, chain, and connectors. In addition, the system was tested at scale in model scale basins.Offshore execution in Australia involved the installation of large diameter steel pipe sections assembled with merlin connectors and the largest submersible pumps of their kind inside the FLNG Hull. Bespoke installation aids were used such as an over-the-side Riser Assembly Tower. Successful execution required maintaining Prelude FLNG at a favorable heading to minimize motions and achieve millimeter precision pipe assembly.

The Prelude FLNG facility anchoring system is based on a turret with 16 anchor legs, each connected to driven anchor piles in generally very weak and variable carbonate soils. Driven piles have a chequered history offshore Australia, due to the very low skin friction that is typically mobilized. At Prelude, the combination of weak soils with some harder layers and low expected skin friction led to numerous installation challenges such as free-fall, overpenetration, soil blow-out and plug depression risks. Mitigating these risks was achieved in a joint risk-based approach resulting in the unique combination of floating plate parachute, bearing V-shaped cruciform and "pushing" weight tool and installation methodology. Post-treatment of the lifting crane data allowed taking benefit of the experienced soil behavior during the campaign to improve the methods with every pile, leading to the successful installation of the 16 anchors. It also improved the knowledge of the heterogeneous Australian carbonate soils including sand/silt/clay layers through comparison with geotechnical original geotechnical investigations and laboratory tests. This paper describes the various design and installation challenges and the novel solutions which were applied to manage the substantial risk of failure to install these anchor piles. The successful installation and associated data acquired is unique and have led to invaluable improvements in the understanding of pile-soil interaction in carbonate soils.

In Central European temperate forests, the host tree identity is one factor that determines the diversity of saproxylic organisms. These forests have been affected by humans for millennia, in contrast to the Hyrcanian forests south of the Caspian Sea, with their numerous old-growth features and endemic species. How the tree host species in this temperate relict of Tertiary forests affects saproxylic biodiversity is unknown, as an inventory of the saproxylic fauna present is incomplete, and new species are still being described. To analyze the importance of four dominant tree species in this region (Fagus orientalis, Quercus castaneifolia, Alnus subcordata, and Carpinus betulus), we sampled saproxylic beetles and true bugs with flight-interception traps at five dead trees per host species. We identified 361 beetle and 7 true bug species; of these, 268 were saproxylic, 56 were endemic, 11 were Central European “primeval-forest relicts”, and at least 16 species were undescribed. Rarefaction-extrapolation curves indicated higher species richness of saproxylic beetles and true bugs on F. orientalis and C. betulus than on A. subcordata and Q. castaneifolia, but ordination showed that communities were most distinct on Q. castaneifolia. Higher species richness on C. betulus and F. orientalis compared to Q. castaneifolia was unexpected based on literature but is in line with recent findings of experimental studies from Central Europe. Our results underline previous findings on the importance of moribund trees and dead wood in the Hyrcanian forests, and thereby support current efforts of Iranian authorities to stop the removal of veteran trees. The complementary fauna on oak trees compared to the other three tree species illustrates the importance of fragments of the almost destroyed oak forests along the coastline for forest biodiversity conservation in the Hyrcanian region.

Disziplinübergreifendes Forschungsdatenmanagement für Hochschulbibliotheken und Projekte zu vereinfachen und zu etablieren – das ist das Ziel von RADAR. Im Sommer 2016 geht mit ‚RADAR – Research Data Repository‘ ein Service an den Start, der Forschenden, Institutionen verschiedener Fachdisziplinen und Verlagen eine generische Infrastruktur für die Archivierung und Publikation von Forschungsdaten anbietet. Zu den Dienstleistungen gehören u. a. die Langzeitverfügbarkeit der Daten mit Handle oder Digital Object Identifier (DOI), ein anpassbares Rollen- und Zugriffsrechtemanagement, eine optionale Peer-Review-Funktion und Zugriffsstatistiken. Das Geschäftsmodell ermutigt Forschende, die anfallenden Nutzungsgebühren des Repositoriums in Drittmittelanträge und Datenmanagementpläne zu integrieren. Publizierte Daten stehen als Open Data zur Nachnutzung wie etwa Data Mining, Metadaten-Harvesting und Verknüpfung mit Suchportalen zur Verfügung. Diese Vernetzung ermöglicht ein nachhaltiges Forschungsdatenmanagement und die Etablierung von Dateninfrastrukturen wie RADAR., The goal of RADAR is to simplify and establish inter-disciplinary research data management for university libraries and projects. In summer 2016‚ ‘RADAR – Research Data Repository‘ starts as a service that offers researchers, institutions of different disciplines and publishers a generic infrastructure for archiving and publishing their research data. Among others, services are long-term data availability with Handle or Digital Object Identifier (DOI), an adaptable role and access rights management, an optional peer review function and access statistics. The business model encourages researchers to integrate arising charges for using the repository into applications for Third-Party funding and data management plans. Published data are available as Open Data to be used by Data Mining, metadata harvesting or linking with search portals. This interlinking enables a sustainable research data management and the establishment of data infrastructures like RADAR.

We study changes in the dynamics of polymer chains confined to cylindrical nanochannels within aluminum oxide membranes. Specifically, a proton time-domain NMR technique is used to assess the effect of transient wall contacts on the time-averaged orientational order of poly(butadiene) segments in melts with different molecular weights (MW). Previous work has evidenced that the weakly interacting polymer, residing in ∼100 μm long, 20 and 60 nm wide channels, shows no significant confinement-related changes in the segmental (α) relaxation time and only weak (less than a factor of 2) changes in the micrometer-scale diffusivity. In the relevant temperature range above 340 K, we here use samples with pores oriented at different angles with respect to the main magnetic field to study the macroscopic anisotropy of segmental rotations and the effect of slower motions in regimes III and IV of the tube model up to the milliseconds time scale. We show that the pore walls exert a significant orientation effect on the...

Thermoresponsive polymer gels exhibit pronounced swelling and deswelling upon changes in temperature, accompanied by dynamic concentration fluctuations that have been interpreted as critical opalescence. These fluctuations span lengthscales similar to that of static structures in the gels, such as the gel polymer-network meshsize (1–10 nm) and static polymer-network crosslinking inhomogeneities (10–1000 nm). To systematically investigate this overlay, we use droplet-based microfluidics and fabricate submillimeter-sized gel particles with varying static heterogeneity, as revealed on a molecular scale by proton NMR. When these microgels are probed by small-angle neutron scattering, the detection of dynamic fluctuations during the volume phase transitions is strongly perturbed by the co-existing static inhomogeneity. Depending of the type of data analysis employed, the temperature-dependent evolution of the correlation length associated to the dynamic fluctuations does or does not agree with predictions by the critical scaling theory. Only the most homogeneous sample of this study, prepared by controlled polymer crosslinking in droplet microfluidics, shows a diverging correlation length in agreement to the critical scaling theory independent of the specific approach of data analysis. These findings suggest that care must be taken about polymer-network heterogeneity when gel volume phase transitions are evaluated as critical phenomena. © 2015 Wiley Periodicals, Inc. J. Polym. Sci., Part B: Polym. Phys. 2015, 53, 1112–1122

A novel class of bolapolyphile (BP) molecules are shown to integrate into phospholipid bilayers and self-assemble into unique sixfold symmetric domains of snowflake-like dendritic shapes. The BPs comprise three philicities: a lipophilic, rigid, π-π stacking core; two flexible lipophilic side chains; and two hydrophilic, hydrogen-bonding head groups. Confocal microscopy, differential scanning calorimetry, XRD, and solid-state NMR spectroscopy confirm BP-rich domains with transmembrane-oriented BPs and three to four lipid molecules per BP. Both species remain well organized even above the main 1,2-dipalmitoyl-sn-glycero-3-phosphocholine transition. The BP molecules only dissolve in the fluid membrane above 70 °C. Structural variations of the BP demonstrate that head-group hydrogen bonding is a prerequisite for domain formation. Independent of the head group, the BPs reduce membrane corrugation. In conclusion, the BPs form nanofilaments by π stacking of aromatic cores, which reduce membrane corrugation and possibly fuse into a hexagonal network in the dendritic domains.

Changes in large-scale polymer diffusivity along interfaces, arising from transient surface contacts at the nanometer scale, are not well understood. Using proton pulsed-gradient NMR, we here study the equilibrium micrometer-scale self-diffusion of poly(butadiene) chains along ∼100 μm long, 20 and 60 nm wide channels in alumina, which is a system without confinement-related changes in segmental relaxation time. Unlike previous reports on nonequilibrium start-up diffusion normal to an interface or into particulate nanocomposites, we find a reduction of the diffusivity that appears to depend only upon the pore diameter but not on the molecular weight in a range between 2 and 24 kg/mol. We rationalize this by a simple volume-average model for the monomeric friction coefficient, which suggests a 10-fold surface-enhanced friction on the scale of a single molecular layer. Further support is provided by applying our model to the analysis of published data on large-scale diffusion in thin films.

Introduction: Complex aberrant karyotypes (CK, ≥3 cytogenetic aberrations, CA) are associated with an unfavorable prognosis and an increased AML transformation rate in MDS. However, even MDS with CK (CK-MDS) are heterogeneous in terms of genetic profile and prognosis. Recently, we demonstrated that a high number of CA as well as mutations in TP53 (TP53mut) are associated with increased risk in CK-MDS (Haase et al, 2019). However, as there is a strong association between CK-MDS and TP53mut, it is still a matter of debate whether the karyotype and TP53mut are prognostically independent genetic markers. Furthermore, loss of heterozygosity (LOH) of 17p13 (TP53LOH), due to loss of genetic material or to copy number neutral LOH (CN-LOH), is also associated with a poor prognosis. We here aimed to characterize TP53mut andTP53LOH in CK-MDS and to elucidate the impact of cytogenetics, TP53mut and TP53LOH on the outcome of CK-MDS. Methods: We included 178 pts with MDS (N=138), CMML (N=5) and secondary AML after MDS (AML with myelodysplasia-related changes, N=35), all with CK. The median precentage of bone marrow (bm) blasts was 11% (range: 0-90%). The median age was 72 yrs (range: 30-95 yrs). The male:female ratio was 1.23:1. The number of CA was determined by banding analysis in all cases. The karyotype was confirmed by multicolor FISH in 134 cases. TP53LOH was verified by FISH analysis of the TP53 locus in 17p13 (146 analyses) and/or molecular karyotyping (MK, 41 analyses). In 144 cases further FISH probes in addition to TP53 were used. TP53mut was identified by NGS (54 cases) or Sanger sequencing (124 cases). Follow-up data for survival analyses were available for 127 pts with MDS and oligoblastic AML with less than 30% bm blasts. Results: The median number of CA was 7 (range: 3-46), 98/178 pts (55%) showed a TP53mut (median VAF: 34%, range: 8-93%) and 64/178 (36%) a TP53LOH (median FISH clone size: 65%, range: 6-99%), including 9 pts with a CN-LOH in 17p13. The CN-LOH was either identified by MK (5/41 pts (12%) where MK was available showed a CN-LOH, 4/5 with TP53mut) or by NGS (4/54 pts (7%) where NGS was available showed a VAF >70% and normal TP53-FISH). In total, a TP53mut and/or a TP53LOH was identified in 116/178 pts (65%). Overall survival (OS) did not significantly differ between CK-MDS with TP53mut only, TP53LOH only, and TP53mut+TP53LOH (Fig.1). Therefore, we merged TP53mut and TP53LOH to TP53altered in all further analyses. Regarding the cytogenetic characterization of pts with TP53altered, the number of CA was significantly higher in pts with TP53altered than in pts with normal TP53 (median 9 CA (range: 3-46) vs 5 CA (range: 3-24), P The number of CA as well as the TP53 status contributed significantly to OS (Fig.2). The presence of anemia (Hb Conclusions: The presence of ≥5 CA is associated with reduced OS in CK-MDS. A TP53mut as well as a TP53LOH both further segregate outcome. The impact of the clone size of TP53mut and TP53LOH on survival is currently being evaluated. Our data imply that the TP53 status (TP53mut and/or TP53LOH) and the complexity of the karyotype are independent prognostic markers. Based on the presence of anemia, the TP53 status (TP53mut and/or TP53LOH), and the number of CA, the individual risk of CK-MDS can be estimated more accurately. This will allow to better tailor treatment decisions for individual pts with CA. Funding (FS): 2017 SGR 288-GRC Disclosures Germing: Jazz Pharmaceuticals: Honoraria; Novartis: Honoraria, Research Funding; Amgen: Honoraria; Celgene: Honoraria, Research Funding. Kaivers:Jazz Pharmaceuticals: Other: Travel Support. Kröger:Celgene: Honoraria, Research Funding; DKMS: Research Funding; JAZZ: Honoraria; Medac: Honoraria; Neovii: Honoraria, Research Funding; Novartis: Honoraria, Research Funding; Riemser: Research Funding; Sanofi-Aventis: Research Funding. Hertenstein:RS Media: Research Funding. Döhner:Daiichi: Honoraria; Jazz: Honoraria; Novartis: Honoraria; Celgene: Honoraria; Janssen: Honoraria; CTI Biopharma: Consultancy, Honoraria. Bug:Hexal: Membership on an entity's Board of Directors or advisory committees; Celgene Neovii: Other: travel grant; Novartis: Membership on an entity's Board of Directors or advisory committees, Research Funding; Gilead Sciences: Membership on an entity's Board of Directors or advisory committees, Other: Travel grants; Sanofi: Other: travel grants; Celgene: Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: travel grants; Jazz Pharmaceuticals: Honoraria; Pfizer: Membership on an entity's Board of Directors or advisory committees. Nickelsen:Roche Pharma AG: Membership on an entity's Board of Directors or advisory committees, Other: Travel Grants; Celgene: Membership on an entity's Board of Directors or advisory committees, Other: Travel Grant; Janssen: Membership on an entity's Board of Directors or advisory committees. Platzbecker:Celgene: Consultancy, Honoraria; Abbvie: Consultancy, Honoraria; Novartis: Consultancy, Honoraria.

We present a study of the structure of Tetra-PEG model networks, using proton multiple-quantum NMR at low field in combination with computer simulations. Tetra-PEG is a novel high-performance hydrogel designed by combination of two symmetric tetra-arm macromonomers. In contrast to conventional hydrogels, which are highly heterogeneous due to fixed concentration fluctuations, Tetra-PEG exhibits a much less heterogeneous microstructure as indicated by previous light and small-angle neutron scattering studies. Here, the local-scale connectivity inhomogeneities, i.e., the sol and dangling polymer chains as well as the typical network connectivity defects resulting from the A–B reaction of four-arm macromonomers, are quantified experimentally for the first time, studying as-prepared Tetra-PEG hydrogels, cross-linked at different polymer concentrations and stoichiometries. To this end, we developed a novel approach for the analysis of double-quantum buildup curves consisting of well-distinguishable components w...

Site-specific genetic engineering is a valuable tool for pharmaceutical research and development of biomedical models. Despite engineered nucleases allow targeted gene edition in a rather simple fashion; few reports are available so far on specific gene knock-in (KI) combined with engineered nucleases in domestic species. Here, we evaluated the possibility of inducing specific KI of cDNAs coding for proteins of pharmaceutical interest under the control of milk native promoter sequences, taking advantage of the TALEN system, both in ovine somatic cells and in zygotes. We designed 2 TALENs, targeting exons 1 and 5 of ovine β-lactoglobulin gene (BLG), respectively, and a homologous recombination vector (pHR), carrying recombinant human factor IX (rhFIX) flanked by homology arms contiguous to the TALEN target sites. In an initial set of experiments, 5 × 105 to 1 × 106 ovine fibroblasts were transfected with 1 μg of each TALEN mRNA, with or without 50 ng μL−1 pHR. The feasibility of inducing knock-out (KO) was confirmed by Cel1 assay. The deletion of the genomic region between TALEN target sites and the occurrence of HR in cell lysates were assessed by PCR. Also, 576 individual colonies were picked up and analyzed by PCR. The deletion of the region between TALEN target sites was achieved with 7.8% efficiency (45/576). The incidence of HR in cells was 0.5% (3/576), as detected by PCR. In order to evaluate the system in zygotes, laparoscopic AI was performed on synchronized and superovulated ewes. Zygotes were recovered 16 h after AI and cytoplasmically injected with (1) 5 ng μL−1 TALEN mix (2.5 ng μL−1 oaBLG T1.1 + 2.5 ng μL−1 oaBLG T5.1) (5TM); (2) 5 ng μL−1 TALEN mix + 25 ng μL−1 pHR-hFIX plasmid (5TM+25pRH); or (3) 15 ng μL−1 TALEN mix (7.5 ng μL−1 oaBLG T1.1 + 7.5 ng μL−1 oaBLG T5.1) + 50 ng μL−1 pHR-hFIX (15TM+50pRH). A non-injected control (NIC) was also included. Embryo analysis was conducted on whole-genome amplified DNA from blastocysts, followed by PCR and sequencing. Non-parametric Fisher test was applied to detect significant differences among treatments. Although blastocyst rates for NIC and 5TM did not statistically differ, 5TM+25pRH and 15TM+50pRH groups resulted in lower blastocysts rates than the NIC [P

Abstract The Arctic Operations Handbook Joint Industry Project (JIP) was set up in 2012 with a focus on the operational activities for transport and installation of fixed, floating and subsea units, as well as for dredging, trenching, pipe laying and floating oil and gas production in Arctic and cold weather conditions. The prime purpose of the JIP was to identify gaps in the existing standards and guidelines. Specific recommendations were subsequently proposed which will hopefully contribute to the development of internationally accepted standards and guidelines. The project addressed safety and sustainability of offshore operations in the Arctic. This investigation into existing rules, regulations, standards and guidelines was intended to provide a common understanding for the offshore industries. Work groups were formed to execute work packages which were judged to be of prime importance. Taking into account numerous aspects of the impact of the Arctic on various operations it was considered that the overall risk levels for such operations can be reduced by adopting the outline recommendations including, for instance, those related to weather monitoring and forecasting, environmental impacts, logistics, equipment preparation, vessel operations, training and health and safety management.

The incorporation of polymers or smaller complex molecules into lipid membranes allows for property modifications or the introduction of new functional elements. The corresponding molecular-scale details, such as changes in dynamics or features of potential supramolecular structures, can be studied by a variety of solid-state NMR techniques. Here, we review various approaches to characterizing the structure and dynamics of the guest molecules as well as the lipid phase structure and dynamics by different high-resolution magic-angle spinning proton and 13C NMR experiments as well as static 31P NMR experiments. Special emphasis is placed upon the incorporation of novel synthetic polyphilic molecules such as shape-persistent T- and X-shaped molecules as well as di- and tri-block copolymers. Most of the systems studied feature dynamic heterogeneities, for instance those arising from the coexistence of different phases; possibilities for a quantitative assessment are of particular concern.

Introduction: Alterations of TP53 (cytogenetic 17p13.1 deletions and molecular TP53 mutations) were reported to be frequent in pts with myelodysplastic syndromes (MDS) and complex abnormalities (≥3 clonal cytogenetic aberrations, CA) that represent around 15% of all MDS cases. It was suggested that pts with MDS and complex abnormalities may be further prognostically subdivided by the molecular TP53 mutation status (Bejar et al, ASH, 2014, abstract #532). In this study we investigated the frequency of different types of TP53 alterations, their cytogenetic profile and their clinical impact in the adverse cytogenetic MDS subgroup of complex abnormalities. We performed comprehensive cytogenetic and molecular genetic analysis focusing as well on the extent of cytogenetic instability. Methods: We included 105 pts (57 m/48 f; median 71 yrs, range, 47-95 yrs) with MDS (n=86) and sAML after MDS (n=19) with complex abnormalities in our study. A total of 56/89 (62.9%) pts had received azacitidine. Survival was censored at allogeneic stem cell transplantation (26/83; 31.3%). Pts were characterized by chromosome banding analysis, interphasefluorescence in situ hybridization (FISH) with a panel including a 17p13/TP53-covering probe, multicolor FISH (mFISH), Sanger sequencing of TP53 and SNP-array analysis (SNP-A). The extent of genetic imbalances was objectified by counting the number of CA, the number of cytogenetic fusions as shown by mFISH and the size of total genomic aberrations (TGA) measured by SNP-A in megabases (Mb). Results: A molecular TP53 mutation was found in 46/105 (43.8%) pts; a cytogenetic TP53 deletion in 38/105 (36.2%) pts. TP53 was not affected by a molecular mutation or a cytogenetic deletion in 44/105 (42.2%) pts, 23/105 (21.9%) pts were affected by combined TP53 alterations (molecular mutation and cytogenetic deletion), 23/105 (21.9%) pts by a molecular mutation only and 15/105 (14.3%) pts by a cytogenetic deletion only. The median number of CA was 6 (range, 3-41) in the entire cohort. Median overall survival for the entire cohort was 17 months. The degree of genomic imbalances was higher in pts with any TP53 alteration (molecular mutation and/or cytogenetic deletion) as compared to those without: The median number of CA was 8 (range, 3-41) vs. 4 (3-20) (P By univariate analysis, presence of ≥5 CA as compared to 3-4 CA increased the hazard ratio (HR) to 3.34 (P=0.017). When we limited the analysis to the subgroup of pts without evidence of a molecular TP53 mutation, presence of a cytogenetic TP53 deletion resulted in a HR of 5.67 (P=0.029). When the analysis was restricted to pts without a TP53 deletion, presence of a molecular TP53 mutation did not significantly change the HR (1.58, P=0.448; n.s.). Multivariate analysis that considered molecular TP53 mutation status, cytogenetic TP53 deletion status, the number of CA and treatment with azacitidine identified presence of a cytogenetic TP53 deletion as the most significant prognostic marker for OS (HR 15.1, P=0.010). HR was increased for pts with ≥5 CA compared to pts with 3-4 CA (HR 5.9, P=0.012). The molecular TP53 mutation status showed no significant impact on HR in our cohort. Conclusion: Presence of a cytogenetic TP53 deletion and a higher number of cytogenetic aberrations (≥5) showed a negative prognostic impact even within the unfavorable cytogenetic subgroup of MDS with complex abnormalities. In contrast, we found no strong prognostic impact for a molecular TP53 mutation in our cohort of MDS with complex abnormalities. The lower impact of TP53 mutations compared to TP53 deletions might be due to the lower degree of cytogenetic imbalances in pts with TP53 mutations only compared to pts with TP53 deletions only. However, molecular as well as cytogenetic TP53 aberrations were associated with a greater extent of chromosomal imbalances and displayed clear interdependencies. Our data suggest that TP53 alterations (molecular mutations and/or cytogenetic deletions) may not have an independent prognostic impact in the MDS subgroup with complex abnormalities. Figure 1 Figure 1. Figure 2 Figure 2. Figure 3 Figure 3. Disclosures Platzbecker: Amgen: Honoraria, Research Funding; TEVA Pharmaceutical Industries: Honoraria, Research Funding; Janssen-Cilag: Honoraria, Research Funding; Celgene Corporation: Honoraria, Research Funding; Novartis: Honoraria, Research Funding. Kroeger:Novartis: Honoraria, Research Funding.

The authors describe the case of a 63-year-old, female nursing home inhabitant suffering from trisomy 21, who accidentally ingested the anti-epileptic medication of another nursing home inhabitant. After telephone instructions from a specialist in internal medicine, caregivers forced the woman to vomit by means of saline solution and digital manipulation. This caused not only substantial hypernatriaemia but also aspiration pneumonia, from which the woman died after short hospitalization. The potential toxicity by major electrolyte shifts in terms of hypernatriaemia following administration of sodium chloride solution is well known; this measure is medically contraindicated for the induction of vomiting. The mechanisms leading to death in this case are presented, differentiated and discussed against the background of the literature.

Current techniques for the production of transgenic domestic animals remain inefficient. Only recently, DNA transposons resulted in improved efficiencies for mouse and pig transgenesis. In this work, we evaluated Tn5 and Sleeping Beauty systems for transgenesis in bovine and ovine species. First, both transposon systems were assessed in vitro in bovine embryos employing transposons carrying fluorescent reporter genes. In vitro-produced bovine zygotes were microinjected with either 1) a complex of Tn5:egfp transposon (20 ng μL–1) (protein: transgene with mosaic ends recognised by Tn5, in Mg+2 free medium), or 2) two plasmids carrying Sleeping Beauty 100X (pSB100X, 5 ng μL–1) and pT2/Venus transposon (10 ng μL–1). In vitro results for Tn5 transgenesis in bovine showed that blastocysts, Day 4 egfp embryos and egfp blastocysts rates for the group injected with Tn5:egfp did not differ from the group injected with the egfp transposon alone (73/145, 50%; 86/145, 59%; and 65/145, 45% v. 65/129, 50%; 87/129, 67%; and 57/129, 44%, respectively). For SB transgenesis, blastocysts, D4 Venus embryos, and Venus blastocysts rates did not differ between co-injection of pSB100X and pT2/Venus or injection with pT2/Venus alone (46/99, 46.5%; 64/99, 64.6%; and 33/99, 33.3% v. 41/83, 49.4%; 52/83, 62.7%; and 26/83, 31.3%, respectively). However, Venus intensity in blastocysts was markedly higher for the group co-injected with pSB100X and pT2/Venus respective to pT2/Venus alone. Both systems were assessed in vivo for the production of transgenic lambs employing a functional transposon (hrFIX, recombinant human factor IX driven by a Beta-lactoglobulin promoter). Laparoscopic artificial insemination of donor sheep was performed, and presumptive zygotes were flushed from the oviducts. The microinjections were done identically as described for the bovine embryos. A total of 24 presumptive zygotes were recovered and injected with the Tn5:hrFIX complex. Then, 21 zygotes were transferred to 5 synchronized ewes; one pregnancy of siblings was obtained, and one animal was born. Genomic DNA from skin, placenta, and blood was genotyped by PCR, but the hrFIX gene could not be detected. For the SB approach, 64 presumptive zygotes were recovered from 4 superovulated ewes, microinjected with the SB plasmids, and 21 of them were transferred to 7 oestrous synchronized recipients. The remaining zygotes were cultured in vitro and blastocysts (n = 7) were vitrified. Currently, 3 donor ewes are pregnant, one with siblings (4 total fetuses). Deliveries are expected by the end of August of this year. Our results indicate that both Tn5 and SB systems are capable of resulting in the production of transgene expressing embryos, and the presence of the transposases does not affect embryo viability. However, phenotyping of blastocyst stages does not seem to be predictive for stable transgene integration. The in vivo results will help to better address the suitability of Tn5 and SB approaches for the production of transgenic sheep.

The Champion West (CW) field Offshore Brunei, North Borneo consists of alarge number of vertically stacked hydrocarbon bearing reservoirs where thinlayers of laterally continuous shales are sealing the different hydrocarboncolumns. Wells in CW are typically completed selectively on multiple reservoirs (3-5zones) due to contrasting reservoir pressures and fluid properties and forreservoir management purposes. Although the reservoir rock is relativelyconsolidated, sand exclusion has been installed on shallower reservoirsfollowing sand failure in offset wells. Internal Gravel Packing (IGP) has inthe past been the preferred sand exclusion technique. Due to a high deviation in the four CW Early Oil Development (EOD) wells(54-66 deg.), it was desired to incorporate remote operated smart wellequipment to minimise future well intervention requirements. As the large distance between zones would require up to four IGP operationsper well (resulting in high costs) and would not provide sufficientthrough-bore to install the smart well equipment, it was decided to useExpandable Sand Screens (ESS) for sand control purposes in the new wells. Theuse of ESS facilitates multiple zones to be completed in single installationswhilst providing a larger through-bore for the installation of smart wellequipment. The paper discusses the sand-face completion design and installationaspects. As one of the EOD wells was completed with IGP, a benchmarking (rigtime, cost, performance) of the ESS technique is also presented. Finally, recommendations are made which should enable further cost reductions in the future.

Previous publications have produced conflicting results concerning the effect and nature of uterine horn position on embryonic weight and development in the uterus bicornis. For this investigation we used 503 embryos from 41 litters of NMRI - mice on gestational day 14. The relative position of each embryo was determined by measuring the distance from the center of each placenta to the cervix in relation to the total length of the uterine horn. After cessation of shrinkage due to fixation the umbilical cord and remaining membranes were removed under the dissecting microscope and the embryos were weighed to the nearest 0.1 mg. Analysis of potential embryonic positions along the uterine horns indicated that 10% at the extreme ovarian and cervical ends were almost devoid of implantations. A mixed nested analysis of variance was used as the appropriate model to test a position on weight effect. Not only was a position effect found but, in contrast to other studies, it is also quantitatively represented. Embryos were lighter both at the ovarian and cervical ends, the mean weight difference being in the range of less than 10% of the average embryo weight.