Development and characterization of a potent free fatty acid receptor 1 (FFA1) fluorescent tracer

The free fatty acid receptor 1 (FFA1/GPR40) is a potential target for treatment of type 2 diabetes. Although several potent agonists have been described, there remains a strong need for suitable tracers to interrogate ligand binding to this receptor. We address this by exploring fluorophore-tethering to known potent FFA1 agonists. This led to the development of 4, a high affinity FFA1 tracer with favorable and polarity-dependent fluorescent properties. A close to ideal overlap between the emission spectrum of the NanoLuciferase receptor tag and the excitation spectrum of 4 enabled the establishment of a homogenous BRET-based binding assay suitable for both detailed kinetic studies and high throughput competition binding studies. Using 4 as a tracer demonstrated that the compound acts fully competitively with selected synthetic agonists but not with lauric acid and allowed for the characterization of binding affinities of a diverse selection of known FFA1 agonists, indicating that 4 will be a valuable tool for future studies at FFA1

Development and characterization of a fluorescent tracer for the free fatty acid receptor 2 (FFA2/GPR43)

The free fatty acid receptor 2 (FFA2/GPR43) is considered a potential target for treatment of metabolic and inflammatory diseases. Here we describe the development of the first fluorescent tracer for FFA2 intended as a tool for assessment of thermodynamic and kinetic binding parameters of unlabeled ligands. Starting with a known azetidine FFA2 antagonist, we used a carboxylic acid moiety known not to be critical for receptor interaction as attachment point for a nitrobenzoxadiazole (NBD) fluorophore. This led to the development of 4 (TUG-1609), a fluorescent tracer for FFA2 with favorable spectroscopic properties and high affinity, as determined by bioluminescence resonance energy transfer (BRET)-based saturation and kinetic binding experiments, as well as a high specific to nonspecific BRET binding signal. A BRET-based competition binding assay with 4 was also established and used to determine binding constants and kinetics of unlabeled ligands

HUBUNGAN ANTARA KOMITMEN BERPACARAN DENGAN KUALITAS PERSAHABATAN PADA REMAJA AKHIR DI UNIVERSITAS PENDIDIKAN INDONESIA

Penelitian ini memiliki beberapa tujuan, yaitu untuk mengetahui (1) gambaran komitmen berpacaran, (2) gambaran kualitas persahabatan, (3) hubungan antara komitmen berpacaran dengan kualitas persahabatan pada remaja akhir di Universitas Pendidikan Indonesia (UPI). Responden pada penelitian ini ialah mahasiswa di UPI yang berada pada usia remaja akhir yang memiliki pasangan (pacar) dan juga teman dekat (sahabat). Terdapat 200 orang responden sesuai kriteria yang telah ditentukan dalam penelitian ini. Penelitian ini menggunakan metode kuantitatif dan menggunakan teknik purposive sampling. Pengumpulan data dilakukan melalui penyebaran kuesioner dengan memanfaatkan aplikasi google docs. Kuesioner berisikan skala komitmen berpacaran yang peneliti adaptasi dan modifikasi dari Commitment Measurenment milik Rusbult (1983) dan skala kualitas persahabatan yang juga peneliti adaptasi dan modifikasi dari Friendship Quality Scale (FQS) milik Bukowski, Hoza, dan Bovin (Ponti, dkk, 2010). Data yang diperoleh dianalisis menggunakan teknik korelasi Spearman Rho. Hasil penelitian menunjukkan (1) remaja akhir di UPI memiliki komitmen berpacaran yang cenderung rendah yaitu sebanyak 41% dari 200 orang responden, (2) remaja akhir di UPI memiliki kualitas persahabatan yang cenderung rendah yaitu sebanyak 36,5% dari 200 orang responden, dan (3) terdapat hubungan yang antara komitmen berpacaran dengan kualitas persahabatan pada remaja akhir di Universitas Pendidikan Indonesia.---------This study aims to describe (1) the image of commitment dating (2) the big picture of quality of friendship, and (3) relationship between Commitment Dating with the quality of friendship late adolescence at Indonesia University of Education (UPI). Responders of this study are late adolescence at Indonesia University of Education who are in commitment relationship and having close friends. There are around 200 people with the appropriate criteria have been determined in this study. This study uses quantitative methods and using purposive sampling technique. Data collected through questionnaires by using google docs app. The questionnaire contains of a commitment dating scale that is adapted by researcher and modification of Commitment Measurenment owned by Rusbult (1983) and so is Quality Scale (FQS) belongs to Bukowski, Hoza, and Bovin (Ponti et al, 2010). Data was analyzed by using Spearman Rho correlation techniques. The result showed that (1) late adolescence of UPI who are in committed relationship that tends to lower as many as 41% of the 200 respondents, (2) late adolescence of UPI have a rather low quality friendships as many as 36.5% of the 200 respondents, and (3) there is a relationship between commitment dating with the quality of friendship in late teens at UPI

New CDK8 inhibitors as potential anti-leukemic agents – Design, synthesisand biological evaluation

Cyclin-dependent kinase 8 (CDK8) plays a vital role in regulating cell transcription either through its association with the mediator complex or by the phosphorylation of transcription factors. CDK8-mediated activation of oncogenes has proved to be important in a variety of cancer types including hematological malignancies. We have designed and synthesized a series of new synthetic steroids. The compounds were evaluated as CDK8 inhibitors in vitro. The three most potent compounds exhibit Kd-values towards CDK8 in the low nanomolar range (3.5–18 nM). Furthermore, the compounds displayed selectivity for CDK8 in a panel of 465 different kinases. The cell studies indicated a selectivity to kill AML-cancer cell lines compared to normal cell lines.publishedVersionUnit Licence Agreemen

A single extracellular amino acid in Free Fatty Acid Receptor 2 defines antagonist species selectivity and G protein selection bias

Free Fatty Acid Receptor 2 is a GPCR activated by short chain fatty acids produced in high levels in the lower gut by microbial fermentation of non-digestible carbohydrates. A major challenge in studying this receptor is that the mouse ortholog does not have significant affinity for antagonists that are able to block the human receptor. Docking of exemplar antagonists from two chemical series to homology models of both human and mouse Free Fatty Acid Receptor 2 suggested that a single lysine - arginine variation at the extracellular face of the receptor might provide the basis for antagonist selectivity and mutational swap studies confirmed this hypothesis. Extending these studies to agonist function indicated that although the lysine - arginine variation between human and mouse orthologs had limited effect on G protein-mediated signal transduction, removal of positive charge from this residue produced a signalling-biased variant of Free Fatty Acid Receptor 2 in which Gi-mediated signalling by both short chain fatty acids and synthetic agonists was maintained whilst there was marked loss of agonist potency for signalling via Gq/11 and G12/13 G proteins. A single residue at the extracellular face of the receptor thus plays key roles in both agonist and antagonist function

Embryonic exposure to produced water can cause cardiac toxicity and deformations in Atlantic cod (Gadus morhua) and haddock (Melanogrammus aeglefinus) larvae

Regular discharges of produced water from the oil and gas industry represents the largest direct discharge of effluent into the marine environment worldwide. Organic compound classes typically reported in produced water include saturated hydrocarbons, monoaromatic and polyaromatic hydrocarbons (MAHs, PAHs) as well as oxygenated compounds, such as phenols, acids and ketones. This forms a cocktail of known and suspect toxicants, but limited knowledge is yet available on the sub-lethal toxicity of produced water to cold-water marine fish species. In the present work, we conducted a 4-day exposure of embryos of Atlantic cod (Gadus morhua) and haddock (Melanogrammus aeglefinus) to produced water extracts equivalent to 1:50, 1:500 and 1:5000 times dilutions of raw effluent. No significant reduction in survival or hatching success was observed, however, for cod, hatching was initiated earlier for exposed embryos in a concentration-dependent manner. During recovery, significantly reduced embryonic heart rate was observed for both species. After hatch, larvae subjected to embryonic exposure to produced water extracts were smaller, and displayed signs of cardiotoxicity, jaw and craniofacial deformations. In order to improve risk assessment and regulation of produced water discharges, it is important to identify which produced water components contribute to these effects.acceptedVersio

Cardiorespiratory fitness and future risk of venous thromboembolism

This is the peer reviewed version of the following article: Evensen, L. H., Isaksen, T., Brækkan, S. K. & Hansen, J.-B. (2019). Cardiorespiratory fitness and future risk of venous thromboembolism. Journal of Thrombosis and Haemostasis, 17(12), 2160-2168., which has been published in final form at https://doi.org/10.1111/jth.14619. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.Background - Cardiorespiratory fitness (CRF) is a strong predictor of future arterial cardiovascular disease and premature mortality. However, there are limited data on the association between CRF and the risk of incident venous thromboembolism (VTE). Objectives - To investigate whether estimated CRF (eCRF) was associated with the risk of incident VTE in a cohort recruited from the general population. Methods - Participants (n = 10 393) from the sixth survey of the Tromsø Study (2007—08) were included, and incident VTEs were recorded up to 31 December 2016. CRF was estimated in sex‐specific algorithms based on age, waist circumference, resting heart rate, and self‐reported physical activity. Hazard ratios (HRs) with 95% confidence intervals (CIs) of VTE according to categories of eCRF were estimated in Cox regression models adjusted for sex with age as timescale. The impact of weight status was evaluated in analyses stratified by weight category. Results - There were 176 incident VTEs during follow‐up. Compared with individuals with eCRF 100% of age‐predicted had 46% (HR 0.54; 95% CI 0.39‐0.77) and 67% (HR 0.33; 95% CI 0.20‐0.54) lower VTE risk, respectively. Compared with overweight/obese individuals with eCRF Conclusions - Higher eCRF was associated with lower risk of incident VTE. The association was independent of weight categories, suggesting that higher eCRF may modify the association between obesity and VTE

The role of goal-setting in urban climate governance

This article argues that goal-setting is an important, albeit understudied, part of urban climate governance scholarship. By using goal-setting theory, the article introduces concepts and perspectives capable of shedding new light on the political aspect of cities' climate strategic work. Climate goal-setting is studied within a wider urban governance context, as a way to activate a multitude of internal and external actors for shared goals and purposes. The article analyses levels of ambiguities of urban climate goals, and in light of different politico-institutional settings it explores possible contextual implications for cities’ climate governance. Through a comparative analysis of four cities – Copenhagen, Cape Town, Oslo and Gothenburg, the article identifies two distinct approaches. An inclusive approach containing ambiguous all-encompassing climate goals, consensus-oriented political decision-making, a broad administrative entity with weak mandate and close and long-term stakeholder collaboration. An efficiency-oriented approach including clear and problem focused climate goals, instrumental political decision-making, a special-purpose administrative entity with a wide and clear mandate and targeted and temporary stakeholder collaboration. The article concludes by posing some key questions that should guide further research on the exact relationship between these variables

Biodegradation-mediated alterations in acute toxicity of water-accommodated fraction and single crude oil components in cold seawater

Hydrocarbon biodegradation may be slower in cold Arctic than in temperate seawater, and this will affect the toxicity time window of the hydrocarbons. In this study, the acute toxicities of water-soluble phases of 1,3-dimethylnaphthalene, phenanthrene, fluoranthene, and low energy water-accommodated fractions (LE-WAFs) of an evaporated (200 °C+) crude oil, were screened by a Microtox bioassay during biodegradation in cold seawater (4–5 °C). The water-solubility of fluoranthene was too low to provoke a toxic response at any time, whereas the toxicity of 1,3-dimethylnaphthalene and phenanthrene decreased over time in relation to biotransformation of these compounds. In LE-WAFs, the Microtox EC50 was associated with biodegradation of the predominant hydrocarbons (naphthalenes, 2- to 3-ring PAH), as well as with phenol degradation products. The acute toxicities of single hydrocarbons and LE-WAFs persisted for a longer period in the cold seawater than previously shown at higher seawater temperatures. These results suggest implications for fate and effects assessment of hydrocarbons after oil spills in cold environments, like the Arctic. However, further biodegradation studies using Arctic seawater and relevant species for toxicity testing are needed for confirmation.acceptedVersio