Knowledge skills and abilities demanded of graduates in the new learning environment

Changes in technology and employer demands require that we regularly survey our stakeholder communities to ensure the relevance and currency of what we are teaching, and the qualifications we offer to prepare our students for practice. At a presentation to an IT breakfast of the NZCS Auckland Branch in October 1998, the audience were surveyed in order to gauge how well we were doing in developing tomorrow's practitioners. Topics surveyed were: the relative importance of different technical skills required by employers, the key trends that we need to prepare graduates for, the impact of the Internet on skills required, the most urgent up skilling requirements of employees and the relative value of vendor vs. formal Polytechnic qualifications. The results of the survey are reported, the key findings analysed and some strategies are suggested which address the identified gaps

A western diet increases serotonin availability in rat small intestine

Diet-induced obesity is associated with changes in gastrointestinal function and induction of a mild inflammatory state. Serotonin (5-HT) containing enterochromaffin (EC) cells within the intestine respond to nutrients and are altered by inflammation. Thus, our aim was to characterize the uptake and release of 5-HT from EC cells of the rat ileum in a physiologically relevant model of diet-induced obesity. In chow-fed (CF) and Western diet-fed (WD) rats electrochemical methods were used to measure compression evoked (peak) and steady state (SS) 5-HT levels with fluoxetine used to block the serotonin reuptake transporter (SERT). The levels ofmRNAfor tryptophan hydroxylase 1 (TPH1) and SERT were determined by quantitative PCR, while EC cell numbers were determined immunohistochemically. In WD rats, the levels of 5-HT were significantly increased (SS: 19.2±3.7 ±M; peak: 73.5±14.1 ±M) compared with CF rats (SS: 12.3±1.8 ±M; peak: 32.2±7.2 ±M), while SERTdependent uptake of 5-HT was reduced (peak WD: 108% of control versus peak CF: 212% control). In WD rats, there was a significant increase in TPH1 mRNA, a decrease in SERT mRNA and protein, and an increase in EC cells. In conclusion, our data show that foods typical of a Western diet are associated with an increased 5-HT availability in the rat ileum. Increased 5-HT availability is driven by the up-regulation of 5-HT synthesis genes, decreased re-uptake of 5-HT, and increased numbers and/or 5-HT content of EC cells which are likely to cause altered intestinal motility and sensation in vivo. Copyright © 2010 The Endocrine Society. All rights reserved

Corporate Water Stewardship through Water Neutrality: A Case Study on the First Waterneutral® Apparel Factory in Asia

‘Water Neutrality’ is a novel concept first coined by the Water Footprint Network. ‘Water Neutral’ means that one reduces the water footprint of an activity as much as reasonably possible and offsets the negative externalities of the remaining water footprint. This does not indicate water usage being reduced to zero, but rather demonstrates that the negative economic, social and environmental externalities are reduced as much as possible, and that the remaining impacts are fully compensated. Compensation may be in the form of monetary contributions toward a more sustainable and equitable use of water within the hydrological system where the adverse impacts of the remaining “water footprint” are distributed. Linea Aqua, a subsidiary of the MAS holding, and one of the leading swimwear manufacturers in Asia, has taken a leading role in water stewardship to become Asia’s First Water Neutral® Apparel Factory. Linea Aqua’s water foot print was calculated annually (2013-2016) according to the methodologies developed by Water Footprint Network to quantify the impact to the water cycle of the surrounding ecosystem. Mitigation measures were introduced to reduce the identified impacts. According to the Water Footprint assessments, the total factory’s water intake has reduced by 5.9%. Daily water consumption per employee and per piece has reduced by 11.4% and 40.3% respectively when compared with the 2013 baseline. The factory has reduced the waste water discharge by 57%. Treated waste water reuse for flushing and gardening has increased by 39.8% when compared with the year 2014. Total 28% reduction of water intake was estimated with the installation of a Reverse Osmosis plant to recover waste water to reuse in cooling towers in the factory. Considering all the water savings, the total water recovery is 43.98% of the total water consumption. The balance saving of 32,990.12 m3/year has been achieved through the reasonable investment on pre- identified water replenishing projects on conservation or restoration of water quantity or quality within and outside of the affected water catchment. The overall reasonable investment for the replenish benefits as a function of cost share is Rs.14.00 million (2014- 2016).Keywords: Water Neutral, Water Footprint, Water stewardshi

Product Carbon Footprint of Wooden Products in Sri Lanka Special Reference to a Life Cycle of an Arm Chair

Forest and forest products have a vital role in mitigation of the global climate change. Themain objective of this study was to assess the carbon emissions in the manufacturing of woodproducts taking an example of the production of an arm chair using life cycle assessmentapproach (LCA) which provides a methodological framework for evaluating environmentalperformance over the life cycle of a product, process, or an activity.The product Carbon Footprint was assessed for a typical wooden arm chair manufactured inan average sawmill. Assessment boundary was cradle to grave. The system boundaryencompasses each product manufacturing process including material (logs, wood, resin,fuels) transport to each production facility. Transportation distances were reported in surveysand used to calculate product transported per kilogram–kilometers (kg-km). The embedded Cflux in harvested timber, GHG emissions and the flow of embedded CO2 stock during theprocessing were analysed. The cumulative system boundary includes all upstream flows ofenergy, fuel, and raw material for production. Energy consumed during transportationbetween the harvesting life-cycle stage and manufacturing accounts for actual distancesreported from each production region.The functional unit for the product was referenced to 1 m3 of the product. All input andoutput data within the cumulative system boundary were allocated to the functional unit ofproduct and co-products in accordance with International Organisation for Standardisation.The data represent average regional data from sample studies. Umberto for Carbon Softwarewas used for the analysis. The total life cycle consists of 08 stages, raw material and timberharvesting, timber depot operations, log conversions, wood preservation, timber seasoning,manufacturing, use phase, end of use/ final disposal.Based on the calculations, percentages of GHG emissions in each stage of life cycle were54% for raw material and timber harvesting, 4% for timber depot operations, 11% for logconversions, 5% wood preservation, 1% for timber seasoning, 12% for manufacturing and13% for use phase and end of use/final disposal. From the above results it is apparent that ascarbon emissions are greatest at the timber harvesting stage, measures should be taken tointroduce more efficient and effective machinery and methodologies to reduce the emissions.Keywords: Product carbon footprint, Wood products, Life cycle analysis, Climate chang

The human ankyrin 1 promoter insulator sustains gene expression in a β-globin lentiviral vector in hematopoietic stem cells.

Lentiviral vectors designed for the treatment of the hemoglobinopathies require the inclusion of regulatory and strong enhancer elements to achieve sufficient expression of the β-globin transgene. Despite the inclusion of these elements, the efficacy of these vectors may be limited by transgene silencing due to the genomic environment surrounding the integration site. Barrier insulators can be used to give more consistent expression and resist silencing even with lower vector copies. Here, the barrier activity of an insulator element from the human ankyrin-1 gene was analyzed in a lentiviral vector carrying an antisickling human β-globin gene. Inclusion of a single copy of the Ankyrin insulator did not affect viral titer, and improved the consistency of expression from the vector in murine erythroleukemia cells. The presence of the Ankyrin insulator element did not change transgene expression in human hematopoietic cells in short-term erythroid culture or in vivo in primary murine transplants. However, analysis in secondary recipients showed that the lentiviral vector with the Ankyrin element preserved transgene expression, whereas expression from the vector lacking the Ankyrin insulator decreased in secondary recipients. These studies demonstrate that the Ankyrin insulator may improve long-term β-globin expression in hematopoietic stem cells for gene therapy of hemoglobinopathies

The Macrocyclic Peptide Natural Product CJ-15,208 Is Orally Active and Prevents Reinstatement of Extinguished Cocaine-Seeking Behavior

The macrocyclic tetrapeptide natural product CJ-15,208 (cyclo[Phe-d-Pro-Phe-Trp]) exhibited both dose-dependent antinociception and kappa opioid receptor (KOR) antagonist activity after oral administration. CJ-15,208 antagonized a centrally administered KOR selective agonist, providing strong evidence it crosses the blood–brain barrier to reach KOR in the CNS. Orally administered CJ-15,208 also prevented both cocaine- and stress-induced reinstatement of extinguished cocaine-seeking behavior in the conditioned place preference assay in a time- and dose-dependent manner. Thus, CJ-15,208 is a promising lead compound with a unique activity profile for potential development, particularly as a therapeutic to prevent relapse to drug-seeking behavior in abstinent subjects

Creating New β-Globin-Expressing Lentiviral Vectors by High-Resolution Mapping of Locus Control Region Enhancer Sequences.

Hematopoietic stem cell gene therapy is a promising approach for treating disorders of the hematopoietic system. Identifying combinations of cis-regulatory elements that do not impede packaging or transduction efficiency when included in lentiviral vectors has proven challenging. In this study, we deploy LV-MPRA (lentiviral vector-based, massively parallel reporter assay), an approach that simultaneously analyzes thousands of synthetic DNA fragments in parallel to identify sequence-intrinsic and lineage-specific enhancer function at near-base-pair resolution. We demonstrate the power of LV-MPRA in elucidating the boundaries of previously unknown intrinsic enhancer sequences of the human β-globin locus control region. Our approach facilitated the rapid assembly of novel therapeutic βAS3-globin lentiviral vectors harboring strong lineage-specific recombinant control elements capable of correcting a mouse model of sickle cell disease. LV-MPRA can be used to map any genomic locus for enhancer activity and facilitates the rapid development of therapeutic vectors for treating disorders of the hematopoietic system or other specific tissues and cell types

Relaxin deficiency leads to uterine artery dysfunction during pregnancy in mice

The uterine vasculature undergoes profound adaptations in response to pregnancy. Augmentation of endothelial vasodilator function and reduced smooth muscle reactivity are factors contributing to uterine artery adaptation and are critical for adequate placental perfusion. The peptide hormone relaxin has an important role in mediating the normal maternal renal vascular adaptations during pregnancy through a reduction in myogenic tone and an increase in flow-mediated vasodilation. Little is known however about the influence of endogenous relaxin on the uterine artery during pregnancy. We tested the hypothesis that relaxin deficiency increases myogenic tone and impairs endothelial vasodilator function in uterine arteries of late pregnant relaxin deficient (Rln−/−) mice. Reactivity of main uterine arteries from non-pregnant and late pregnant wild-type (Rln+/+) and Rln−/− mice was studied using pressure and wire myography and changes in gene expression explored using PCR. Myogenic tone was indistinguishable in arteries from non-pregnant mice. In late pregnancy uterine artery myogenic tone was halved in Rln+/+ mice (P < 0.0001), an adaptation that failed to occur in arteries from pregnant Rln−/− mice. The role of vasodilator prostanoids in the regulation of myogenic tone was significantly reduced in arteries of pregnant Rln−/− mice (P = 0.02). Agonist-mediated endothelium-dependent vasodilation was significantly impaired in non-pregnant Rln−/− mice. With pregnancy, differences in total endothelial vasodilator function were resolved, although there remained an underlying deficiency in the role of vasodilator prostanoids and alterations to the contributions of calcium-activated K+ channels. Fetuses of late pregnant Rln−/− mice were ~10% lighter (P < 0.001) than those of Rln+/+ mice. In conclusion, relaxin deficiency is associated with failed suppression of uterine artery myogenic tone in pregnancy, which likely contributes to reduced uteroplacental perfusion and fetal growth restriction.Sarah A. Marshall, Sevvandi N. Senadheera, Maria Jelinic, Kelly O’Sullivan, Laura J. Parry and Marianne Tar

Phenylalanine Stereoisomers of CJ-15,208 and [d-Trp]CJ-15,208 Exhibit Distinctly Different Opioid Activity Profiles

This work is licensed under a Creative Commons Attribution 4.0 International License.The macrocyclic tetrapeptide cyclo[Phe-d-Pro-Phe-Trp] (CJ-15,208) and its stereoisomer cyclo[Phe-d-Pro-Phe-d-Trp] exhibit different opioid activity profiles in vivo. The present study evaluated the influence of the Phe residues’ stereochemistry on the peptides’ opioid activity. Five stereoisomers were synthesized by a combination of solid-phase peptide synthesis and cyclization in solution. The analogs were evaluated in vitro for opioid receptor affinity in radioligand competition binding assays, and for opioid activity and selectivity in vivo in the mouse 55 °C warm-water tail-withdrawal assay. Potential liabilities of locomotor impairment, respiratory depression, acute tolerance development, and place conditioning were also assessed in vivo. All of the stereoisomers exhibited antinociception following either intracerebroventricular or oral administration differentially mediated by multiple opioid receptors, with kappa opioid receptor (KOR) activity contributing for all of the peptides. However, unlike the parent peptides, KOR antagonism was exhibited by only one stereoisomer, while another isomer produced DOR antagonism. The stereoisomers of CJ-15,208 lacked significant respiratory effects, while the [d-Trp]CJ-15,208 stereoisomers did not elicit antinociceptive tolerance. Two isomers, cyclo[d-Phe-d-Pro-d-Phe-Trp] (3) and cyclo[Phe-d-Pro-d-Phe-d-Trp] (5), did not elicit either preference or aversion in a conditioned place preference assay. Collectively, these stereoisomers represent new lead compounds for further investigation in the development of safer opioid analgesics.National Institute on Drug Abuse (R01 DA18832)National Institute on Drug Abuse (R01 DA032928

Preclinical Demonstration of Lentiviral Vector-mediated Correction of Immunological and Metabolic Abnormalities in Models of Adenosine Deaminase Deficiency

Gene transfer into autologous hematopoietic stem cells by γ-retroviral vectors (gRV) is an effective treatment for adenosine deaminase (ADA)–deficient severe combined immunodeficiency (SCID). However, current gRV have significant potential for insertional mutagenesis as reported in clinical trials for other primary immunodeficiencies. To improve the efficacy and safety of ADA-SCID gene therapy (GT), we generated a self-inactivating lentiviral vector (LV) with a codon-optimized human cADA gene under the control of the short form elongation factor-1α promoter (LV EFS ADA). In ADA−/− mice, LV EFS ADA displayed high-efficiency gene transfer and sufficient ADA expression to rescue ADA−/− mice from their lethal phenotype with good thymic and peripheral T- and B-cell reconstitution. Human ADA-deficient CD34+ cells transduced with 1–5 × 107 TU/ml had 1–3 vector copies/cell and expressed 1–2x of normal endogenous levels of ADA, as assayed in vitro and by transplantation into immune-deficient mice. Importantly, in vitro immortalization assays demonstrated that LV EFS ADA had significantly less transformation potential compared to gRV vectors, and vector integration-site analysis by nrLAM-PCR of transduced human cells grown in immune-deficient mice showed no evidence of clonal skewing. These data demonstrated that the LV EFS ADA vector can effectively transfer the human ADA cDNA and promote immune and metabolic recovery, while reducing the potential for vector-mediated insertional mutagenesis