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Collaboration between research and policy is an essential element for knowledge-based public health. However, only half of the Danish municipalities have experience with collaborating with researchers or other stakeholders. Through content analysis of interviews and policy documents the study explores the involvement of external stakeholders in local health policymaking and public officials' perceptions on involving them. Main involvement was through a personal contact or through a regular hearing. The purpose of involvement was mostly tactical or to solve problems. Politicians had substantial influence on the involvement of external stakeholders, allowing only a few to contribute in a closed policymaking process.

Phenylalanine hydroxylase (PAH) is a crucial enzyme involved in tyrosine biosynthesis, having roles in neurological and physiological processes. The purpose of PAH has received little attention in crustaceans despite extensive investigations in other arthropods. Here, we characterize the PAH gene for the first time in the parasite Lepeophtheirus salmonis, a copepod that is responsible for huge economic losses in salmonid fish farming. Phylogenetic and sequence analyses confirmed that LsPAH is closely related to the metazoan PAH with conserved ACT regulatory and catalytic domains. Temporal expression patterns revealed that LsPAH is expressed throughout all developmental stages peaking during the copepodite stages, suggesting an essential role in developmental physiology. We used RNAi to knockdown LsPAH expression in the nauplius I stage to study developmental function during the larval stages. PAH knockdown impaired larval development, molting and swimming ability with severe morphological defects. This study provides insight into the role of PAH in copepods and demonstrates the importance of this metabolic gene in salmon louse growth and development.

Objective Obesity is an increasing public health challenge and most weight loss programmes are still inadequate to support sustainable weight loss. One reason for the continued lack of success might be the dominant biomedical, individualised approach to weight loss. Holistic approaches that focus on overall health and well-being in addition to weight loss are increasingly recommended. In Denmark, health professionals in the municipalities are responsible for developing and conducting weight loss programmes. The objective of this study was to explore what health professional’s perceived as an ideal, holistic weight loss programme that could be feasibly implemented in the municipalities.Design A phenomenological-hermeneutical qualitative study was performed using semistructured interviews.Setting Thirty-two Danish municipalities were weight loss programmes are developed and conducted.Participants Thirty-five health professionals with experience conducting weight loss programmes.Results Three themes emerged from the analysis: Support from the social network are important both during and after a weight loss, Changing the self-belief by positive discussions and doing activities, Maintaining changes through daily life.Conclusion Future municipal weight loss programmes should emphasise overall health and well-being instead of weight loss and adopt a holistic approach including a focus on social relationships, meaningful activities and successes as part of a balanced daily life.

Several mutations in αB-crystallin (CryAB), a heat shock protein with chaperone-like activities, are causally linked to skeletal and cardiac myopathies in humans. To better understand the underlying pathogenic mechanisms, we had previously generated transgenic (TG) mice expressing R120GCryAB, which recapitulated distinguishing features of the myopathic disorder (e.g., protein aggregates, hypertrophic cardiomyopathy). To determine whether induced pluripotent stem cell (iPSC)-derived cardiomyocytes, a new experimental approach for human disease modeling, would be relevant to aggregation-prone disorders, we decided to exploit the existing transgenic mouse model to derive iPSCs from tail tip fibroblasts. Several iPSC lines were generated from TG and non-TG mice and validated for pluripotency. TG iPSC-derived cardiomyocytes contained perinuclear aggregates positive for CryAB staining, whereas CryAB protein accumulated in both detergent-soluble and insoluble fractions. iPSC-derived cardiomyocytes identified by cardiac troponin T staining were significantly larger when expressing R120GCryAB at a high level in comparison with TG low expressor or non-TG cells. Expression of fetal genes such as atrial natriuretic factor, B-type natriuretic peptide, and α-skeletal α-actin, assessed by quantitative reverse transcription-polymerase chain reaction, were increased in TG cardiomyocytes compared with non-TG, indicating the activation of the hypertrophic genetic program in vitro. Our study demonstrates for the first time that differentiation of R120G iPSCs into cardiomyocytes causes protein aggregation and cellular hypertrophy, recapitulating in vitro key pathognomonic hallmarks found in both animal models and patients. Our findings pave the way for further studies exploiting this cell model system for mechanistic and therapeutic investigations.

Intertidal benthic diatoms experience a highly variable light regime, which especially challenges these organisms to cope with excess light energy during low tide. Non-photochemical quenching of chlorophyll fluorescence (NPQ) is one of the most rapid mechanisms diatoms possess to dissipate excess energy. Its capacity is mainly defined by the xanthophyll cycle (XC) and Light-Harvesting Complex X (LHCX) proteins. Whereas the XC and its relation to NPQ have been relatively well-studied in both planktonic and benthic diatoms, our current knowledge about LHCX proteins and their potential involvement in NPQ regulation is largely restricted to planktonic diatoms. While recent studies using immuno-blotting have revealed the presence of light regulated LHCX proteins in benthic diatom communities and isolates, nothing is as yet known about the diversity, identity and transcriptional regulation or function of these proteins. We identified LHCX genes in the draft genome of the model benthic diatom Seminavis robusta and followed their transcriptional regulation during a day/night cycle and during exposure to high light conditions. The S. robusta genome contains 17 LHCX sequences, which is much more than in the sequenced planktonic model diatoms (4–5), but similar to the number of LHCX genes in the sea ice associated diatom Fragilariopsis cylindrus. LHCX diversification in both species, however, appears to have occurred independently. Interestingly, the S. robusta genome contains LHCX genes that are related to the LHCX6 of the model centric diatom Thalassiosira pseudonana, which are lacking in the well-studied pennate model diatom Phaeodactylum tricornutum. All investigated LHCX genes, with exception of SrLHCX6, were upregulated during the daily dark-light transition. Exposure to 2,000 μmol photons m–2 s–1, furthermore, increased transcription of all investigated LHCX genes. Our data suggest that the diversification and involvement of several light regulated LHCX genes in the photophysiology of S. robusta may represent an adaptation to the complex and highly variable light environment this benthic diatom species can be exposed to.

Glucosinolates are defense-related secondary metabolites found in Brassicaceae. When Brassicaceae come under attack, glucosinolates are hydrolyzed into different forms of glucosinolate hydrolysis products (GHPs). Among the GHPs, isothiocyanates are the most comprehensively characterized defensive compounds, whereas the functional study of nitriles, another group of GHP, is still limited. Therefore, this study investigates whether 3-butenenitrile (3BN), a nitrile, can trigger the signaling pathways involved in the regulation of defense responses in Arabidopsis thaliana against biotic stresses. Briefly, the methodology is divided into three stages, (i) evaluate the physiological and biochemical effects of exogenous 3BN treatment on Arabidopsis, (ii) determine the metabolites involved in 3BN-mediated defense responses in Arabidopsis, and (iii) assess whether a 3BN treatment can enhance the disease tolerance of Arabidopsis against necrotrophic pathogens. As a result, a 2.5 mM 3BN treatment caused lesion formation in Arabidopsis Columbia (Col-0) plants, a process found to be modulated by nitric oxide (NO). Metabolite profiling revealed an increased production of soluble sugars, Krebs cycle associated carboxylic acids and amino acids in Arabidopsis upon a 2.5 mM 3BN treatment, presumably via NO action. Primary metabolites such as sugars and amino acids are known to be crucial components in modulating plant defense responses. Furthermore, exposure to 2.0 mM 3BN treatment began to increase the production of salicylic acid (SA) and jasmonic acid (JA) phytohormones in Arabidopsis Col-0 plants in the absence of lesion formation. The production of SA and JA in nitrate reductase loss-of function mutant (nia1nia2) plants was also induced by the 3BN treatments, with a greater induction for JA. The SA concentration in nia1nia2 plants was lower than in Col-0 plants, confirming the previously reported role of NO in controlling SA production in Arabidopsis. A 2.0 mM 3BN treatment prior to pathogen assays effectively alleviated the leaf lesion symptom of Arabidopsis Col-0 plants caused by Pectobacterium carotovorum ssp. carotovorum and Botrytis cinerea and reduced the pathogen growth on leaves. The findings of this study demonstrate that 3BN can elicit defense response pathways in Arabidopsis, which potentially involves a coordinated crosstalk between NO and phytohormone signaling.

Background. Individuals with Parkinson’s Disease (PD) have bradykinesia during mobility tasks in the morning before intake of dopaminergic treatment and have difficulties managing Activities of Daily Living (ADLs). Early morning off (EMO) refers to off-states in the morning where the severity of bradykinesia is increased and causes a decrease in mobility related to wearing off of effects of medication. Measurements from devices capable of continuously recording motor symptoms may provide insight into the patient’s response to medication and possible impact on ADLs. Objectives. To test whether poor or slow response to medication in the morning predicts the overall ADL-level and to assess the association between change in bradykinesia score (BKS) and the risk of having disabilities within three selected ADL-items. Methods. In this cross-sectional study, the sample consists of 34 patients with light to moderate PD. Data collection encompasses measurements from the Parkinson KinetiGraph, and the ADL-limitations are assessed by the Movement Disorder Society Unified Parkinson Disease Rating Scale (MDS-UPDRS) Part II. Results. The association between UPDRS- II and BKS from the algorithm was −0.082 (p

The roles of host-associated bacteria have gained attention lately, and we now recognise that the microbiota is essential in processes such as digestion, development of the immune system and gut function. In this study, Atlantic cod larvae were reared under germ-free, gnotobiotic and conventional conditions. Water and fish microbiota were characterised by 16S rRNA gene analyses. The cod larvae’s transcriptional responses to the different microbial conditions were analysed by a custom Agilent 44 k oligo microarray. Gut development was assessed by transmission electron microscopy (TEM). Water and fish microbiota differed significantly in the conventional treatment and were dominated by different fast-growing bacteria. Our study indicates that components of the innate immune system of cod larvae are downregulated by the presence of non-pathogenic bacteria, and thus may be turned on by default in the early larval stages. We see indications of decreased nutrient uptake in the absence of bacteria. The bacteria also influence the gut morphology, reflected in shorter microvilli with higher density in the conventional larvae than in the germ-free larvae. The fact that the microbiota alters innate immune responses and gut morphology demonstrates its important role in marine larval development.

Parkinson's Disease (PD) and Multiple System Atrophy (MSA) are neurodegenerative diseases characterized neuropathologically by alpha-synuclein accumulation in brain cells. This accumulation is hypothesized to contribute to constitutive neuroinflammation, and to participate in the neurodegeneration. Cytokines, which are the main inflammatory signalling molecules, have been identified in blood and cerebrospinal fluid of PD patients, but studies investigating the human brain levels are scarce. It is documented that neurotrophins, necessary for survival of brain cells and known to interact with cytokines, are altered in the basal ganglia of PD patients. In regards to MSA, no major study has investigated brain cytokine or neurotrophin protein expression.Here, we measured protein levels of 18 cytokines (IL-2, 4–8, 10, 12, 13, 17, G-CSF, GM-CSF, IFN-γ, MCP-1, MIP-1α and 1β, TNF-α) and 5 neurotrophins (BDNF, GDNF, bFGF, PDGF-BB, VEGF) in the dorsomedial prefrontal cortex in brains of MSA and PD patients and control subjects. We found altered expression of IL-2, IL-13, and G-CSF, but no differences in neurotrophin levels. Further, in MSA patients we identified increased mRNA levels of GSK3β that is involved in neuroinflammatory pathways. Lastly, we identified increased expression of the neurodegenerative marker S100B, but not CRP, in PD and MSA patients, indicating local rather than systemic inflammation. Supporting this, in both diseases we observed increased MHC class II+ and CD45+ positive cells, and low numbers of infiltrating CD3+ cells. In conclusion, we identified neuroinflammatory responses in PD and MSA which seems more widespread in the brain than neurotrophic changes.

Abstract The diatom cell wall, or frustule, is a highly complex, three-dimensional structure consisting of nanopatterned silica as well as proteins and other organic components. While some key components have been identified, knowledge on frustule biosynthesis is still fragmented. The model diatom Thalassiosira pseudonana was subjected to silicon (Si) shift-up and shift-down situations. Cellular and molecular signatures, dynamic changes and co-regulated clusters representing the hallmarks of cellular and molecular responses to changing Si availabilities were characterised. Ten new proteins with silaffin-like motifs, two kinases and a novel family of putatively frustule-associated transmembrane proteins induced by Si shift-up with a possible role in frustule biosynthesis were identified. A separate cluster analysis performed on all significantly regulated silaffin-like proteins (SFLPs), as well as silaffin-like motifs, resulted in the classification of silaffins, cingulins and SFLPs into distinct clusters. A majority of the genes in the Si-responsive clusters are highly divergent, but positive selection does not seem to be the driver behind this variability. This study provides a high-resolution map over transcriptional responses to changes in Si availability in T. pseudonana. Hallmark Si-responsive genes are identified, characteristic motifs and domains are classified, and taxonomic and evolutionary implications outlined and discussed.

Abstract Background Germline mutations in genes encoding subunits of succinate dehydrogenase (SDH) are associated with the development of pheochromocytoma (PC) and/or paraganglioma (PGL). As assembly factors have been identified as playing a role in maturation of individual SDH subunits and assembly of the functioning SDH complex, we hypothesized that SDHAF3 variants may be associated with PC/PGL and functionality of SDH. Methods DNA was extracted from the blood of 37 individuals (from 23 families) with germline SDH mutations and 18 PC/PGL (15 sporadic, 3 familial) and screened for mutations using a custom gene panel, containing SDHAF3 (SDH assembly factor 3) as well as eight known PC/PGL susceptibility genes. Molecular and functional consequences of an identified sequence variant of SDHAF3 were assessed in yeast and mammalian cells (HEK293). Results Using massively parallel sequencing, we identified a variant in SDHAF3, c.157 T > C (p.Phe53Leu), associated with increased prevalence in familial and sporadic PC/PGL (6.6%) when compared to normal populations (1.2% [1000 Genomes], p = 0.003; 2.1% [Exome Aggregation Consortium], p = 0.0063). In silico prediction tools suggest this variant is probably damaging to protein function, hence we assessed molecular and functional consequences of the resulting amino acid change (p.Phe53Leu) in yeast and human cells. We showed that introduction of SDHAF3 p.Phe53Leu into Sdh7 (ortholog of SDHAF3 in humans) null yeast resulted in impaired function, as observed by its failure to restore SDH activity when expressed in Sdh7 null yeast relative to WT SDHAF3. As SDHAF3 is involved in maturation of SDHB, we tested the functional impact of SDHAF3 c.157 T > C and various clinically relevant SDHB mutations on this interaction. Our in vitro studies in human cells show that SDHAF3 interacts with SDHB (residues 46 and 242), with impaired interaction observed in the presence of the SDHAF3 c.157 T > C variant. Conclusions Our studies reveal novel insights into the biogenesis of SDH, uncovering a vital interaction between SDHAF3 and SDHB. We have shown that SDHAF3 interacts directly with SDHB (residue 242 being key to this interaction), and that a variant in SDHAF3 (c.157 T > C [p.Phe53Leu]) may be more prevalent in individuals with PC/PGL, and is hypomorphic via impaired interaction with SDHB.

Abstract Background Parkinson’s’ disease (PD) and Multiple System Atrophy (MSA) are progressive brain disorders characterized by intracellular accumulations of α-synuclein and nerve cell loss in specific brain areas. This loss causes problems with movement, balance and/or autonomic functions. Naturally occurring autoantibodies (NAbs) play potentially an important role in clearing or/and blocking circulating pathological proteins. Little is known about the functional properties of anti-α-synuclein NAbs in PD and MSA, and there have been opposing reports regarding their plasma concentrations in these disorders. Methods We have investigated the apparent affinity of anti-α-synuclein NAbs in plasma samples from 46 PD patients, 18 MSA patients and 41 controls using competitive enzyme-linked immunosorbent assay (ELISA) and Meso Scale Discovery (MSD) set-ups. Results We found that the occurrence of high affinity anti-α-synuclein NAbs in plasma from PD patients is reduced compared to healthy controls, and nearly absent in plasma from MSA patients. Also, levels of α-synuclein/NAbs immunocomplexes is substantially reduced in plasma from both patient groups. Further, cross binding of anti-α-synuclein NAbs with β- and γ-synuclein monomers suggest, the high affinity anti-α-synuclein plasma component, seen in healthy individuals, is directed mainly against C-terminal epitopes. Furthermore, we also observed reduced occurrence of high affinity anti-phosphorylated-α-synuclein NAbs in plasma from PD and MSA patients. Conclusions One interpretation implies that these patients may have impaired ability to clear and/or block the effects of pathological α-synuclein due to insufficient/absent concentration of NAbs and as such provides a rationale for testing immune-based therapeutic strategies directed against pathological α-synuclein. Following this interpretation, we can hypothesize that high affinity autoantibodies efficiently bind and clear potentially pathological species of α-synuclein in healthy brain, and that this mechanism is impaired or absent in PD and MSA patients.

Abstract Background A limited amount of research has examined the effect of changing public open spaces on active living. This paper will present the study protocol of a community-based intervention study co-designed in an interdisciplinary collaboration with community members to develop urban installations highly tailored to promote active living among children (10–13-years-old) and seniors (>60-years-old) in a deprived neighbourhood in Copenhagen. Methods The study builds on a quasi-experimental study design with two sub-studies: 1) a children study and 2) a senior study. The interventions will be developed, designed and implemented in collaboration with local children and seniors, respectively, using different co-design tools and methods. We will evaluate the effect of the interventions on children’s and senior’s use of the new-built urban installations using accelerometers in combination with GPS as well as systematic observation using the System for Observing Play and Recreation in Communities (SOPARC). A process evaluation with focus groups consisting of the various stakeholders in the two sub-studies will be used to gain knowledge of the intervention processes. Discussion The paper presents new approaches in the field of public open space interventions through interdisciplinary collaboration, participatory co-design approach and combination of measurements. Using both effect and process evaluations the study will provide unique insights in the role and importance of the interdisciplinary collaboration, participatory processes, and tailoring changes in public open space to local needs and wishes. These results can be used to guide urban renewal projects in deprived neighbourhoods in the future. Trial registration Retrospectively registered with study ID ISRCTN50036837 . Date of registration: 16 December 2016.

The objective of the study was to find an appropriate Balanced Scorecard model to be implemented in the vocational education management and to test the effectiveness of balanced scorecard internal model for the vocational high school education. The method that the researchers implemented in the study was the research and development designed by Borg & Gall. The data in the study were gathered by means of in-depth interview, observation and documentary analysis in the five prominent vocational high schools from the Province of Southeast Sulawesi. The informants in the study were selected by means of snowball sampling. The data validity was tested by the triangulation technique. The First Stage and the Second Stage of model validation/ experiment was implemented by means of Delphi technique and the validation/experiment of both stages involved 50 leaders from the state and the private vocational high schools in the Province of Southeast Sulawesi. The study showed that the appropriate Balanced Scorecard model for the vocational high school educational management has been a system that might be implemented for identifying and for measuring the main performance of vocational high schools through four perspectives namely the financial perspective, the customer perspective, the internal process perspective and the learning and growth perspective. Balanced Scorecard model for the vocational high school education has met the criteria of quite high internal effectiveness model and has belonged to the good criteria. As a result, the Balanced Scorecard model might be implemented as the model of vocational high school managerial implementation. Keyword: balanced scorecard, vocational high school and vocational high school performance PENGEMBANGAN MODEL BALANCED SCORECARD UNTUK PENDIDIKAN SMK Abstrak Tujuan penelitian ini adalah mengembangkan model Balanced Scorecard yang cocok untuk diterapkan dalam pengelolaan pendidikan kejuruan dan menguji keefektifan internal model Balanced Scorecard untuk pendidikan SMK. Metode penelitian yang digunakan dalam penelitian ini adalah penelitian dan pengembangan (R&D) rancangan Borg & Gall. Data penelitian dikumpulkan melalui wawancara mendalam, observasi, dan analisis dokumen pada lima SMK Unggul di Provinsi Sulawesi Tenggara. Informan penelitian ditetapkan dengan snowball sampling. Keabsahan data diuji dengan teknik trianggulasi. Uji coba/validasi model Tahap 1 dan Tahap 2 menggunakan teknik Delphi dengan subjek penelitian sebanyak 50 orang pimpinan SMK dari 15 SMK Negeri dan SMK Swasta di Provinsi Sulawesi Tenggara. Penelitian ini menunjukkan bahwa model Balanced Scorecard yang cocok untuk mengelola manajemen SMK adalah suatu sistem yang digunakan untuk mengenali dan mengukur kinerja utama SMK melalui empat perspektif Balanced Scorecard meliputi perspektif keuangan, pelanggan, proses internal, pembelajaran dan pertumbuhan. Model Balanced Scorecard yang dikembangkan untuk pendidikan SMK yang diajukan telah memenuhi kriteria keefektifan internal model yang tinggi dan termasuk kategori sangat baik, sehingga layak digunakan sebagai model pelaksanaan manajemen SMK. Kata kunci: balanced scorecard, SMK, dan kinerja SMK

Aggregation of alpha-synuclein (α-syn) is considered to be the major pathological hallmark and driving force of Multiple System Atrophy (MSA) and Parkinson's disease (PD). Immune dysfunctions have been associated with both MSA and PD and recently we reported that the levels of natural occurring autoantibodies (NAbs) with high-affinity/avidity toward α-synuclein are reduced in MSA and PD patients. Here, we aimed to evaluate the plasma immunoglobulin (Ig) composition binding α-syn and other amyloidogenic neuropathological proteins, and to correlate them with disease severity and duration in MSA and PD patients. All participants were recruited from a single neurological unit and the plasma samples were stored for later research at the Bispebjerg Movement Disorder Biobank. All patients were diagnosed according to current consensus criteria. Using multiple variable linear regression analyses, we observed higher levels of anti-α-syn IgG1 and IgG3 NAbs in MSA vs. PD, higher levels of anti-α-syn IgG2 NAbs in PD compared to controls, whereas anti-α-syn IgG4 NAbs were reduced in PD compared to MSA and controls. Anti-α-syn IgM levels were decreased in both MSA and PD. Further our data supported that MSA patients' immune system was affected with reduced IgG1 and IgM global levels compared to PD and controls, with further reduced global IgG2 levels compared to PD. These results suggest distinct autoimmune patterns in MSA and PD. These findings suggest a specific autoimmune physiological mechanism involving responses toward α-syn, differing in neurodegenerative disease with overlapping α-syn pathology.

ABSTRACT: Samples of 168 maté trees (Ilex paraguariensis) from four natural populations of different states of Brazil (Rio Grande do Sul, Santa Catarina, Paraná and Mato Grosso do Sul) were analyzed for their variability in storage proteins seed, which were used to estimate the degree of differentiation among and within species, because are relatively stable during evolution. Data on band presence/absence from 80 different polypeptides were used to perform similarity coefficient of Jaccard. A high level of genetic variability was reported within sampled populations (SJ intra=0.374). Lower levels of diversity were observed between populations (SJ inter=0.308). Total genetic diversity of maté was Hsp=0.264 estimated by Shannon measure. Partition of that value disclosed that 95 % of the total diversity is within-population, and only 5% to between-populations. The phenogram based on King’s distances indicates a certain degree of geographic differentiation. This represents the first study on storage protein variability in I. paraguariensis and guides the choice of the specimens to increment germoplasm banks and genetic improvement programs.

An 8 month monitoring campaign on the Seyfert 1 galaxy Fairall 9 has been conducted with the International Ultraviolet Explorer in an attempt to obtain reliable estimates of continuum-continuum and continuum-emission-line delays for a high-luminosity active galactic nucleus (AGN). While the results of this campaign are more ambiguous than those of previous monitoring campaigns on lower luminosity sources, we find general agreement with the earlier results: (1) there is no measurable lag between ultraviolet continuum bands, and (2) the measured emission-line time lags are very short. It is especially notable that the Lyα + N v emission-line lag is about 1 order of magnitude smaller than determined from a previous campaign by Clavel, Wamsteker, & Glass (1989) when Fairall 9 was in a more luminous state. In other well-monitored sources, specifically NGC 5548 and NGC 3783, the highest ionization lines are found to respond to continuum variations more rapidly than the lower ionization lines, which suggests a radially ionization-stratified broad-line region. In this case, the results are less certain, since none of the emission-line lags are very well determined. The best-determined emission line lag is Lyα + N v, for which we find that the centroid of the continuum-emission-line cross-correlation function is τcent ≈ 14-20 days. We measure a lag τcent ≲ 4 days for He II λ1640; this result is consistent with the ionization-stratification pattern seen in lower luminosity sources, but the relatively large uncertainties in the emission-line lags measured here cannot rule out similar lags for Lyα + N v and He II λ1640 at a high level of significance. We are unable to determine a reliable lag for C IV λ1550, but we note that the profiles of the variable parts of Lyα and C IV λ1550 are not the same, which does not support the hypothesis that the strongest variations in these two lines arise in the same region.

Cox23 is a known conserved assembly factor for cytochrome c oxidase, although its role in cytochrome c oxidase (CcO) biogenesis remains unresolved. To gain additional insights into its role, we isolated spontaneous suppressors of the respiratory growth defect in cox23∆ yeast cells. We recovered independent colonies that propagated on glycerol/lactate medium for cox23∆ cells at 37°C. We mapped these mutations to the mitochondrial genome and specifically to COX1 yielding an I101F substitution. The I101F Cox1 allele is a gain-of-function mutation enabling yeast to respire in the absence of Cox23. CcO subunit steady-state levels were restored with the I101F Cox1 suppressor mutation and oxygen consumption and CcO activity were likewise restored. Cells harboring the mitochondrial genome encoding I101F Cox1 were used to delete genes for other CcO assembly factors to test the specificity of the Cox1 mutation as a suppressor of cox23∆ cells. The Cox1 mutant allele fails to support respiratory growth in yeast lacking Cox17, Cox19, Coa1, Coa2, Cox14 or Shy1, demonstrating its specific suppressor activity for cox23∆ cells.

Jacob Hyllested-Winge,1 Thomas Sparre,2 Line Kynemund Pedersen2 1Novo Nordisk Pharma Ltd, Tokyo, Japan; 2Novo Nordisk A/S, Søborg, Denmark Abstract: The introduction of insulin pen devices has provided easier, well-tolerated, and more convenient treatment regimens for patients with diabetes mellitus. When compared with vial and syringe regimens, insulin pens offer a greater clinical efficacy, improved quality of life, and increased dosing accuracy, particularly at low doses. The portable and discreet nature of pen devices reduces the burden on the patient, facilitates adherence, and subsequently contributes to the improvement in glycemic control. NovoPen Echo® is one of the latest members of the NovoPen® family that has been specifically designed for the pediatric population and is the first to combine half-unit increment (=0.5 U of insulin) dosing with a simple memory function. The half-unit increment dosing amendments and accurate injection of 0.5 U of insulin are particularly beneficial for children (and insulin-sensitive adults/elders), who often require small insulin doses. The memory function can be used to record the time and amount of the last dose, reducing the fear of double dosing or missing a dose. The memory function also provides parents with extra confidence and security that their child is taking insulin at the correct doses and times. NovoPen Echo is a lightweight, durable insulin delivery pen; it is available in two different colors, which may help to distinguish between different types of insulin, providing more confidence for both users and caregivers. Studies have demonstrated a high level of patient satisfaction, with 80% of users preferring NovoPen Echo to other pediatric insulin pens. Keywords: NovoPen Echo®, memory function, half-unit increment dosing, adherence, children, adolescents

An amendment to this paper has been published and can be accessed via a link at the top of the paper.